Jill P Buyon, M.D.

A Novel Role of Endothelin-1 in Linking Toll-like Receptor 7-mediated Inflammation to Fibrosis in Congenital Heart Block
Alvarez, David; Briassouli, Paraskevi; Clancy, Robert M; Zavadil, Jiri; Reed, Joanne H; Abellar, Rosanna G; Halushka, Marc; Fox-Talbot, Karen; Barrat, Franck J; Buyon, Jill P
2011 Sep 2;286(35):30444-30454, Journal of biological chemistry
Autoimmune associated congenital heart block (CHB) may result from pathogenic cross-talk between inflammatory and profibrosing pathways. Incubation of macrophages with immune complexes (IC) composed of Ro60, a target of the pathologic maternal autoantibodies necessary for CHB, hY3 ssRNA, and affinity-purified anti-Ro60 antibody induces the Toll-like receptor 7 (TLR7)-dependent generation of supernatants that provoke a fibrosing phenotype in human fetal cardiac fibroblasts. We show herein that these cells are a major source of TGFbeta and that endothelin-1 (ET-1) is one of the key components responsible for the profibrosing effects generated by stimulated macrophages. Supernatants from macrophages incubated with IC induced the fibroblast secretion of TGFbeta, which was inhibited by treating the macrophages with an antagonist of TLR7. Under the same conditions, the induced fibroblast secretion of TGFbeta was decreased by inhibitors of the ET-1 receptors ETa or ETb or by an anti-ET-1 antibody but not by an isotype control. Exogenous ET-1 induced a profibrosing phenotype, whereas fibroblasts transfected with either ETa or ETb siRNA were unresponsive to the profibrosing effects of the IC-generated macrophage supernatants. Immunohistochemistry of the hearts from two fetuses dying with CHB revealed the presence of ET-1-producing mononuclear cells in the septal region in areas of calcification and fibrosis. In conclusion, these data support a novel role of ET-1 in linking TLR7 inflammatory signaling to subsequent fibrosis and provide new insight in considering therapeutics for CHB
— id: 136998, year: 2011, vol: 286, page: 30444, stat: Journal Article,

Binding of Anti-SSA Antibodies to Apoptotic Fetal Cardiocytes Stimulates Urokinase Plasminogen Activator (uPA)/uPA Receptor-Dependent Activation of TGF-beta and Potentiates Fibrosis
Briassouli, Paraskevi; Rifkin, Daniel; Clancy, Robert M; Buyon, Jill P
2011 Nov 15;187(10):5392-5401, Journal of immunology
In congenital heart block (CHB), binding of maternal anti-SSA/Ro Abs to fetal apoptotic cardiocytes impairs their removal by healthy cardiocytes and increases urokinase plasminogen activator (uPA)/uPA receptor (uPAR)-dependent plasmin activation. Because the uPA/uPAR system plays a role in TGF-beta activation, we evaluated whether anti-Ro binding to apoptotic cardiocytes enhances plasmin-mediated activation of TGF-beta, thereby promoting a profibrosing phenotype. Supernatants from cocultures of healthy cardiocytes and apoptotic cardiocytes bound by IgG from a mother whose child had CHB (apoptotic-CHB-IgG [apo-CHB-IgG]) exhibited significantly increased levels of active TGF-beta compared with supernatants from cocultures of healthy cardiocytes and apoptotic cardiocytes preincubated with IgG from a healthy donor. Treatment of the culture medium with anti-TGF-beta Ab or TGF-beta inhibitor (SB431542) abrogated the luciferase response, thereby confirming TGF-beta dependency. Increased uPA levels and activity were present in supernatants generated from cocultures of healthy cardiocytes and apo-CHB-IgG cardiocytes compared with healthy cardiocytes and apoptotic cardiocytes preincubated with IgG from a healthy donor, respectively. Treatment of apo-CHB-IgG cardiocytes with anti-uPAR or anti-uPA Abs or plasmin inhibitor aprotinin prior to coculturing with healthy cardiocytes attenuated TGF-beta activation. Supernatants derived from cocultures of healthy cardiocytes and apo-CHB-IgG cardiocytes promoted Smad2 phosphorylation and fibroblast transdifferentiation, as evidenced by increased smooth muscle actin and collagen expression, which decreased when fibroblasts were treated with supernatants from cocultures pretreated with uPAR Abs. These data suggested that binding of anti-Ro Abs to apoptotic cardiocytes triggers TGF-beta activation, by virtue of increasing uPAR-dependent uPA activity, thus initiating and amplifying a cascade of events that promotes myofibroblast transdifferentiation and scar
— id: 140530, year: 2011, vol: 187, page: 5392, stat: Journal Article,

Pregnancy outcomes in patients with autoimmune diseases and anti-Ro/SSA antibodies
Brucato, Antonio; Cimaz, Rolando; Caporali, Roberto; Ramoni, Veronique; Buyon, Jill
2011 Feb;40(1):27-41, Clinical reviews in allergy & immunology
Anti-Ro/SSA antibodies are associated with neonatal lupus (congenital heart block (CHB), neonatal transient skin rash, hematological and hepatic abnormalities), but do not negatively affects other gestational outcomes, and the general outcome of these pregnancies is now good, when followed by experienced multidisciplinary teams. The prevalence of CHB, defined as an atrioventricular block diagnosed in utero, at birth, or within the neonatal period (0-27 days after birth), in the offspring of an anti-Ro/SSA-positive women is 1-2%, of neonatal lupus rash around 10-20%, while laboratory abnormalities in asymptomatic babies can be detected in up to 27% of cases. The risk of recurrence of CHB is ten times higher. Most of the mothers are asymptomatic at delivery and are identified only by the birth of an affected child. Half of these asymptomatic women develop symptoms of a rheumatic disease, most commonly arthralgias and xerophtalmia, but few develop lupus nephritis. A standard therapy for CHB is still matter of investigation, although fluorinated corticosteroids have been reported to be effective for associated cardiomyopathy. Serial echocardiograms and obstetric sonograms, performed at least every 1-2 weeks starting from the 16th week of gestational age, are recommended in anti-Ro/SSA-positive pregnant women to detect early fetal abnormalities that might be a target of preventive therapy
— id: 120804, year: 2011, vol: 40, page: 27, stat: Journal Article,

Autoantibodies in systemic lupus erythematosus: comparison of historical and current assessment of seropositivity
Ippolito, A; Wallace, D J; Gladman, D; Fortin, P R; Urowitz, M; Werth, V; Costner, M; Gordon, C; Alarcon, G S; Ramsey-Goldman, R; Maddison, P; Clarke, A; Bernatsky, S; Manzi, S; Bae, S-C; Merrill, J T; Ginzler, E; Hanly, J G; Nived, O; Sturfelt, G; Sanchez-Guerrero, J; Bruce, I; Aranow, C; Isenberg, D; Zoma, A; Magder, L S; Buyon, J; Kalunian, K; Dooley, M A; Steinsson, K; van Vollenhoven, R F; Stoll, T; Weisman, M; Petri, M
2011 ;20(3):250-255, Lupus
Systemic lupus erythematosus (SLE) is characterized by multiple autoantibodies and complement activation. Recent studies have suggested that anti-nuclear antibody (ANA) positivity may disappear over time in some SLE patients. Anti-double-stranded DNA (dsDNA) antibody titers and complement levels may vary with time and immunosuppressive treatment, while the behavior of anti-extractable nuclear antigen (ENA) over time is less well understood. This study sought to determine the correlation between historical autoantibody tests and current testing in patients with SLE. Three hundred and two SLE patients from the ACR Reclassification of SLE (AROSE) database with both historical and current laboratory data were selected for analysis. The historical laboratory data were compared with the current autoantibody tests done at the reference laboratory and tested for agreement using percent agreement and Kappa statistic. Serologic tests included ANA, anti-dsDNA, anti-Smith, anti-ribonucleoprotein (RNP), anti-Ro, anti-La, rheumatoid factor (RF), C3 and C4. Among those historically negative for immunologic markers, a current assessment of the markers by the reference laboratory generally yielded a low percentage of additional positives (3-13%). However, 6/11 (55%) of those historically negative for ANA were positive by the reference laboratory, and the reference laboratory test also identified 20% more patients with anti-RNP and 18% more with RF. Among those historically positive for immunologic markers, the reference laboratory results were generally positive on the same laboratory test (range 57% to 97%). However, among those with a history of low C3 or C4, the current reference laboratory results indicated low C3 or C4 a low percentage of the time (18% and 39%, respectively). ANA positivity remained positive over time, in contrast to previous studies. Anti-Ro, La, RNP, Smith and anti-dsDNA antibodies had substantial agreement over time, while complement had less agreement. This variation could partially be explained by variability of the historical assays, which were done by local laboratories over varying periods of time. Variation in the results for complement, however, is more likely to be explained by response to treatment. These findings deserve consideration in the context of diagnosis and enrolment in clinical trials
— id: 134136, year: 2011, vol: 20, page: 250, stat: Journal Article,

Maternal and Fetal Factors Associated With Mortality and Morbidity in a Multi-Racial/Ethnic Registry of Anti-SSA/Ro-Associated Cardiac Neonatal Lupus
Izmirly, Peter M; Saxena, Amit; Kim, Mimi Y; Wang, Dan; Sahl, Sara K; Llanos, Carolina; Friedman, Deborah; Buyon, Jill P
2011 Nov 1;124(18):1927-1935, Circulation
Background- Cardiac manifestations of neonatal lupus include conduction disease and, rarely, an isolated cardiomyopathy. This study was initiated to determine the mortality and morbidity of cardiac neonatal lupus and associated risk factors in a multi-racial/ethnic US-based registry to provide insights into the pathogenesis of antibody-mediated injury and data for counseling. Methods and Results- Three hundred twenty-five offspring exposed to maternal anti-SSA/Ro antibodies with cardiac neonatal lupus met entry criteria. Maternal, fetal echocardiographic, and neonatal risk factors were assessed for association with mortality. Fifty-seven (17.5%) died, 30% in utero. The probability of in utero death was 6%. The cumulative probability of survival at 10 years for a child born alive was 86%. Fetal echocardiographic risk factors associated with increased mortality in a multivariable analysis of all cases included hydrops and endocardial fibroelastosis. Significant predictors of in utero death were hydrops and earlier diagnosis, and of postnatal death were hydrops, endocardial fibroelastosis, and lower ventricular rate. Isolated heart block was associated with a 7.8% case fatality rate, whereas the concomitant presence of dilated cardiomyopathy or endocardial fibroelastosis quadrupled the case fatality rate. There was a significantly higher case fatality rate in minorities compared with whites, who were at a lower risk of hydrops and endocardial fibroelastosis. Pacing was required in 70%; cardiac transplantation was required in 4 children. Conclusion- Nearly one fifth of fetuses who develop cardiac neonatal lupus die of complications predicted by echocardiographic abnormalities consistent with antibody-associated disease beyond the atrioventricular node. The disparity in outcomes observed between minorities and whites warrants further investigation
— id: 140527, year: 2011, vol: 124, page: 1927, stat: Journal Article,

{beta}2-Glycoprotein I and Protection from Anti-SSA/Ro60-Associated Cardiac Manifestations of Neonatal Lupus
Reed, Joanne H; Clancy, Robert M; Purcell, Anthony W; Kim, Mimi Y; Gordon, Tom P; Buyon, Jill P
2011 Jul 1;187(1):520-526, Journal of immunology
One mechanism to molecularly explain the strong association of maternal anti-Ro60 Abs with cardiac disease in neonatal lupus (NL) is that these Abs initiate injury by binding to apoptotic cardiomyocytes in the fetal heart. Previous studies have demonstrated that beta(2)-glycoprotein I (beta(2)GPI) interacts with Ro60 on the surface of apoptotic Jurkat cells and prevents binding of anti-Ro60 IgG. Accordingly, the current study was initiated to test two complementary hypotheses, as follows: 1) competition between beta(2)GPI and maternal anti-Ro60 Abs for binding apoptotic induced surface-translocated Ro60 occurs on human fetal cardiomyocytes; and 2) circulating levels of beta(2)GPI influence injury in anti-Ro60-exposed fetuses. Initial flow cytometry experiments conducted on apoptotic human fetal cardiomyocytes demonstrated dose-dependent binding of beta(2)GPI. In competitive inhibition experiments, beta(2)GPI prevented opsonization of apoptotic cardiomyocytes by maternal anti-Ro60 IgG. ELISA was used to quantify beta(2)GPI in umbilical cord blood from 97 neonates exposed to anti-Ro60 Abs, 53 with cardiac NL and 44 with no cardiac disease. beta(2)GPI levels were significantly lower in neonates with cardiac NL. Plasmin-mediated cleavage of beta(2)GPI prevented binding to Ro60 and promoted the formation of pathogenic anti-Ro60 IgG-apoptotic cardiomyocyte complexes. In aggregate these data suggest that intact beta(2)GPI in the fetal circulation may be a novel cardioprotective factor in anti-Ro60-exposed pregnancies
— id: 134451, year: 2011, vol: 187, page: 520, stat: Journal Article,

Association of the idiotype:Antiidiotype antibody ratio with the efficacy of intravenous immunoglobulin treatment for the prevention of recurrent autoimmune-associated congenital heart block
Routsias J.G.; Kyriakidis N.C.; Friedman D.M.; Llanos C.; Clancy R.; Moutsopoulos H.M.; Buyon J.; Tzioufas A.G.
2011 ;63(9):2783-2789, Arthritis & rheumatism
Objective Congenital heart block (CHB), a manifestation of neonatal lupus, is associated with maternal anti-Ro/SSA and anti-La/SSB autoantibodies and recurs in ~18% of subsequent pregnancies. This study was undertaken to investigate the effect of the idiotype:antiidiotype (Id:anti-Id) antibody ratio in the ability of intravenous immunoglobulin (IVIG) administered during subsequent pregnancies to prevent CHB. Methods We studied 16 anti-Ro/SSA and anti-La/SSB-positive pregnant women from the Preventive IVIG Therapy for Congenital Heart Block study who had previously given birth to a child with neonatal lupus. In 3 of the mothers, the study pregnancy resulted in the birth of a child with neonatal lupus (2 with CHB and 1 with rash). Sequential serum samples were obtained from all mothers immediately before the administration of IVIG during pregnancy and were evaluated for antibodies against the major B cell epitope 349-364aa of La/SSB (idiotype) and its antiidiotypic antibodies. Results Following IVIG treatment, serum titers of anti-La(349-364) (Id antibodies) decreased in 80% of the mothers, and in 60% an increase in anti-Id antibodies against anti-La(349-364) was observed. The Id:anti-Id ratio was significantly higher in mothers whose offspring developed neonatal lupus compared to mothers who gave birth to a healthy child (P < 0.0001). Removal of anti-Id antibodies substantially increased the reactivity against La(349-364) in sera from 5 of 7 mothers tested. All IVIG preparations were examined for Id and anti-Id antibody activity. IVIG from batches administered to mothers who gave birth to a healthy child had an Id:anti-Id activity ratio of <1, in contrast to that given to mothers who gave birth to a child with neonatal lupus. Addition of the IVIG preparations to the maternal sera further enhanced antiidiotypic activity (by up to 4.7-fold) in 11 of 13 patients studied. Conclusion This is the first study in humans to demonstrate that IVIG influences the Id-anti-Id network of a specific pathogenic autoantibody. Specifically, we showed that IVIG enhanced the anti-Id antibody response in pregnant women with anti-La/SSB antibodies. A high Id:anti-Id ratio in both the IVIG preparation and the maternal serum may explain the absence of an effect of IVIG in preventing recurrent neonatal lupus in some cases.
— id: 137457, year: 2011, vol: 63, page: 2783, stat: Journal Article,

International consensus for a definition of disease flare in lupus
Ruperto N; Hanrahan L; Alarcon G; Belmont H; Brey R; Brunetta P; Buyon J; Costner M; Cronin M; Dooley M; Filocamo G; Fiorentino D; Fortin P; Franks A Jr; Gilkeson G; Ginzler E; Gordon C; Grossman J; Hahn B; Isenberg D; Kalunian K; Petri M; Sammaritano L; Sanchez-Guerrero J; Sontheimer R; Strand V; Urowitz M; von Feldt J; Werth V; Merrill J
2011 ;20(5):453-462, Lupus
The Lupus Foundation of America (LFA) convened an international working group to obtain a consensus definition of disease flare in lupus. With help from the Paediatric Rheumatology International Trials Organization (PRINTO), two web-based Delphi surveys of physicians were conducted. Subsequently, the LFA held a second consensus conference followed by a third Delphi survey to reach a community-wide agreement for flare definition. Sixty-nine of the 120 (57.5%) polled physicians responded to the first survey. Fifty-nine of the responses were available to draft 12 preliminary statements, which were circulated in the second survey. Eighty-seven of 118 (74%) physicians completed the second survey, with an agreement of 70% for 9/12 (75%) statements. During the second conference, three alternative flare definitions were consolidated and sent back to the international community. One hundred and sixteen of 146 (79.5%) responded, with agreement by 71/116 (61%) for the following definition: 'A flare is a measurable increase in disease activity in one or more organ systems involving new or worse clinical signs and symptoms and/or laboratory measurements. It must be considered clinically significant by the assessor and usually there would be at least consideration of a change or an increase in treatment.' The LFA proposes this definition for lupus flare on the basis of its high face validity
— id: 134268, year: 2011, vol: 20, page: 453, stat: Journal Article,

Doppler fetal mechanical PR interval prolongation with positive maternal anti-RNP but negative SSA/Ro and SSB/La auto-antibodies
Acherman, Ruben J; Friedman, Deborah M; Buyon, Jill P; Schwartz, Joel; Castillo, William J; Rollins, Robert C; Evans, William N
2010 Aug;30(8):797-799, Prenatal diagnosis
— id: 114634, year: 2010, vol: 30, page: 797, stat: Journal Article,

A novel role of endothelin-1 in linking Ro60-ssRNA immune complexes to cardiac fibrosis in congenital heart block
Alvarez-Carbonell D.; Zavadil J.; Abellar R.; Barrat F.; Clancy R.; Buyon J.
2010 ;62:2231-2231, Arthritis & rheumatism
Passively acquired anti-Ro associated congenital heart block (CHB) likely results from pathologic crosstalk between inflammatory and fibrosing pathways eventuating in scarring. The target, Ro60, complexed with uridine rich ssRNA induces TLR7-dependent macrophage production of supernatants capable of trans-differentiating human fetal cardiac fibroblasts. This study addressed the molecular components responsible for the TLR-dependent profibrosing effects. In seeking the link between an inflammatory response of macrophages and the profibrotic effect on fibroblasts, a microarray analysis comparing the mRNA expression profile of macrophages stimulated with hY3 (ssRNA associated with Ro60) or immune complexes consisting of Ro60-hY3-IgG fraction containing anti-Ro60 antibodies (IC) in the presence or absence of IRS661 (antagonist of TLR7) was performed. Gene expression of the vasoconstrictor endothelin-1 (ET-1), recently shown to promote dermal fibrosis, was significantly upregulated by ~ 4 fold and confirmed by RT-PCR. Incubation of macrophages with either hY3 or IC increased secretion of ET-1 from 2.64 pg/mL (baseline) to 9.67 pg/mL (p <.0001) and 7.52 pg/mL (p =.0003) respectively. Pre-treatment with IRS661 decreased hY3-induced secretion to 3.83 pg/mL and IC to 3.98 pg/mL. The direct effect of ET-1 (100 nM) on these fibroblasts (shown to express both ETa and ETb receptors) resulted in a profibrosing phenotype as demonstrated by a) TGFbeta secretion (13 pg/mL vs. 772 pg/mL, p =.03), b) increased collagen release (18 ng/mL baseline vs. 772 ng/mL, p =.05, and c) expression of a-smooth muscle actin (alpha-smac). ET-1-induced TGFbeta secretion was significantly decreased (p =.03) by each of the following: BQ-123, an ETa antagonist (119 pg/mL), BQ-788, an ETb antagonist (145 pg/mL), and anti-ET-1 antibody (211 pg/mL), but not isotype control antibody (691 pg/mL). Similarly, ET-1-induced collagen secretion was significantly decreased (p =.05) by BQ-123 (147 ng/mL collagen), BQ-788 (247 ng/mL), anti-ET-1 antibody (253 ng/mL), and anti-TGFbeta antibody (239 ng/ml), but not isotype control (815 ng/mL). Predictably, pretreatment of fibroblasts with either BQ-123, BQ-788 or incubation of supernatants with anti-ET-1 antibody, but not isotype control, significantly inhibited the profibrosing readouts (increased TGFbeta, collagen and alpha-smac) induced by supernatants from macrophages stimulated with either hY3 or IC. Additionally, fibroblasts transfected with either ETa or ETb siRNA, but not scrambled siRNA, also significantly inhibited the profibrosing readouts induced stimulated macrophages. Immuno-histochemistry of a heart from a fetus dying with CHB revealed ET-1/2/3 antibody staining but not isotype IgG control in the septal region in areas showing calcification, fibrosis, and mononuclear cell infiltrates (not seen in an age-matched fetal heart). In conclusion, these data suggest that macrophage secretion of ET-1 may be one mechanism linking TLR inflammatory signaling to subsequent fibrosis and provide new insight in considering therapeutics for CHB
— id: 130921, year: 2010, vol: 62, page: 2231, stat: Journal Article,

Discordance between self-report and physician-assessed disease activity in patients with systemic lupus erythematosus (SLE): Implications for clinical trial design and clinical care
Askanase A.D.; Castrejon I.; Buyon J.P.; Yazici Y.; Pincus T.
2010 ;62:1857-1857, Arthritis & rheumatism
Purpose: To analyze agreement levels between patient (PT) and physician (MD) assessments in 50 patients with SLE seen in usual care, including a) global PT and MD estimates of status; b) patient self-report scores on the SLAQ (Systemic Lupus Activity Questionnaire) and MDHAQ (Multidimensional Health Assessment Questionnaire) for physical function (FN), pain (PN), patient global estimate (PTGL), fatigue (FT), RAPID3 (FN, PN, and PTGL), and review of systems checklist (SX); c) physician-scored indices SLEDAI-2K (SLE Disease Activity Index), BILAG (British Isles Lupus Assessment Group index), SLAM (SLE Activity Measure) and ECLAM (European Consensus Lupus Activity Measurement). Methods: A cross-sectional study was performed in 50 consecutive SLE patients of one rheumatologist. Patients completed the SLAQ and MDHAQ, including PTGL. The rheumatologist scored a physician global estimate (MDGL) (scored 0-3 in 0.1 increments) without knowledge of PTGL, and completed the SLEDAI 2K, BILAG, SLAM, and ECLAM. Agreement levels of various measures were analyzed using Spearman rank order correlations. Results: The study included 45 women and 5 men, mean age 38.7 years, mean disease duration 7.3 years, 36% Caucasian, 18% Black, 26% Hispanic, 18% Asian. The mean MDGL (1.10+/-0.62), PTGL (3.11 +/-2.81) and SLE indices (SLEDAI 5.02+/-3.75; BILAG 4.60+/-4.31; SLAM 3.86+/-2.92; ECLAM 1.97+/-1.37) indicated mild/moderate lupus activity. The correlation between MDGL and PTGL of rho=0.14 was not statistically significant. Correlations between MDGL and SLE indices were significant, rho=0.60-0.72 (p<0.001). Correlations between PTGL and patient measures also were significant, rho=0.58-0.87 (p<0.001). However, PTGL was correlated at lower levels with SLE indices - significantly with BILAG and SLAM (0.35-0.40; p<0.01), and not significantly with SLEDAI or ECLAM. MDGL was not correlated significantly with any patient measure or index. (Table Presented) Conclusion: MDGL and PTGL are not correlated significantly, an observation made previously. MDGL was correlated significantly with all physician-derived indices, and PTGL was correlated significantly with all patient-derived measures and indices. By contrast, MDGL was correlated at much lower, nonsignificant levels with patient-derived measures and indices, and PTGL was correlated at lower levels with physician-derived indices. Further analysis of these discordances may clarify the clinical relevance of various measures in clinical trials, and may lead to improved care and compliance in patients with SLE
— id: 130931, year: 2010, vol: 62, page: 1857, stat: Journal Article,

Frequency of neuro-psychiatric dysfunction in anti-SSA/SSB exposed children with and without neonatal lupus
Askanase, A D; Izmirly, P M; Katholi, M; Mumtaz, J; Buyon, J P
2010 ;19(3):300-306, Lupus
Neonatal lupus is a model of passively acquired autoimmunity whereby anti-SSA/Ro-SSB/La antibodies target the fetal heart and neonatal skin in a minority of cases. Since neuro-psychiatric impairment has been reported in humans and mice exposed prenatally to a variety of maternal autoantibodies including anti-Ro/La, this study was initiated to evaluate the potential neurotoxic effects of these specific autoantibodies and the overall frequency of autoimmune diseases, general health, and somatic growth of children with neonatal lupus and their unaffected siblings. In addition to the general health questionnaires maintained on family members enrolled in the Research Registry for Neonatal Lupus (RRNL), specific questionnaires related to neuro-psychiatric development were sent to all mothers whose children (both affected and unaffected) were older than 5 years of age. Controls consisted of healthy friends. Of 121 anti-Ro exposed children meeting the inclusion criteria, information was returned on 104 (33 cardiac manifestations of neonatal lupus, 20 rash, and 51 unaffected siblings) and 22 of the friend controls. The mean age of all of the children was 14.5 years (range 5-39). In total, 42 (40%) of the 104 anti-Ro exposed children were reported to have a neuro-psychiatric disorder, compared with 6 (27%) of the friend controls (p = 0.34). For 8 (24%) of the congenital heart block (CHB) children (6 boys, 2 girls) the mothers reported attention problems. Four, all boys, were on stimulants. Of the rash children, 4 (20%) (2 boys, 2 girls) had attention problems with one boy on Ritalin. Of the unaffected siblings, 9 (18%) (8 boys and 1 girl) had attention problems with 3 boys on stimulants. One (5%) of the control children (a girl) had attention problems, not requiring therapy. There was no statistical difference in attention problems between the groups (p = 0.120). Behavioral problems were present in all groups with no statistical differences noted. The prevalence of depression, anxiety, developmental delays, learning, hearing, and speech problems were not significantly different between groups. In the CHB children, one boy has nephrotic syndrome and one girl has psoriasis. In the rash children, one girl has juvenile rheumatoid arthritis. In the unaffected group there are five children with autoimmune diseases, two with inflammatory bowel diseases (one boy and one girl), one boy has a spondyloarthropathy, one girl has alopecia areata and one young woman has Antiphospholipid syndrome. In the control group one boy has Henoch Schonlein purpura. There were four cases of hypothyroidism, possibly secondary to Hashimoto's thyroiditis, three in boys with CHB and one in a girl with rash. None of the unaffected siblings or controls had hypothyroidism. Parental reporting of neuro-psychiatric abnormalities was high in anti-Ro exposed children regardless of the neonatal lupus manifestation. However, medication use was limited and although the frequency of this reporting was greater than friend controls, it did not reach significance
— id: 107768, year: 2010, vol: 19, page: 300, stat: Journal Article,

Binding of apoptotic fetal cardiocytes by anti-Ro/La antibodies stimulates uPA/uPAR-dependent activation of TGFbeta and potentiates fibrosis
Briassouli P.; Rifkin D.; Buyon J.P.; Clancy R.M.
2010 ;62:489-489, Arthritis & rheumatism
Purpose: Organ injury induced by antibodies characteristic of Sjogren Syndrome and Systemic Lupus Erythematosus, while varied in the adult and fetus, may share in common a link between apoptosis and ultimate fibrosis. In congenital heart block (CHB), binding of maternal anti-Ro/La antibodies to apoptotic cardiocytes impairs their removal by healthy cardiocytes and increases uPA/uPAR-dependent plasmin activation. Immunological staining of CHB hearts reveals AV node TGFb staining and TGFb activation promotes fibroblast trans-differentiation, a scarring phenotype. Since the uPA/uPAR system plays a role in TGFb activation, this study evaluated whether anti-Ro/La binding to apoptotic cardiocytes via plasmin activation stimulates TGFb and promotes a profibrosing phenotype. Methods and Results: Initial analysis showed increased TGFb in supernatants from co-cultures of healthy cards and apoptotic cards incubated with IgG fractions from mothers whose sera contain anti-Ro/La antibodies and who had a child with CHB (apo-CHB-IgG) compared to co-cultures of healthy cards and apoptotic cards incubated with control IgG (apo-nl-IgG). Using a luciferase bioassay of active TGFb, supernatants from co-cultures of healthy cards and apo-CHB-IgG cards exhibited increased levels of active TGFb compared to those cocultured with apo-nl-IgG cards (511pg/ml CHB-IgG RLU vs 217 nl-IgG RLU; p=0.007; n=7). Abrogation of RLU via anti-TGFb antibody or TGFb inhibitor (SB431542) confirmed TGFb activation was solely due to TGFb. Significantly increased uPA levels (903 pg/ml vs 508 pg/ml; p =0.01; n=5) and uPA activity (0.8 units vs 0.04 units; p=0.005; n=3) were demonstrated in supernatants generated from coculture of healthy cards and apo CHB-IgG cards compared to healthy cards and apo nl-IgG, respectively. To determine whether uPA activity was responsible for TGFb activation, coculture experiments were conducted in which the apo-CHB-IgG cards were treated with anti-uPAR or anti-uPA antibodies or the plasmin inhibitor aprotinin prior to coculturing with healthy cards. In all instances treatments attenuated TGFb activation and uPA activity. To evaluate the profibrotic role of the observed TGFb activation, supernatants from either apo-CHB-IgG or apo-nl-IgG cocultures with healthy cards were applied to serum deprived cardiac fibroblasts. Only supernatants derived from cocultures of healthy cards and apo-CHB-IgG cardiocytes promoted transdifferentiation as evidenced by increased SMAc staining, an effect that was decreased when fibroblasts were treated with supernatants where cocultures were pretreated with uPAR antibodies. Supporting the hypothesis that increased uPA activity is causally related to the pathogenesis of CHB, cord blood samples from 12 of 18 children with CHB exhibited increased uPA activity and uPAR cleavage compared to 4 of 14 non CHB children exposed to maternal anti-Ro antibodies. Conclusions: These data suggest that binding of anti-Ro/La antibodies to apoptotic cardiocytes by virtue of increased uPAR-dependent uPA activity trigger TGFb activation thus initiating and amplifying a cascade of events that promote myofibroblast transdifferentiation and scar
— id: 130926, year: 2010, vol: 62, page: 489, stat: Journal Article,

Role of the Urokinase Plasminogen Activator Receptor in Mediating Impaired Efferocytosis of Anti-SSA/Ro-Bound Apoptotic Cardiocytes: Implications in the Pathogenesis of Congenital Heart Block
Briassouli, Paraskevi; Komissarova, Elena V; Clancy, Robert M; Buyon, Jill P
2010 Aug 6;107(3):374-387, Circulation research
Rationale: Binding of maternal anti-Ro/La antibodies to cognate antigen expressed on apoptotic cardiocytes decreases clearance by healthy cardiocytes, which may contribute to the development of autoimmune associated congenital heart block and fatal cardiomyopathy. Objective: Given recent evidence implicating the urokinase plasminogen activator receptor (uPAR) as a 'don't eat me' signal during efferocytosis, experiments addressed whether surface bound anti-Ro antibodies inhibit apoptotic cell removal via an effect on the expression/function of the urokinase-type plasminogen activator protease uPA/uPAR system. Methods and Results: As assessed by flow cytometry and confocal microscopy, uPAR colocalizes and interacts with Ro60 on the surface of apoptotic human fetal cardiocytes. Blocking of uPAR enhances phagocytosis of apoptotic cardiocytes by healthy cardiocytes and reverses the anti-Ro60-dependent impaired clearance of apoptotic cardiocytes. Binding of anti-Ro60 antibodies to apoptotic cardiocytes results in increased uPAR expression, as well as enhanced uPA activity. The binding of anti-Ro60 did not alter other surface molecules involved in cell recognition (calreticulin, CD31, or CD47). Conclusions: These data suggest that increased uPAR expression and uPA activity induced by anti-Ro60 binding to the apoptotic fetal cardiocyte provide a molecular basis by which these antibodies inhibit efferocytosis and ultimately lead to scar of the fetal conduction system and working myocardium
— id: 111585, year: 2010, vol: 107, page: 374, stat: Journal Article,

Passively acquired anti-SSA/Ro antibodies are required for congenital heart block following ovodonation but maternal genes are not
Brucato, Antonio; Ramoni, Veronique; Penco, Silvana; Sala, Elena; Buyon, Jill; Clancy, Robert
2010 Oct;62(10):3119-3121, Arthritis & rheumatism
Anti-SSA/Ro antibodies are necessary but not sufficient to provoke autoimmune-associated congenital heart block (CHB). Genetic factors are likely contributory. Accordingly, HLA-related candidates and single-nucleotide polymorphisms in the promoter region of tumor necrosis factor alpha and codon 10 in transforming growth factor beta1 (TGFbeta1) were evaluated in a unique family: the surrogate mother (anti-SSA/Ro positive), the biologic father, and the CHB-affected child (product of ovodonation). There was an HLA mismatch between the affected child and the surrogate mother. However, both the biologic and the surrogate mothers shared DQ2 and the profibrosing leucine polymorphism at codon 10 of TGFbeta. In conclusion, we observed that CHB can develop in a genetically unrelated child exposed in utero to anti-SSA/Ro antibodies. Testing for anti-SSA/Ro antibodies might be considered in women undergoing artificial fertilization. It is possible that there is no direct association of maternal genes beyond a contributory role in generating the autoantibody
— id: 120803, year: 2010, vol: 62, page: 3119, stat: Journal Article,

Genome-wide association study of cardiac manifestations of neonatal lupus identifies risk variants in the ERG, TCF19,C6orf10 and MICB-TNF-AIF1 region
Clancy R.; Marion M.C.; Kaufman K.M.; Ramos P.S.; Adler A.; Harley J.B.; Langefeld C.D.; Buyon J.P.
2010 ;19:21-21, Lupus
Objectives: Isolated congenital heart block (CHB) is highly associated with maternal anti-Ro/SSA antibodies. CHB carries a substantial mortality, approaching 30%, and morbidity, with over 60% of surviving affected children requiring lifelong pacemakers. To date, complete atrioventricular (AV) block is irreversible. This is a rare disease and factors beyond requisite maternal autoantibody are unknown. Data from twin studies and the 10-fold increased recurrence rate of cardiac neonatal lupus (NL) in subsequent pregnancies indicate a strong genetic contribution to risk. We posit that fetal genes influence the response to maternal autoantibodies. Methods: Children of European ancestry (n=116) with cardiac NL were identified from the U.S. Research Registry for Neonatal Lupus. Cases were genotyped using the Illumina 370K SNP platform and merged with 3351 controls from the International Consortium on Systemic Lupus Erythematosus Genetics (SLEGEN). Standard quality control and admixture-adjusted tests of association were computed. Results: Outside the HLA region, a strong association was detected at 21q22, upstream from the transcriptional regulator ERG (rs743446, p=5.45E-06, OR = 2.40). Within the HLA, associated regions include PSORS1C1 (rs3130544, p = 1.94E-07, OR = 2.77) and a missense mutation (proline to serine) at TCF19 (rs7750641, p = 1.58E-07, OR = 2.79), at Class I; several variants in the MICB-NFKBIL1- LTA-TNF-LTB-AIF1 region at Class III (rs2230365, p= 1.00E-03, OR=0.46; rs2857595, p= 1.96E-09, OR = 2.37; rs3128982, p= 6.40E-06, OR=1.86; and rs3099844, p= 4.52E-10, OR= 3.34; and the C6orf10 locus at class II (rs3115553, p=2.69E-05, OR=1.81; rs6457536, pa=1.74E-05, OR=1.84; and rs7775397 (p = 1.35E-09, OR=3.30). These are consistent with our previous results (Clancy 2002). With the exception of the HLA region, no loci previously implicated in autoimmune diseases achieved genome-wide significance in the CHB children. Conclusion: These results suggest that genetic variation near ERG, PSORS1C1, LTA/TNF/LTB and C6orf10 in the fetus may promote an abnormal tissue response initiated by exposure to maternal autoantibodies. Identification of risk loci is an incremental step towards discovery of a fetal genetic component that contributes to the anti-SSA/Ro associated development of life-long cardiac damage
— id: 134746, year: 2010, vol: 19, page: 21, stat: Journal Article,

Ro60-Associated Single-Stranded RNA Links Inflammation with Fetal Cardiac Fibrosis via Ligation of TLRs: A Novel Pathway to Autoimmune-Associated Heart Block
Clancy, Robert M; Alvarez, David; Komissarova, Elena; Barrat, Franck J; Swartz, Jordan; Buyon, Jill P
2010 Feb 15;184(4):2148-2155, Journal of immunology
Activation of TLR by ssRNA after FcgammaR-mediated phagocytosis of immune complexes (IC) may be relevant in autoimmune-associated congenital heart block (CHB) where the obligate factor is a maternal anti-SSA/Ro Ab and the fetal factors, protein/RNA on an apoptotic cardiocyte and infiltrating macrophages. This study addressed the hypothesis that Ro60-associated ssRNAs link macrophage activation to fibrosis via TLR engagement. Both macrophage transfection with noncoding ssRNA that bind Ro60 and an IC generated by incubation of Ro60-ssRNA with an IgG fraction from a CHB mother or affinity purified anti-Ro60 significantly increased TNF-alpha secretion, an effect not observed using control RNAs or normal IgG. Dependence on TLR was supported by the significant inhibition of TNF-alpha release by IRS661 and chloroquine. The requirement for FcgammaRIIIa-mediated delivery was provided by inhibition with an anti-CD16a Ab. Fibrosis markers were noticeably increased in fetal cardiac fibroblasts after incubation with supernatants generated from macrophages transfected with ssRNA or incubated with the IC. Supernatants generated from macrophages with ssRNA in the presence of IRS661 or chloroquine did not cause fibrosis. In a CHB heart, but not a healthy heart, TLR7 immunostaining was localized to a region near the atrioventricular groove at a site enriched in mononuclear cells and fibrosis. These data support a novel injury model in CHB, whereby endogenous ligand, Ro60-associated ssRNA, forges a nexus between TLR ligation and fibrosis instigated by binding of anti-Ro Abs to the target protein likely accessible via apoptosis
— id: 106500, year: 2010, vol: 184, page: 2148, stat: Journal Article,

Identification of candidate loci at 6p21 and 21q22 in a genome-wide association study of cardiac manifestations of neonatal lupus
Clancy, Robert M; Marion, Miranda C; Kaufman, Kenneth M; Ramos, Paula S; Adler, Adam; Harley, John B; Langefeld, Carl D; Buyon, Jill P
2010 Nov;62(11):3415-3424, Arthritis & rheumatism
OBJECTIVE: Cardiac manifestations of neonatal lupus, comprising atrioventricular conduction defects and cardiomyopathy, occur in fetuses exposed to anti-Ro/SSA antibodies, and carry substantial mortality. There is strong evidence of a genetic contribution to the risk. This study was undertaken to evaluate single-nucleotide polymorphisms (SNPs) for associations with cardiac neonatal lupus. METHODS: Children of European ancestry with cardiac neonatal lupus (n = 116) were genotyped using the Illumina 370K SNP platform and merged with 3,351 controls. Odds ratios (ORs) and 95% confidence intervals (95% CIs) for association with cardiac neonatal lupus were determined. RESULTS: The 17 most significant associations with cardiac neonatal lupus were found in the HLA region. The region near the MICB gene showed the strongest variant (rs3099844; P(dom) = 4.52 x 10(-10) , OR 3.34 [95% CI 2.29-4.89]), followed by a missense variant within C6orf10 (rs7775397; P(dom) = 1.35 x 10(-9) , OR 3.30), which lies between NOTCH4 and BTNL2, and several SNPs near the tumor necrosis factor alpha gene, including rs2857595 (P(add) = 1.96 x 10(-9) , OR 2.37), rs2230365 (P(add) = 1.00 x 10(-3) , OR 0.46), and rs3128982 (P(add) = 6.40 x 10(-6) , OR 1.86). Outside the HLA region, an association was detected at 21q22, upstream of the transcription regulator ets-related isoform 1 (rs743446; P = 5.45 x 10(-6) , OR 2.40). HLA notwithstanding, no individual locus previously implicated in autoimmune diseases achieved genome-wide significance. CONCLUSION: These results suggest that variation near genes related to inflammatory and apoptotic responses may promote cardiac injury initiated by passively acquired autoantibodies
— id: 114171, year: 2010, vol: 62, page: 3415, stat: Journal Article,

Evaluation of fetuses in a study of intravenous immunoglobulin as preventive therapy for congenital heart block: Results of a multicenter, prospective, open-label clinical trial
Friedman, Deborah M; Llanos, Carolina; Izmirly, Peter M; Brock, Brigit; Byron, John; Copel, Joshua; Cummiskey, Karen; Dooley, Mary Anne; Foley, Jill; Graves, Cornelia; Hendershott, Colleen; Kates, Richard; Komissarova, Elena V; Miller, Michelle; Pare, Emmanuelle; Phoon, Colin K L; Prosen, Tracy; Reisner, Dale; Ruderman, Eric; Samuels, Philip; Yu, Jerry K; Kim, Mimi Y; Buyon, Jill P
2010 Apr;62(4):1138-1146, Arthritis & rheumatism
OBJECTIVE: The recurrence rate of anti-SSA/Ro-associated congenital heart block (CHB) is 17%. Sustained reversal of third-degree block has never been achieved. Based on potential reduction of maternal autoantibody titers as well as fetal inflammatory responses, intravenous immunoglobulin (IVIG) was evaluated as preventive therapy for CHB. METHODS: A multicenter, prospective, open-label study based on Simon's 2-stage optimal design was initiated. Enrollment criteria included the presence of anti-SSA/Ro antibodies in the mother, birth of a previous child with CHB/neonatal lupus rash, current treatment with < or = 20 mg/day of prednisone, and <12 weeks pregnant. IVIG (400 mg/kg) was given every 3 weeks from week 12 to week 24 of gestation. The primary outcome was the development of second-degree or third-degree CHB. RESULTS: Twenty mothers completed the IVIG protocol before the predetermined stopping rule of 3 cases of advanced CHB in the study was reached. CHB was detected at 19, 20, and 25 weeks; none of the cases occurred following the finding of an abnormal PR interval on fetal Doppler monitoring. One of these mothers had 2 previous children with CHB. One child without CHB developed a transient rash consistent with neonatal lupus. Sixteen children had no manifestations of neonatal lupus at birth. No significant changes in maternal titers of antibody to SSA/Ro, SSB/La, or Ro 52 kd were detected over the course of therapy or at delivery. There were no safety issues. CONCLUSION: This study establishes the safety of IVIG and the feasibility of recruiting pregnant women who have previously had a child with CHB. However, IVIG at low doses consistent with replacement does not prevent the recurrence of CHB or reduce maternal antibody titers
— id: 111773, year: 2010, vol: 62, page: 1138, stat: Journal Article,

Dysregulation of the microvascular as assessed by expression of protective and injury associated markers is reflected in the non-lesional non-sunexposed skin of patients with lupus nephritis
Izmirly P.M.; Meehan S.; Xu S.X.; Askanase A.D.; Merrill J.T.; Buyon J.P.; Clancy R.M.
2010 ;62:1190-1190, Arthritis & rheumatism
Purpose: Coagulation is one of the first pathways to be elicited by vascular injury, and its activation is followed by proinflammatory phenomena, in part due to loss of the anti-inflammatory activity of both the Protein C pathway and membrane Endothelial Protein C receptor (mEPCR). It has been recently demonstrated that mEPCR is highly expressed in the cortical peritubular capillaries of kidneys from patients (pts) with active lupus nephritis compared to normal human kidney. Profound upregulation of mEPCR was observed even in areas absent tubulointerstitial damage. This study addressed the hypothesis that changes in the microvasculature extend beyond the clinically targeted organ and that dysregulation is a fundamental characteristic of SLE. Methods: The study included SLE pts in whom renal disease was considered active as assessed by proteinuria and urinary sediment. Renal biopsies were performed in all pts. Thirty skin biopsies from non-lesional nonsunexposed skin (buttocks) were obtained in 26 pts (23 females, 3 males) and five healthy controls (4 females, 1 male). The paraffin skin sections were individually stained with specific antibodies against mEPCR and adiponectin (protective markers), ICAM-1 (proinflammatory) and CD31 (pan endothelial marker). Immunohistochemistry (IHC) was scored by counting peroxidase-brown labeled blood vessels (10-20 microns in diameter) without knowledge of the clinical information associated with the biopsy. The number of blood vessels with an intensity of at least 1+ were quantitatively scored with ranges 1-12. To account for the number of blood vessels per slide, the CD31 count had to be 12 to be included in the analysis. Results: The 28 renal biopsies comprised the following ISN/RPS classifications: 4 Class III, 7 Class IV, 8 Class V, I Class VI, 3 Class III/V, 3 Class IV/V. Nineteen percent of the pts had a GFR <60 (mean GFR, 82 ml/min). Abnormal laboratory values for complement and anti-dsDNA antibodies were reported in 72% and 75% of pts, respectively. Nephrotic range proteinuria was present in 37%. For IHC skin assessments of the controls, the mean score for mEPCR was 1 (highest 2), ICAM-1 was 4 (highest 7) and adiponectin was 1 (highest 2). In 17/25 (68%) of the SLE non-lesional non-sun exposed skin sections, mEPCR was expressed above the highest control. In 16/30 (53%) ICAM-1 staining exceeded 7. In contrast, only 6/25 (19%) expressed adiponectin above 2. For each specific stain there were no apparent differences between biopsy class, degree of proteinuria, presence of anti dsDNA or low complement levels. However, pts with mEPCR staining above 2 had higher GFR measurements than those with staining < 2 (88 ml/min +/- 31 versus 53 +/- 32, p= 0.0168). In contrast, GFR was unrelated to ICAM-1 and adiponectin expression. Conclusion: These data are consistent with the notion that there is widespread activation of the microvasculature. The capacity of endothelial cells to utilize anticoagulation pathways is not restricted to the kidney and expression of mEPCR in the microcirculation likely represents an attempt to limit microvascular inflammation in kidney and skin
— id: 130932, year: 2010, vol: 62, page: 1190, stat: Journal Article,

Mortality/morbidity in cardiac neonatal lupus and associated maternal/fetal risk factors
Izmirly P.M.; Saxena A.; Smith Z.; Buyon J.P.
2010 ;62:1438-1438, Arthritis & rheumatism
Purpose: The classic cardiac manifestations of neonatal lupus (cardiac-NL) include a spectrum of conduction dysfunction (1^st, 2^nd, or 3^rd degree heart block (CHB)) and more rarely cardiomyopathy which can be absent any conduction disease. This study was undertaken to update the mortality data of cardiac-NL in a large US based cohort and identify associated risk factors to further understand the pathogenesis of anti-SSA/Ro mediated injury and provide evidence based data for counseling women with these antibodies. Methods: Three hundred and one children with cardiac-NL (295 with CHB and 6 with isolated cardiomyopathy) enrolled in the Research Registry for Neonatal Lupus (RRNL) had sufficient medical records for review. The RRNL database was analyzed for the following potential mortality maternal risk factors: age at pregnancy, race/ethnicity, anti-SSB/La antibody status, health status and fetal risk factors: time of diagnosis, exposure to maternal non-fluorinated and fluorinated steroids during pregnancy, method of delivery, and gender. In addition, morbidity was assessed by the frequency of pacemaker placement and cardiac transplant. Results: Follow up of the children ranged from in utero death to adulthood. Of the 301 children with cardiac-NL, 53 (17.6%) died. Thirty percent died in utero or at birth, 41% died prior to six months of postnatal life and the remaining 29% died after 6 months. Mortality was higher among children born to non-Caucasian mothers compared to Caucasian mothers (33% vs 15% p=.0003). Maternal age was equivalent between the groups (29.1 dead vs 29.7 live). The maternal presence of anti-SSB/La antibodies was 74% for those whose children died and 63% for those whose children survived, which was not significant. A maternal diagnosis of Sjorgen's Syndrome and/or Systemic Lupus Erythematosus was not significantly associated with cardiac-NL death (53% in death vs 44%) suggesting that prior knowledge of maternal antibody status did not influence mortality. With regard to fetal factors, 86% of those who died were diagnosed with cardiac-NL during pregnancy compared to 85% who survived. For those diagnosed during pregnancy there was a trend towards early gestational diagnosis for those children that died compared to those that survived (21 vs 23.5 weeks p=.09). There was also a trend toward higher exposure to maternal fluorinated steroids after the diagnosis in children that died (53% vs 40% p=.09), however there was no difference in maternal use of non-fluorinated steroids in those that died vs those that survived (19% vs 16%). Most fetuses were delivered by C-section and this was not significantly associated with death (70% dead vs 75% live). Female gender was also not associated with outcome (49% who died were female vs 52% live). Eighty-five percent of children received a pacemaker, 43% within 9 days of birth, 20% between 9 days and one year. Five children (2%) received a heart transplant. Conclusion: Based on data from this large cohort, 17.6% of children born with cardiac-NL die from complications of the disease. Eighty-five percent required pacing and two percent required cardiac transplantation. Mortality was more prevalent in children born to non Caucasian mothers
— id: 130944, year: 2010, vol: 62, page: 1438, stat: Journal Article,

Evaluation of the risk of anti-SSA/Ro-SSB/La antibody-associated cardiac manifestations of neonatal lupus in fetuses of mothers with systemic lupus erythematosus exposed to hydroxychloroquine
Izmirly, Peter M; Kim, Mimi Y; Llanos, Carolina; Le, Phuong U; Guerra, Marta M; Askanase, Anca D; Salmon, Jane E; Buyon, Jill P
2010 Oct;69(10):1827-1830, Annals of rheumatic diseases
BACKGROUND: Based on the potential involvement of Toll-like receptor (TLR) signalling in the pathogenesis of neonatal lupus (NL), it was hypothesised that fetal exposure to hydroxychloroquine (HCQ), a TLR inhibitor, might reduce the risk of anti-SSA/Ro/SSB/La antibody-associated cardiac manifestations of NL (cardiac-NL). METHODS: Cardiac-NL children (N=50) and controls (N=151) were drawn from the following overlapping pregnancy studies: Research Registry for NL; PR Interval and Dexamethasone Evaluation in Cardiac-NL; and Predictors of Pregnancy Outcomes: Biomarkers in Antiphospholipid Syndrome and Systemic Lupus Erythematosus (SLE). Pregnancies met the following inclusion criteria: documentation of maternal anti-SSA/Ro/SSB/La antibodies at pregnancy, confirmation of medication use and child's outcome, a diagnosis of SLE before pregnancy and birth by 31 December 2007. RESULTS: Seven (14%) of the cardiac-NL children were exposed to HCQ compared with 56 (37%) of the controls (p=0.002; OR 0.28; 95% CI 0.12 to 0.63). Cases and controls were similar with respect to demographic and antibody status. Multivariable analysis adjusting for birth year, maternal race/ethnicity, antibody status, non-fluorinated steroid use and prior cardiac-NL risk yielded an OR associated with HCQ use of 0.46 (95% CI 0.18 to 1.18; p=0.10). CONCLUSION: This case-control study suggests that, in mothers with SLE with anti-SSA/Ro/SSB/La antibodies, exposure to HCQ during pregnancy may decrease the risk of fetal development of cardiac-NL. Prospective studies are needed for confirmation
— id: 138123, year: 2010, vol: 69, page: 1827, stat: Journal Article,

Cutaneous manifestations of neonatal lupus and risk of subsequent congenital heart block
Izmirly, Peter M; Llanos, Carolina; Lee, Lela A; Askanase, Anca; Kim, Mimi Y; Buyon, Jill P
2010 Apr;62(4):1153-1157, Arthritis & rheumatism
OBJECTIVE: Cutaneous disease associated with placental transport of maternal anti-SSA/Ro or anti-SSB/La antibodies is transient, and children often appear to be otherwise healthy. However, the impact of this manifestation of neonatal lupus (NL) on the risk of cardiac disease occurring in a future pregnancy is critical for family counseling and for powering preventive trials. The purpose of this study was to determine the recurrence rates of NL, with specific focus on cardiac NL following cutaneous NL in a child enrolled in the Research Registry for Neonatal Lupus (RRNL). METHODS: Fifty-eight families who were enrolled in the RRNL met the following inclusion criteria for our study: maternal anti-SSA/Ro or anti-SSB/La antibodies, a child with cutaneous NL, and a pregnancy subsequent to the child with cutaneous NL. RESULTS: The majority of the 58 mothers (78%) were Caucasian. Of 77 pregnancies that occurred following the birth of a child with cutaneous NL, the overall recurrence rate for any manifestation of NL was 49% (95% confidence interval [95% CI] 37-62%); 14 pregnancies (18.2%) were complicated by cardiac NL, 23 (29.9%) by cutaneous NL, and 1 (1.3%) by hematologic/hepatic NL. A subset analysis was restricted to the 39 children who were born after the initial child with cutaneous NL had been enrolled in the RRNL. The overall recurrence rate for NL was 36% (95% CI 20-52%); 5 pregnancies (12.8%) were complicated by cardiac NL and 9 (23.1%) by cutaneous NL. There were no significant differences in the following maternal risk factors for having a subsequent child with cardiac or cutaneous NL: age, race/ethnicity, anti-SSB/La status, diagnosis, use of nonfluorinated steroids, or breastfeeding. The sex of the subsequent fetus did not influence the development of cardiac or cutaneous NL. CONCLUSION: Based on data from this large cohort, the identification of cutaneous NL in an anti-SSA/Ro antibody-exposed infant is particularly important, since it predicts a 6-10-fold risk of a subsequent child developing cardiac NL
— id: 109039, year: 2010, vol: 62, page: 1153, stat: Journal Article,

Congenital heart block: identification of autoantibody binding site on the extracellular loop (domain I, S5-S6) of alpha(1D) L-type Ca channel
Karnabi, Eddy; Qu, Yongxia; Wadgaonkar, Raj; Mancarella, Salvatore; Yue, Yuankun; Chahine, Mohamed; Clancy, Robert M; Buyon, Jill P; Boutjdir, Mohamed
2010 Mar;34(2):80-86, Journal of autoimmunity
Congenital heart block (CHB) is an autoimmune disease associated with autoantibodies against intracellular ribonucleoproteins SSB/La and SSA/Ro. The hallmark of CHB is complete atrioventricular block. We have recently established that anti-SSA/Ro -SSB/La autoantibodies inhibit alpha(1D) L-type Ca current, I(Ca-L), and cross-react with the alpha(1D) Ca channel protein. This study aims at identifying the possible binding sites on alpha(1D) protein for autoantibodies from sera of mothers with CHB children. GST fusion proteins of the extracellular regions between the transmembrane segments (S5-S6) of each of the four alpha(1D) Ca channel protein domains I-IV were prepared and tested for reactivity with sera from mothers with CHB children and controls using ELISA. Sera containing anti-Ro/La autoantibodies from 118 mothers with CHB children and from 15 mothers with anti-Ro/La autoantibodies but have healthy children, and from 28 healthy mothers without anti-Ro/La autoantibodies and healthy children were evaluated. Seventeen of 118 (14.4%) sera from mothers with CHB children reacted with the extracellular loop of domain I S5-S6 region (E1). In contrast, only 2 of 28 (7%) of sera from healthy mothers (-anti-Ro/La) and healthy children reacted with E1 loop and none (0 of 15) of sera from healthy mothers (+anti-Ro/La) and healthy children reacted with the E1 loop. Preincubation of E1 loop with the positive sera decreased the O.D reading establishing the specificity of the response. Electrophysiological characterization of the ELISA positive sera and purified IgG showed inhibition (44.1% and 49.8%, respectively) of the alpha(1D) I(Ca-L) expressed in tsA201 cells. The inhibition was abolished when the sera were pre-incubated with E1 fusion protein. The results identified the extracellular loop of domain I S5-S6 of L-type Ca channel alpha(1D) subunit as a target for autoantibodies from a subset of mothers with CHB children. This novel finding provides insights into the potential development of therapeutic peptides that could bind to the pathogenic antibodies and prevent CHB
— id: 114633, year: 2010, vol: 34, page: 80, stat: Journal Article,

Anatomic and Pathologic Findings in Hearts from Fetuses and Infants with Cardiac Manifestations of Neonatal Lupus
Llanos, C; Clancy, RM; Abellar, R; Buyon, JP; Friedman, DM
2010 SEP ;72(3):270-271, Scandinavian journal of immunology
— id: 111795, year: 2010, vol: 72, page: 270, stat: Journal Article,

Flare assessment in systemic lupus erythematosus (SLE) patients treated with rituximab in the phase II/III EXPLORER trial
Merrill J.T.; Buyon J.P.; Furie R.; Latinis K.M.; Gordon C.; Hsieh H.-J.; Brunetta P.
2010 ;19:39-39, Lupus
Objectives: SLE patients (pts) with moderate-to-severe disease activity (>=1 BILAG A or >=2 BILAG B domain scores) despite background immunosuppressives and corticosteroids, were randomized to placebo (PLA) or rituximab (RTX). Although differences in primary and secondary outcome measures were not observed, an exploratory analysis was performed to evaluate the imObjectives: SLE patients (pts) with moderate-to-severe disease activity (>=1 BILAG A or >=2 BILAG B domain scores) despite background immunosuppressives and corticosteroids, were randomized to placebo (PLA) or rituximab (RTX). Although differences in primary and secondary outcome measures were not observed, an exploratory analysis was performed to evaluate the impact of RTX on disease flares. Our objectives were to assess in those pts who achieved response whether RTX affected: 1) time to moderate or severe flares, 2) annualized flare rates, and 3) prednisone usage during flares. Methods: Pts who achieved response (BILAG C, D, E scores for all domains before wk52) on monthly BILAG assessments were included in this analysis. Flares, defined as increased disease activity following achievement of response, were stratified as follows: Severe flare: >=1 BILAG A or >3 BILAG B domain scores; A flare: >=1 new BILAG A domain scores; Moderate flare: 2 BILAG B domain scores. Kaplan-Meier estimates were used to assess time-to-flare. Annualized flare rates were calculated using number of flares per patient with a Poisson regression model. Only flares that occurred after the protocol-mandated prednisone taper at 12 wks were included in the analysis of prednisone use during flares. Results: Responses were achieved in 58/88 (66.0%) PLA-treated and 127/169 (75.1%) RTX-treated pts during the study. No difference was found between rituximab and placebo in preventing or delaying moderate to severe flares. However, when BILAG A flares alone were examined, rituximab reduced the risk of an A flare after achieving a response by 52 weeks (hazard ratio=0.612; p=0.0524) and lowered the annualized A flare rate (0.86 +/- 1.47 (SD) vs 1.41 +/- 2.14; p=.038). Eighty-four of 169 (49.7%) rituximab-treated patients achieved low disease activity without subsequent A flares versus 31/88 (35.2%) patients in the placebo group (p=0.027). Prednisone rescue for A flares was similar in rituximab- (24%) and placebo-treated (14%) patients (p=.204). Conclusions: No conclusions about rituximab efficacy can be drawn from this post hoc analysis. This exploratory analysis suggests that assessment of BILAG A flares may distinguish potential treatment effects with more sensitivity than BILAG B flares, which capture modest changes in disease activity. If confirmed in other studies, this observation may help in the design of more robust clinical trial protocols
— id: 134745, year: 2010, vol: 19, page: 39, stat: Journal Article,

Rituximab: wanted dead or alive..
Merrill, Joan T; Buyon, Jill P
2010 Aug;62(8):2188-2191, Arthritis & rheumatism
— id: 112058, year: 2010, vol: 62, page: 2188, stat: Journal Article,

IRF5 SLE-risk haplotype is strongly enriched in anti-Ro positive mothers of Neonatal lupus patients with diverse diagnoses
Niewold T.B.; Kariuki S.N.; Franek B.S.; Buyon J.P.; Clancy R.M.
2010 ;62:495-495, Arthritis & rheumatism
Objective: In patients with systemic lupus erythematosus (SLE), IRF5 genotype is associated with anti-Ro autoantibodies. It is not clear whether this association with autoantibody profile is specific to SLE disease status. To examine this question, we assessed IRF5 genotypes in a large cohort of individuals who all had high titer anti-Ro autoantibodies and carried a variety of diagnoses including healthy/asymptomatic, Sjogren's syndrome (SS), and SLE. Methods: We studied 85 European-ancestry individuals recruited to the Research Registry for Neonatal Lupus who all had high titer anti-Ro autoantibodies and a child with neonatal lupus. The diagnoses of these subjects were as follows: 15 healthy/asymptomatic, 10 SLE, 19 SLE/SS, 20 SS, and 21 undifferentiated autoimmune disease (UAS). IRF5 SNPs were genotyped using Taqman primer-probe sets to define previously reported European-derived SLE risk and protective haplotypes. Results: The IRF5 SLE-risk haplotype defined by the rs10488631 C allele was enriched in subjects of all diagnoses except SS when compared to large published data sets of European-American controls [OR=2.30 (1.50-3.32), p=8.4x10^-5]. Each of the asymptomatic, SLE, SLE/SS, and UAS mothers showed a very similar increase in rs10488631 allele frequency when examined individually (ORs range from 2.00 to 3.15, no significant differences between the asymptomatic, SLE, SLE/SS and UAS groups). The rs10488631 C allele frequency in the subjects diagnosed with SS was essentially the same as controls (OR=1.05). Interestingly, the rs3807306 C allele was significantly increased in the SS patients as compared to subjects from the other 4 diagnostic categories [OR=2.34 (1.14-4.79), p=0.026]. Conclusions: There was a significant increase in the frequency of the IRF5 SLE-risk haplotype in subjects with anti-Ro antibodies with varying diagnoses which was of comparable magnitude to the anti-Ro/IRF5 association observed in SLE (OR<2). Interestingly, the SS cohort was distinct from the other neonatal lupus mothers with respect to IRF5 genotype, and a different SNP was associated with SS status. These data suggest that the genetic association of IRF5 with autoimmunity is strongly influenced by serologic profile, and that the anti-Ro association with IRF5 genotype extends beyond the SLE disease state to some degree
— id: 130941, year: 2010, vol: 62, page: 495, stat: Journal Article,

Clues To differentiate non-inflammatory from inflammatory symptoms in patients with Systemic Lupus erythematosus (SLE), using a multi-dimensional health assessment questionnaire (MDHAQ)
Pincus T.; Castrejon I.; Buyon J.P.; Tseng C.-E.; Izmirly P.M.; Yazici Y.; Askanase A.D.
2010 ;62:990-990, Arthritis & rheumatism
Purpose: To analyze whether quantitative scores on a multidimensional health assessment questionnaire (MDHAQ) provide clues to the likelihood of inflammatory versus non-inflammatory symptoms and concomitant fibromyalgia, an important challenge in clinical care, according to a global scale for noninflammatory symptoms completed by a rheumatologist in 50 patients with SLE seen in usual care. Methods: A cross-sectional study was performed in 50 consecutive SLE patients of one rheumatologist seen in usual care. On arrival at the clinic, patients completed a multidimensional health assessment questionnaire (MDHAQ) which includes scales for physical function (FN), 0-10 visual analog scales for pain (PN), global estimate (PTGL) and fatigue (FT), and a review of systems symptom checklist (SX). SLE patients also completed a self-report Systemic Lupus Assessment Questionnaire (SLAQ). The rheumatologist, unaware of MDHAQ and SLAQ scores, recorded a physician global estimate (MDGL) and an estimate of non-inflammatory symptoms, each scored on a 0-3 scale in 0.1 increments, as well as four SLE indices: SLEDAI-2K (SLE Disease Activity Index), BILAG (British Isles Lupus Assessment Group index), SLAM (SLE Activity Measure) with and without laboratory tests, and ECLAM (European Consensus Lupus Activity Measurement). SLE patients with scores of <0.5 on the noninflammatory symptom scale were regarded as low and those with scores >=0.5 high noninflammatory symptoms; the two groups were compared using the Mann-Whitney statistic. Results: The study included 45 women and 5 men, mean age 38.7 years, mean disease duration 7.3 years. Of the 50 patients, 16 had high and 34 low scores for non-inflammatory symptoms. Those with high scores for non-inflammatory symptoms had significantly higher scores for FN, PN, FT, PTGL, SX, SLAQ, and SLAM without laboratory tests, as well as significantly lower CRP. No significant differences were seen patients estimated as high and low scoring patients for SLEDAI, BILAG, SLAM, ECLAM, C3, C4, antiDsDNA, or ESR. Fewer than 50% of low patients had FN, PN, PTGL, or FT >=2, while 100% of high patients had FT >2, and 94% PTGL >2. All patients with high non-inflammatory symptoms (16/16) reported more than 5 SX, compared to 15/34 (44%) low patients, and 12/16 (75%) high patients reported >10 SX, compared to 6/34 (18%) low patients. (Table Presented) Conclusion: High scores for dysfunction, pain, fatigue, global estimates, and number of symptoms are common in SLE patients with high versus low levels of non-inflammatory symptoms. SLE indices do not distinguish between patients with high versus low levels of non-inflammatory symptoms. A simple global scale to estimate non-inflammatory symptoms may be informative in therapeutic decisions, particularly if consistent with patient questionnaire patterns
— id: 130928, year: 2010, vol: 62, page: 990, stat: Journal Article,

Association of plasma soluble E-selectin and adiponectin with carotid plaque in patients with systemic lupus erythematosus
Reynolds, Harmony R; Buyon, Jill; Kim, Mimi; Rivera, Tania L; Izmirly, Peter; Tunick, Paul; Clancy, Robert M
2010 Jun;210(2):569-574, Atherosclerosis
BACKGROUND: Systemic lupus erythematosus (SLE) is associated with premature atherosclerosis but the mechanisms underlying this association are not understood. The role of endothelial dysfunction is hypothesized. METHODS: In predominantly non-Caucasian patients with SLE (N=119) and controls (N=71), carotid ultrasonography was performed and circulating endothelial cells (CECs), soluble endothelial protein C receptor and gene polymorphism at A6936G, soluble E-selectin (sE-selectin), and adiponectin were assessed. RESULTS: Carotid plaque was more prevalent among patients than controls (43% vs 17%, p=0.0002). Mean CCA IMT was greater in patients compared to controls (0.59+/-0.19 mm vs 0.54+/-0.11 mm, p=0.03). Among SLE patients, plaque was not associated with smoking, body-mass index, LDL, triglycerides, homocysteine, C-reactive protein, anti-ds DNA antibody, C3, C4, SLE activity, or medications. Age and levels of soluble E-selectin and adiponectin were significantly higher in the SLE patients with plaque compared to those without plaque in univariate and multivariate analyses. sE-selectin and adiponectin were found to serve as independent predictors of carotid plaque and that elevations were persistent over more than one visit. Unexpectedly, these biomarkers were present despite clinical quiescence. CONCLUSION: Premature atherosclerosis is a consistent feature of SLE and extends across ethnicities. Higher levels of adiponectin may represent a physiological attempt to limit further endothelial damage already reflected by the elevation in sE-selectin and the observed increase in plaque represents overwhelming of this reparative process by atherogenic stimuli
— id: 109844, year: 2010, vol: 210, page: 569, stat: Journal Article,

IVIG Administration Enhances the Anti-Idiotypic Response to the Major Epitope Of La/SSB Autoantigen in Pregnant Women at High Risk for a Child with Chb
Routsias, JG; Kyriakidis, N; Friedman, D; Llanos, C; Clancy, R; Buyon, J; Tzioufas, AG
2010 SEP ;72(3):272-272, Scandinavian journal of immunology
— id: 111796, year: 2010, vol: 72, page: 272, stat: Journal Article,

Ro60 associated ssRNA links inflammation with fetal cardiac fibrosis via ligation of toll like receptors: A potential pathway to heart block
Alvarez D.; Komissarova E.V.; Swartz J.; BarraT F.; Rivera T.L.; Clancy R.M.; Buyon J.P.
2009 ;60:127-127, Arthritis & rheumatism
Purpose: Exploration of pathogenic mechanisms coupling anti-Ro/La antibodies to fetal cardiac injury has focused on the protein target of the maternal immune response. The relevance of Interferon Type I and Toll like receptor (TLR) signaling in SLE supports the potential importance of Ro associated ssRNAs in the cascade to congenital heart block (CHB). Objective: To address the hypothesis that ssRNA induces macrophage activation via TLR ligation following uptake of a complex of Ro60, hY3 or mutant pre 5S (m-pre5S) RNA and anti-Ro with release of proinflammatory and profibrotic factors which result in scarring of the conduction system and endocardial fibroelastosis. Method: In vitro conditions included evaluation of human macrophages for secretion of TNFa (inflammatory component) and transdifferentiation of human fetal cardiac fibroblasts and collagen production (fibrosing component) as well as staining of human fetal hearts. Results:The TLR component was evaluated by transfection of ssRNA. Treatment of IFNg-primed macrophages with hY3 or m-pre5S RNA (2.5mg each) significantly stimulated TNFa release (1,121+/-373pg/mL p=0.02, N=14, and 1,072+/-338pg/mL respectively vs resting macrophages 92+/-40 pg/mL, p=0.01, N=14), an effect not observed with transfected ssRNA41 (control RNA) or modified hY3 RNA (base substitution of hY3, A/U). Both theTLR7/8 antagonist IRS661(32ng/mL) and chloroquine (10mM) significantly decreased TNFa release induced by either hY3 or m-pre5S RNA (IRS661: 159+/-77pg/mL p=0.03, N=9 for hY3, and 71+/-29pg/mL p=0.03, N=9 for pre-5S; chloroquine: 267+/-89pg/mL p=0.03, N=9 for hY3, and 180+/-70pg/mL p=0.03, N=9 for pre-5S). Immune complexes generated by incubation of an IgG fraction from a CHB mother with native Ro60-hY3 (CHB IgG Ro60-hY3) significantly increased TNFa secretion compared to CHB IgG Ro60-ssRNA41 or normal IgG (healthy donor absent anti-Ro) incubated with Ro60-hY3 (687+/-248pg/mL vs 246+/-103pg/mL p=0.05, N=3 vs 18+/-11pg/mL p<0.001, N=3). Fibrosis was evaluated using the identical supernatants (sups). Transdifferentiation of fibroblasts (SMAc staining, N=5) was markedly increased by incubation with sups generated from macrophages + hY3 or m-pre5s RNA, CHB IgG Ro60-hY3, not control ssRNAs or normal IgG Ro60-hY3. IRS661 and chloroquine each abrogated induced SMAc. Collagen secretion was stimulated by sups of macs + hY3 (766+/-82ng/mL, p=0.003 vs macs) compared to ssRNA41 (150+/-30ng/mL, p=NS vs macs) or hY3-A/U (197+/-27ng/mL, p=NS vs macs), and macs + CHB IgG Ro60-hY3 (650ng/mL) compared to normal IgG Ro60-hY3 (200ng/mL). TLR7 expressing mononuclear cells were observed in a CHB heart, not normal heart, and localized near the AV groove at a site enriched in fibrosis. Conclusion: These data support a novel injury model in CHB whereby endogenous ligand, Ro60 associated ssRNA, forges a nexus between TLR ligation and fibrosis instigated by binding of anti-Ro to the target protein accessible via apoptosis
— id: 130343, year: 2009, vol: 60, page: 127, stat: Journal Article,

Urinary cytology evaluation in SLE: Implications for identification of active nephritis and scoring renal domains on activity indices
Askanase A.D.; Thomas D.B.; Chowdhury A.; Purcell J.; Buyon J.P.
2009 ;60:906-906, Arthritis & rheumatism
Purpose: Perhaps with the exclusion of the elusive RBC cast, there is no single laboratory test that can unambiguously predict the presence of active renal disease in patients with SLE. Evaluation of urinary cytology is a valuable tool in identifying early transplant rejection with urinary lymphocytes (>20/10 high power field) preceding the rise in creatinine. This study addressed the value of urinary cytology as a marker of active renal disease in a consecutive sampling of SLE patients. Method: Seventy spot urine specimens were obtained from the first 50 patients fulfilling 4 ACR criteria seen in an outpatient setting. These were subjected to cytological evaluation in addition to protein/creatinine ratios. Results: Of the 50 patients, 19 have lupus nephritis defined by a kidney biopsy, 11 of which were active at the time of study as defined by a 24 hour protein excretion of >1 gram. As expected, the average protein/creatinine ratio on the spot urinalysis (UA) of patients with active nephritis compared to patients without active nephritis was 2.71 compared to.41. Clinical data on these patients further substantiated the presence of active renal disease; mean serum albumin (3.3 vs. 4.1), creatinine (1.36 vs 1.11), complement C3 (84 vs. 107), and C4 (16 vs. 21), and anti-dsDNA (228 vs. 52). Examination of the urinary cyto-diagnostic sediments (evaluated per 10 high power fields) revealed that in patients with active nephritis, there were significantly more acantocytes (dysmorphic RBCs), target, and isomorphic RBCs per high powered field compared to inactive patients 8/0, 27/1, 127/7 respectively. However, acantocytes, known to have the highest predictive value for glomerular injury, were only present in 2 patients with active nephritis, whereas all the others had none. The urinary neutrophils were higher in patients with active disease (235 vs. 63) but there was a high incidence of vaginal contamination (substantiated by the presence squamous epithelial cells in 9 of the 17 samples from active nephritis patients, and 33 of 53 inactive), After eliminating the contaminated samples, the PMNs were 131 vs 58 p=NS. The most striking difference between groups was in the number of lymphocytes, 81 vs 3 which remained significant even after removing the contaminated samples (92 vs 5, p=0.008). Of all the patients without active nephritis only 1 had urinary lymphocytes above 20 (21). Since contamination with vaginal fluid can have an impact on the presence of isomorphic RBCs we also examined the average number of RBCs after eliminating the contaminated samples, 62 vs. 12, p=NS. Conclusion: The presence of increased lymphocytes, not total WBC, on cytological urine exam suggests the diagnosis of active nephritis which may provide insight to activity vs chronicity, a critical clinical dilemma. The value of total RBCs and WBCs is diminished by contamination with vaginal fluid implying caution regarding scoring of renal domains
— id: 130349, year: 2009, vol: 60, page: 906, stat: Journal Article,

Use of pharmacogenetics, enzymatic phenotyping, and metabolite monitoring to guide treatment with azathioprine in patients with systemic lupus erythematosus
Askanase, Anca D; Wallace, Daniel J; Weisman, Michael H; Tseng, Chung-E; Bernstein, Lana; Belmont, H Michael; Seidman, Ernest; Ishimori, Mariko; Izmirly, Peter M; Buyon, Jill P
2009 Jan;36(1):89-95, Journal of rheumatology
OBJECTIVE: Individualized therapy based on genetic background and monitoring of metabolites can optimize drug safety and efficacy. Such an approach is available for azathioprine (AZA), the thiopurine antimetabolite. AZA exerts therapeutic effects when metabolized to the active thiopurine nucleotide, 6-thioguanine (6-TGN). In inflammatory bowel disease (IBD), 6-TGN levels in the target range of 235-400 pmol/8x10(8) red blood cells (RBC) are associated with a high likelihood of response. Our objective was to evaluate whether drug escalation based on metabolite levels improves efficacy and maintains safety in patients with systemic lupus erythematosus (SLE). METHODS: We conducted a 6-month open-label dose-escalation clinical study of patients with active SLE treated with azathioprine dosed by body weight and metabolite levels. The primary endpoint was >or=50% improvement in any one parameter of disease activity, or 50% decrease in glucocorticoid dose. RESULTS: Of 50 patients enrolled in the study, 21 achieved clinical responses, 13 of whom had 6-TGN<235 pmol/8 x10(8) RBC. Ten patients had no clinical response at 6 months, yet achieved either therapeutic IBD 6-TGN levels (>235, n=4) or received maximum AZA dose>or=3.5 mg/kg (n=6). In 19 patients the drug was discontinued prematurely due to side effects or SLE activity. For those patients in whom either liver function test or white blood cell count abnormalities were encountered, metabolites guided attribution to drug or disease activity. CONCLUSION: Clinical responses in SLE can occur at levels of 6-TGN lower than the target range established for IBD. During followup, measurements of AZA metabolites may provide a rational approach to safety
— id: 93731, year: 2009, vol: 36, page: 89, stat: Journal Article,

Congenital heart block (CHB) after ovodonation
Brucato, A; Clancy, RM; Ramoni, V; Borghi, A; Sadou, Y; Pisoni, MP; Buyon, JP
2009 AUG 8 ;139(31-32):15S-15S, Swiss medical weekly
— id: 101608, year: 2009, vol: 139, page: 15S, stat: Journal Article,

Autoimmune associated congenital heart block: integration of clinical and research clues in the management of the maternal / foetal dyad at risk
Buyon, J P; Clancy, R M; Friedman, D M
2009 Jun;265(6):653-662, Journal of internal medicine
One of the strongest associations with autoantibodies directed to components of the SSA/Ro-SSB/La ribonucleoprotein complex is the development of congenital heart block (CHB) in an offspring, an alarming prospect facing 2% of primigravid mothers with these reactivities. This risk is 10-fold higher in women who have had a previously affected child with CHB. Anti-Ro/La antibodies are necessary but insufficient to cause disease. In vitro and in vivo experiments suggest that the pathogenesis involves exaggerated apoptosis, macrophage/myfibroblast crosstalk, TGFbeta expression and extensive fibrosis in the conducting system and in some cases surrounding myocardium. A disturbing observation is the rapidity of disease progression, with advanced heart block and life-threatening cardiomyopathy observed <2 weeks from normal sinus rhythm. Once 3rd degree (complete) block is identified, reversal has never been achieved, despite dexamethasone. Current strategies include the evaluation of an early echocardiographic marker of injury, such as a prolonged PR interval and the use of IVIG as a preventative measure for pregnancies of mothers with previously affected children
— id: 99251, year: 2009, vol: 265, page: 653, stat: Journal Article,

Updates on lupus and pregnancy
Buyon, Jill P
2009 ;67(3):271-275, Bulletin of the NYU Hospital for Joint Diseases
This review focuses on events subsequent to planning a pregnancy and addresses three components of concern for women with systemic lupus erythematosus: maternal, placental, and fetal. Flare rates are generally low for patients who are clinically stable at conception. For patients who have never had renal disease, there is no frm evidence that they will develop active renal disease simply due to being pregnant. For patients who begin pregnancy with an abnormal creatinine (> 2 mg/dl is ill advised), risks include hypertension, preeclampsia, high rate of fetal loss, and possible further deterioration of renal function. Discontinuation of angiotensin converting enzyme inhibitors, angiotensin receptor blockers, and mycophenalate is mandatory. Elevated levels of sVEGF-1 may be a harbinger of preeclampsia. For patients with anti-phospholipid antibodies detected in the frst trimester of pregnancy, the lupus anticoagulant per se may be the strongest predictor of pregnancy complications. For women with anti-SSA/Ro antibodies the risk of having a child with congenital heart block is 2% which rises to a recurrence rate of 18%. Information on current approaches to prevention and treatment of heart complications of neo-natal lupus is provided
— id: 104897, year: 2009, vol: 67, page: 271, stat: Journal Article,

Cardiac manifestations of neonatal lupus erythematosus: guidelines to management, integrating clues from the bench and bedside
Buyon, Jill P; Clancy, Robert M; Friedman, Deborah M
2009 Mar;5(3):139-148, Nature clinical practice. Rheumatology
One of the strongest clinical associations with autoantibodies against components of the SSA/Ro-SSB/La ribonucleoprotein complex is the development of congenital heart block in an offspring, an alarming prospect facing 2% of primigravid mothers with these reactivities. This risk is increased tenfold in women who have had a previous child with congenital heart block. Accumulated evidence suggests that anti-SSA/Ro and anti-SSB/La antibodies are necessary but insufficient for fetal disease. Basic and clinical research is heavily focused on identifying fetal and environmental factors that convert disease susceptibility to disease development. A disturbing observation that has emerged from current research efforts is the rapidity of disease progression, with advanced heart block and life-threatening cardiomyopathy being observed less than 2 weeks after detection of a normal sinus rhythm. Once third-degree block is unequivocally identified, reversal has never been achieved, despite dexamethasone treatment. Accordingly, strategies aimed at preventing disease before irrevocable scarring ensues assume a high priority. One approach has been the implementation of serial echocardiography to monitor for a prolonged PR interval. Intravenous immunoglobulin is being evaluated as a potential prophylactic approach in mothers who have previously had an affected child
— id: 94462, year: 2009, vol: 5, page: 139, stat: Journal Article,

Towards genetic correlates of cardiac manifestations of neonatal lupus: A genome-wide association study
Clancy R.M.; Marion M.C.; Kaufman K.M.; Adler A.; Harley J.B.; Langefeld C.D.; Buyon J.P.
2009 ;60:1230-1230, Arthritis & rheumatism
Purpose: Maternal anti-Ro antibodies are highly associated with heart block and/or cardiomyopathy (cardiac neonatal lupus (NL) in an offspring. Monozygotic twin studies and the 10 fold increased recurrence rate of cardiac NL in a subsequent pregnancy implicate a strong genetic influence on risk of disease. Here, we report the first genome-wide association study of cardiac NL. Methods: Caucasian children (n=117) with cardiac NL were identified from an extensive collection of DNA in the U.S. Research Registry for Neonatal Lupus (RRNL). Two criteria were required: 1) cardiac NL defined as heart block (1st, 2nd,3rd, degree) documented by electrocardiogram (if 1st degree), echocardiogram, history of pacemaker, or statement in the medical record; and/or presence of cardiac injury which included autopsy evidence of a mononuclear infiltrate in the endocardium, myocardium and pericardium and/or endocardial fibroelastosis on echocardiogram always associated with cardiac dysfunction 2) maternal antibodies to 52kD SSA/Ro, 60kD SSA/Ro, or 48kD SSB/La. In 96%, 2nd or 3rd degree block was present. Cardiac NL subjects were genotyped using the Illumina 370K SNP platform and merged with 3351 Illumina out-of-study controls from SLEGEN (Harley 2008). Standard quality control and admixture adjusted tests of association were computed. Results: The HLA region (6p21) showed strong evidence of association. Two HLA SNPs in high linkage disequilibrium were rs3135353 (p_dom= 3.99E-08; OR = 2.87; 2.8 kb from HLA-DRB5) and rs3129963 (p_dom= 8.96E-07; OR = 2.57; 75 kb from HLA-DRA). Outside the HLA region, rs1810636 near 20p13 showed significant association to cardiac NL (p_rec= 1.53E-12, OR = 4.07). In proximity to this region are the DNA sequences of five noncoding RNAs that associate with the spliceosome. An additional suggestive association at 5q11.2 includes rs 2432143 (p_dom=5.87E-05, OR=2.28) within the integrin, alpha 1, a receptor involved in cell-cell adhesion, which may play a role in inflammation and fibrosis. Importantly, none of the fifteen prominent non-HLA polymorphisms reported in SLE association studies were associated with cardiac NL at less than 1E-5. Conclusion: These analyses are the first genome-wide association study for cardiac NL and identify several strong statistical associations. These associations, including the HLA region, corroborate the genetic influences on cardiac NL and may provide a basis for exuberant fibrosis of the conduction system
— id: 130347, year: 2009, vol: 60, page: 1230, stat: Journal Article,

Preventive IVIG therapy for congenital heart block (PITCH)
Friedman D.M.; Llanos C.; Izmirly P.M.; Kim M.Y.; Buyon J.P.
2009 ;60:1251-1251, Arthritis & rheumatism
Background: The recurrence rate of anti-SSA/Ro associated congenital heart block (CHB) is 17%. Reversal of 3^rd degree block has never been achieved. Prophylactic IVIG was considered based on two presumed mechanisms of efficacy, a) saturation of FcRn to accelerate maternal IgG catabolism and decrease placental transport b) elevation of macrophage FcRIIB expression to attenuate inflammatory fetal responses. Purpose: To evaluate IVIG efficacy and safety as a preventive therapy for CHB. Methods: A multicenter open-label study based on Simon's 2-stage optimal design was initiated. Enrollment criteria included: maternal anti-SSA/Ro antibody, a previous child with CHB/rash, nd or 3^rd degree CHB in three fetuses. Results: Twenty mothers were enrolled. Sixteen children had normal PR intervals throughout the study and no manifestations of neonatal lupus. One child developed a transient rash consistent with neonatal lupus and had normal PR intervals during pregnancy and normal EKG at birth. However, the pre-determined stopping rule was reached. CHB was detected in three fetuses, at 19, 20 and 25 weeks; none followed an abnormal PR interval. One of these mothers had two previous children with CHB. Antibody titers assessed before every IVIG infusion, and at 28 wks, 34 wks and delivery were compared with values obtained at baseline. No significant changes in maternal antibody titers to SSA/Ro, SSB/La, or Ro52 were detected over the course of therapy or at delivery (P>0.05 for all comparisons). There were no changes in maternal blood pressure, severe headaches, rashes, fever or any other adverse effects related to the infusions. Neonatal weight, height, and head circumference were derived from gestational age -specific growth curves to correct for prematurity when necessary. Four (21%) of the newborns, two with CHB and two healthy, were small for gestational age (<10th centile) and 3 healthy babies (16%) were born prematurely (<37 weeks of gestation). Conclusion: IVIG at doses consistent with replacement does not prevent the recurrence of CHB or reduce maternal antibody titers. Having established safety with this protocol and feasibility of patient enrollment, subsequent studies should address the efficacy of IVIG at higher doses to exploit an anti-inflammatory effect
— id: 130338, year: 2009, vol: 60, page: 1251, stat: Journal Article,

Prospective evaluation of fetuses with autoimmune-associated congenital heart block followed in the PR Interval and Dexamethasone Evaluation (PRIDE) Study
Friedman, Deborah M; Kim, Mimi Y; Copel, Joshua A; Llanos, Carolina; Davis, Claudine; Buyon, Jill P
2009 Apr 15;103(8):1102-1106, American journal of cardiology
We evaluated the efficacy of dexamethasone (DEX) in anti-SSA/Ro-exposed fetuses newly diagnosed with congenital heart block. Previous use of DEX has been anecdotal with varying reports of therapeutic benefit. This was a multicenter, open-label, nonrandomized study involving 30 pregnancies treated with DEX (22 with third-degree block, 6 with second-degree block, 2 with first-degree block) and 10 untreated (9 with third-degree block, 1 with first-degree block). Initial median ventricular rates, age at diagnosis, and degree of cardiac dysfunction were similar between groups. Six deaths occurred in the DEX group. There was no reversal of third-degree block with therapy or spontaneously. In fetuses treated with DEX, 1/6 with second-degree block progressed to third-degree block and 3 remained in second-degree block (postnatally 1 paced, 2 progressed to third degree); 2 reverted to normal sinus rhythm (NSR; postnatally 1 progressed to second degree). DEX reversed the 2 fetuses with first-degree block to NSR by 7 days with no regression at discontinuation. Absent DEX, the 1 with first-degree block detected at 38 weeks had NSR at birth (overall stability or improvement in 4 of 8 in the DEX group vs 1 of 1 in the non-DEX group). Median gestational birth age was 37 weeks in the DEX group versus 38 weeks in the non-DEX group (p = 0.019). Prematurity and small size for gestational age were restricted to the DEX group. Pacemaker use and growth parameters at birth and 1 year were similar between groups. In conclusion, these data confirm the irreversibility of third-degree block and progression of second- to third-degree block despite DEX. A potential benefit of DEX in reversing first- or second-degree block was supported in rare cases but should be weighed against potential steroid side effects such as growth restriction
— id: 114631, year: 2009, vol: 103, page: 1102, stat: Journal Article,

Anatomic and Pathologic Findings in Hearts From Fetuses and Infants With Cardiac Manifestations of Neonatal Lupus
Friedman, DM; Llanos, C; Izmirly, PM; Clancy, RM; Buyon, JP
2009 NOV 3 ;120(18):S578-S578, Circulation
— id: 106972, year: 2009, vol: 120, page: S578, stat: Journal Article,

Preventive IVIG Therapy for Congenital Heart Block (PITCH) Study
Friedman, DM; Llanos, C; Izmirly, PM; Phoon, CK; Kim, MY; Buyon, JP
2009 NOV 3 ;120(18):S577-S578, Circulation
— id: 106971, year: 2009, vol: 120, page: S577, stat: Journal Article,

Association of soluble E-selectin and adiponectin with carotid plaque, independent of clinical activity, in patients with systemic lupus erythematosus
Izmirly P.M.; Reynolds H.R.; Rivera T.L.; Kim M.Y.; Tunick P.A.; Buyon J.P.; Clancy R.M.
2009 ;60:1562-1562, Arthritis & rheumatism
Purpose: The mechanisms underlying premature atherosclerosis in SLE are not understood. The endothelium merits focus since it provides the physiologic boundary which limits extravasation and diapedesis of inflammatory cells. Methods: One hundred and nineteen patients with SLE, predominantly non-Caucasian, and 71 healthy controls matched for age, sex and race, underwent carotid ultrasonography and donated blood for evaluation of circulating endothelial cells (CEC), soluble endothelial protein C receptor (sEPCR) and gene polymorphism at A6936G, soluble E-selectin, and adiponectin. Results: Carotid plaque was more prevalent among patients than controls (43% vs 17%, p=0.0002). Mean CCA IMT was greater in patients compared to controls (0.59mm+/-0.19 vs 0.54mm+/-0.11, p=0.03). Levels of CEC (19 vs 3 CECs/mL, p<0.0001) and sE-selectin (64 vs 36 ng/ml, p<0.0001) were significantly elevated in patients compared to controls. Unexpectedly, adiponectin was also significantly higher in patients compared to controls (16 ug/mL versus 11 ug/mL, p=0.0001) but no differences were seen in the levels of sEPCR or the distribution of genotype. Independent predictors of plaque status using logistic regression models included: age (p<0.0001; OR=2.1 per 10 year increase; 95% CI: 1.5-3.0), SLE status (p=0.015; OR=3.4 for SLE vs control; 95% CI: 1.3-9.1), sE-selectin (p=0.016; OR=1.2 per 10 unit increase; 95% CI: 1.0-1.4) and adiponectin (p=0.050; OR=1.5 per 10 unit increase; 95% CI: 1.0-2.4). Comparing SLE patients with and without plaque, there were no differences in cardiac CRP, complement, anti-dsDNA ab, CEC, sEPCR levels and EPCR SNP. However, sE-selectin and adiponectin levels were significantly higher in SLE with plaque compared to those without (sE-selectin 78 vs 52 ng/ml; p=0.006; adiponectin 18 vs 14 ug/ml; p=0.033). The estimated odds ratios for plaque in the final logistic regression model were: OR_selectin= 1.3 per 10 ng/ml increase (95% CI: 1.1-1.5) and OR_adiponectin=1.8 per 10 ug/ml increase (95% CI: 1.1-3.0). SELENA-SLEDAI scores were similar between groups, and the proportion of patients with SLEDAI<= 4 did not segregate with the absence of plaque. Neither past nor current medications significantly associated with plaque. In the stable subjects (SLEDAI <=4), age (p=0.007), sE-selectin (p=0.02) and adiponectin (p=0.02) remained associated with plaque. The prevalence of plaque was greatest in the stable patients with high sE-selectin plus high adiponectin (55%; p =0.0009) confirming the multivariable analyses. Sixty-two patients donated blood at a second visit. High sE-selectin and adiponectin were sustained in plaque patients compared to non-plaque patients (p=0.0009 and p=0.0011 respectively). Conclusion: These results confirm that SLE patients, irrespective of race, are at increased risk for premature atherosclerosis and support the hypothesis that endothelial perturbation is contributory even in the absence of clinically measurable disease activity
— id: 130319, year: 2009, vol: 60, page: 1562, stat: Journal Article,

Expression of endothelial protein C receptor in cortical peritubular capillaries associates with a poor clinical response in lupus nephritis
Izmirly, Peter M; Barisoni, Laura; Buyon, Jill P; Kim, Mimi Y; Rivera, Tania L; Schwartzman, Julie S; Weisstuch, Joseph M; Liu, David T; Bernstein, Stephen; Tseng, Chung-E; Belmont, Howard M; Esmon, Charles T; Merrill, Joan T; Askanase, Anca D; Thomas, David B; Clancy, Robert M
2009 May;48(5):513-519, Rheumatology (Oxford)
OBJECTIVE: To study the membrane expression of endothelial protein C receptor (mEPCR) in the renal microvasculature in lupus nephritis (LN) as a potential marker of injury and/or prognostic indicator for response to therapy. METHODS: mEPCR expression was analysed by immunohistochemistry in normal kidney and in 59 biopsies from 49 patients with LN. Clinical parameters were assessed at baseline, 6 months and 1 year. RESULTS: mEPCR was expressed in the medulla, arterial endothelium and cortical peritubular capillaries (PTCs) in all biopsies with LN but not in the cortical PTCs of normal kidney. Positive mEPCR staining in >25% of the PTCs was observed in 16/59 biopsies and associated with poor response to therapy. Eleven (84.6%) of 13 patients with positive staining for mEPCR in >25% of the PTCs and follow-up at 6 months did not respond to therapy, compared with 8/28 (28.6%) with mEPCR staining in < or =25% PTCs, P = 0.0018. At 1 year, 10 (83.3%) of 12 patients with positive mEPCR staining in >25% of the PTCs did not respond to therapy (with two progressing to end-stage renal disease) compared with 8/24 (33.3%) with positive staining in < or =25% of the PTCs, P = 0.0116. Although tubulo-interstitial damage (TID) was always accompanied by mEPCR, this endothelial marker was extensively expressed in the absence of TID suggesting that poor response could not be attributed solely to increased TID. mEPCR expression was independent of International Society of Nephrology/Renal Pathology Society class, activity and chronicity indices. CONCLUSION: Increased mEPCR expression in PTCs may represent a novel marker of poor response to therapy for LN
— id: 100419, year: 2009, vol: 48, page: 513, stat: Journal Article,

Modulation of TLR7 and TLR8 activation in human macrophages
Komissarova E.V.; Kandimalla E.R.; Agrawal S.; Clancy R.M.; Buyon J.P.
2009 ;60:141-141, Arthritis & rheumatism
Purpose: Two percent of neonates born to mothers with SSA/Ro autoantibodies have congenital heart block (CHB). It has been hypothesized that macrophage engagement of Toll-like receptors (TLR) via binding to the ssRNA moiety of the target autoantigen exposed on the surface of the fetal cardiocytes is involved in the pathogenesis. Using a RNA-based TLR agonist and an antagonist of TLR7 and TLR9, activation of TLR was studied by TNFalpha release. Method: Macrophages were derived from CD14+ monocytes of healthy donors cultured in suspension with growth medium RPMI1640/10% FBS and 10ng/ml GM-CSF for at least 7 days. Cells were plated at 4x10⁵ per well into 12-well plates and adhered macrophages were treated with TLR7 or TLR8 agonists in the absence or presence of antagonist for 24 hours. The levels of TNFalpha in culture supernatants were measured by ELISA. Cytotoxicity of compounds for macrophages was assessed in MTS assay. Expression of TLR7 and TLR8 mRNA was determined in RT-PCR. Results: Macrophages from different donors showed variability in the levels of TLR7 and TLR8 mRNA expression and corresponding variability in response to TLR agonists.TLR7 agonist R837 (imiquimod, Invivo Gen) at 5mg/ml induced statistically significant TNFalpha release from 31.6 to 457.7+/-85.9pg/ml (p=0.0033, N=7). RNA-based TLR8 agonist at 20mg/ml induced statistically significant TNFa release from 31.6 to 621+/- 124.6pg/ml (p=0.0029, N=7). The novel TLR antagonist at 80mg/ml (nontoxic for macrophages) reduced the induced TNFalpha release by TLR8 agonist (20mug/ml) from 621+/-124.6 to 11.4+/-5.9pg/ml (N=3). Endosomal acidification neutralizing agent, chloroquine at a nontoxic concentration of 5mg/ml inhibited the induced TNFa secretion by R837 (5mg/ml) from 457.7+/-85.9 to 184.6+/-31.07 (p=0.0086, N=6) and by TLR8 agonist (20mug/ml) from 621+/-124.6 to 4.4 pg/ml (N=2). No toxicity was seen in macrophages exposed for 24 hours to TLR antagonist at concentrations up to 100 mug/ml (MTS assay). In contrast, after a 24-hour exposure to 21mg/ml chloroquine only 75% of the macrophages were viable in MTS assay. The LC₅₀ value for chloroquine was 40mug/ml. Conclusion: Variability of TLR7 and TLR8 expression in isolated human macrophages from different donors may be relevant to the inflammatory cascade to CHB and discordant disease in fetuses exposed to anti-Ro antibodies. Given the profound TLR inhibition, antimalarial compounds, already in use during human pregnancy, and TLR antagonist studied herein may be relevant candidates for study in the prevention of disease
— id: 130333, year: 2009, vol: 60, page: 141, stat: Journal Article,

Methodologic issues in the validation of putative biomarkers and surrogate endpoints in treatment evaluation for systemic lupus erythematosus
Liang, Matthew H; Simard, Julia F; Costenbader, Karen; Dore, Benjamin T; Ward, Michael; Fortin, Paul R; Illei, Gabor G; Manzi, Susan; Mittleman, Barbara; Buyon, Jill; Gupta, Samardeep; Abrahamowicz, Michal
2009 Mar;9(1):108-112, Endocrine, metabolic & immune disorders drug targets
No new drugs have been approved for the treatment of systemic lupus erythematosus (SLE) by the Food and Drug Administration for the last 30 years. One barrier has been the lack of validated biomarkers and surrogate endpoints. Validation of SLE biomarkers in the past have been methodologically flawed. We put forth a conceptual framework and five critical criterion for validating putative biomarkers and bio-surrogates in this heterogeneous multi-system disease with protean manifestations. Using the example of a putative biomarker for end-stage lupus nephritis, we performed computer simulations for planning a biomarker bio-repository to support the validation process. 'Random time window' sampling where a biomarker is obtained in an interval randomly selected from the total follow-up time for that subject creates survival bias. This can be avoided by the 'fixed calendar window' design, in which biomarkers are measured within the same, pre-specified period for all cohort members who remain at risk during that period. In lupus nephritis where the incidence rate of end-stage renal disease is relatively low, to accumulate 300 instances of end-stage renal disease, at risk patients would have to be followed for about 5,000 person-years, implying 500 subjects followed, on average, for about 10 years. Increasing the number of biomarker determinations per subject from one to five reduces the required number of subjects by 10-15%, while further increases in the number of observations per subject yielded much smaller gains. The large numbers of subjects required for a bio-repository, makes it essential to maximize the efficiency of study designs and analyses and provides the strongest rationale for collaboration and the use of standardized measures to ensure comparability
— id: 120805, year: 2009, vol: 9, page: 108, stat: Journal Article,

Antibody Reactivity to {alpha}-Enolase in Mothers of Children with Congenital Heart Block
Llanos, Carolina; Chan, Edward K L; Li, Songqing; Abadal, Grant X; Izmirly, Peter; Byrne, Caroline; Clancy, Robert M; Buyon, Jill P
2009 Mar;36(3):565-569, Journal of rheumatology
OBJECTIVE: To evaluate the frequency of anti-alpha-enolase antibodies in the sera of mothers whose children have congenital heart block (CHB), given provocative results in which alpha-enolase, a membrane protein, was recognized by monoclonal antibodies reactive with the peptide p200 of 52 kDa Ro/SSA in a neonatal rat heart library. METHODS: An ELISA using a recombinant alpha-enolase protein was developed. Sera from 100 anti-Ro52+ CHB mothers in the Research Registry for Neonatal Lupus, 50 patients with systemic lupus erythematosus (SLE; 7 anti-Ro52+), and 48 healthy controls were tested for anti-alpha-enolase reactivity. RESULTS: There were no significant differences in the median values obtained from CHB mothers, patients with SLE, or controls at each of the dilutions tested. Only 7 (7%) at 1:100 dilution and 2 (2%) at 1:1000 dilution of 100 CHB sera were 3 standard deviations above the mean value obtained for controls. Preincubation with recombinant Ro52 did not inhibit anti-alpha-enolase reactivity. CONCLUSION: The low frequency of anti-alpha-enolase antibodies in the sera of CHB mothers and the absence of apparent cross-reactivity with Ro52 suggest that antibodies to Ro52 are not likely to mediate CHB via binding to alpha-enolase
— id: 94464, year: 2009, vol: 36, page: 565, stat: Journal Article,

Recurrence rates of cardiac manifestations associated with neonatal lupus and maternal/fetal risk factors
Llanos, Carolina; Izmirly, Peter M; Katholi, Margaret; Clancy, Robert M; Friedman, Deborah M; Kim, Mimi Y; Buyon, Jill P
2009 Oct;60(10):3091-3097, Arthritis & rheumatism
OBJECTIVE: Identifying the frequency of recurrent cardiac manifestations of neonatal lupus (NL) in a second child is critical to understanding the pathogenesis of anti-SSA/Ro-mediated injury and would improve counseling strategies regarding future pregnancies and power the design of clinical prevention trials. Accordingly, this study was undertaken to address the recurrence rates of cardiac NL and associated risk factors in a large US-based cohort. METHODS: Families enrolled in the Research Registry for Neonatal Lupus were evaluated for rates of recurrence of cardiac NL and potential risk factors, with a focus on pregnancies immediately following the birth of an affected child. RESULTS: The overall rate of recurrence of cardiac NL in 161 pregnancies of 129 mothers with anti-SSA/Ro antibodies was 17.4% (95% confidence interval 11.1-23.6%). Analysis of the potential risk factors among 129 mothers with a pregnancy immediately following the birth of a child with cardiac NL showed that the maternal diagnosis was not associated with the outcome in a subsequent pregnancy. In this group, 23% of mothers who were either asymptomatic or had an undifferentiated autoimmune syndrome, compared with 14% of mothers with systemic lupus erythematosus or Sjogren's syndrome, had a second child with cardiac NL (P = 0.25). The recurrence rate was not statistically significantly different in mothers who had taken steroids compared with those who had not taken steroids (16% versus 21%; P = 0.78). The antibody status of the mother was not predictive of outcome in subsequent pregnancies. Moreover, death of the first child with cardiac NL was not predictive of recurrence of cardiac NL in a subsequent pregnancy (P = 0.31). The risk of cardiac NL was similar between male and female children (17.2% versus 18.3%; P = 1.0). CONCLUSION: In this cohort, the overall recurrence rate for cardiac NL was 17%. The recurrence rate appeared to be unaffected by maternal health, use of steroids, antibody status, severity of cardiac disease in the first affected child, or sex of the subsequent child
— id: 104348, year: 2009, vol: 60, page: 3091, stat: Journal Article,

Flare assessment in systemic lupus erythematosus (SLE) patients treated with rituximab in the phase II/III EXPLORER trial
Merrill J.T.; Buyon J.P.; Furie R.; Latinis K.M.; Gordon C.; Hsieh H.; Brunetta P.
2009 ;60:925-925, Arthritis & rheumatism
Purpose: SLE patients (pts) with moderate to severe disease activity (>= 1 BILAG A or >= 2 BILAG B domain scores) despite background immunosuppressives and corticosteroids, were randomized to placebo (PLA) or rituximab (RTX). Although differences in primary and secondary outcome measures were not observed, an exploratory analysis was performed to evaluate the impact of RTX on the intensity and frequency of disease flares. Our objectives were to assess in those pts who achieved response whether RTX affected: 1) time to moderate or severe flares, 2) annualized flare rates, and 3) prednisone usage during flares. Methods: The BILAG index was scored every 4 wks for 52 wks after the first study drug treatment. Only pts who achieved response (BILAG C, D, or E scores for all domains before wk 52) were included in this analysis. A severe flare was defined as >= 1 BILAG A or > 3 BILAG B domain scores in a pt who previously achieved response; an A flare was defined as >= 1 new BILAG A domain scores; a moderate flare was defined as 2 BILAG B domain scores following response. Kaplan-Meier estimates were used to assess time to flare. Annualized flare rates were calculated using the number of flares a patient experienced during the study; the p-value was based on Poisson regression model. Only flares that occurred after the protocol-mandated prednisone taper at 12 wks were incorporated into the analysis of prednisone usage during flares. Results: A response was achieved in 58 (66.0%) of 88 PLA-treated and 127 (75.1%) of 169 RTX-treated pts during the study, and these patients qualified for subset analysis. Whereas the times to first moderate or severe flare were not different in the two treatment groups, there was a trend towards a prolonged time to first A flare in the RTX group (HR=0.612; p=0.052). Annualized severe and moderate flare rates were similar in both groups, but the mean annualized A flare rate in the RTX group was significantly lower than in the PLA group (0.86 vs 1.41; p=0.038). Prednisone was increased in 20 of 101 (20.7%) of the A flares and was not significantly different between the RTX and PLA groups (24% vs 14%; p=0.204). Pts with the highest initial prednisone dosing (> 60 mg/day) had lower treatment rates (14.0%). In one month, 44/81 (54.3%) of the untreated A flare improved to B or better. Conclusion: There was no difference between PLA and RTX in preventing moderate or severe flares after a response was achieved. However, RTX treatment was associated with a lower annualized A flare rate, and a trend towards prolonging the time to A flare. Prednisone rescue following A flares was low compared to observational cohorts, possibly reflecting a reluctance to reinitiate prednisone, or reduced BILAG A severity, in this trial
— id: 130329, year: 2009, vol: 60, page: 925, stat: Journal Article,

Connective tissue diseases: What does the death of Riquent hold for the future of SLE?
Merrill, Joan T; Buyon, Jill P
2009 Jun;5(6):306-307, Nature reviews. Rheumatology
In clinical trials testing new treatments for systemic lupus erythematosus (SLE), the failure of the 15-year drug development program for riquent (abetimus sodium) is the latest in a string of disappointments for a disease that has seen no new drugs approved in over 50 years
— id: 114632, year: 2009, vol: 5, page: 306, stat: Journal Article,

Revision of the SELENA flare index
Petri M.; Buyon J.; Kalunian K.C.; Urowitz M.B.; Strand V.; Merrill J.; Hahn B.H.
2009 ;60:902-902, Arthritis & rheumatism
Purpose: The original SELENA Flare Index (SFI) performed well in the SELENA trials to separate severe from mild/moderate flares. However, in current clinical trials it is important to also separate mild from moderate flares. Method: As part of the Lupus Foundation of America's International Flare Definition and Validation initiative, the SELENA Flare Index (SFI) was revised through a series of consensus conference calls. There was an initial review and discussion of the strengths and weaknesses of the SFI in SELENA and industry trials. Each call reviewed organ specific definitions of mild, moderate and severe flare. Definitions were discussed and re-reviewed until consensus was reached. Results:The revised SFI is organ-system based, and is not linked to the SLEDAI. For each organ system, suggested clinical manifestations are given for each category, but the intended treatment decision (if it is higher) overrides the clinical description in each category. Specifically, a mild flare is assigned if there is either no treatment, or initiation of hydroxychloroquine, prednisone <= 7.5 mg/day or a non-immunosuppressive therapy. Definition of a Moderate flare is consistent with use of prednisone > 7.5 mg/day but less than 0.5 mg/kg/day, or immunosuppressive therapy (other than cyclophosphamide), and Severe flare prednisone (or equivalent) >= 0.5 mg/kg/day, cyclophosphamide, biologic treatment, or hospitalization. Three mild flares are considered a moderate flare. Conclusion: The new SFI is organ system-based. The choice of recommended treatment overrides the clinical definition when the treatment choice is higher
— id: 130341, year: 2009, vol: 60, page: 902, stat: Journal Article,

Disease progression in mothers of children enrolled in the Research Registry for Neonatal Lupus
Rivera, T L; Izmirly, P M; Birnbaum, B K; Byrne, P; Brauth, J B; Katholi, M; Kim, M Y; Fischer, J; Clancy, R M; Buyon, J P
2009 Jun;68(6):828-835, Annals of rheumatic diseases
OBJECTIVE: /B> To evaluate autoimmune disease progression in asymptomatic and pauci-symptomatic mothers of children with neonatal lupus (NL). METHODS: Clinical information on mothers enrolled in the Research Registry (RRNL) was obtained from medical records. Genotyping was performed for -308A/G TNFalpha, 869T/C TGFbeta, and -889C/T IL1alpha. RESULTS: Of the 321-mothers enrolled, 229 had at least six months of follow-up. Twenty-six of the fifty-one mothers who were asymptomatic at the NL-child's birth progressed: 12 developed pauci-undifferentiated autoimmune syndrome (UAS), two poly-UAS, seven SS, four SLE and one SLE/SS. The median time to develop any symptom was 3.15 years. Sixteen of the 37 mothers classified as pauci-UAS at the NL-child's birth progressed: five developed poly-UAS, six SS, four SLE, and one SLE/SS. Of the pauci-UAS mothers enrolled within one year, the median time to progression was 6.7 years. Four mothers developed lupus nephritis (two asymptomatic, two pauci-UAS). The probability of an asymptomatic mother developing SLE by 10 years was 18.6%, and developing probable/definite SS was 27.9%. NL-manifestations did not predict disease progression in an asymptomatic mother. Mothers with anti-SSA/Ro and anti-SSB/La were nearly twice as likely to develop an autoimmune disease as mothers with anti-SSA/Ro only. Only TGFbetaT/T was significantly higher in SLE-mothers compared to asymptomatic-mothers (p=0.03). CONCLUSION: Continued follow-up of asymptomatic NL-mothers is warranted since nearly half progress, albeit few develop SLE. While the anti-SSB/La antibodies may be a risk factor for progression, further work is needed to determine reliable biomarkers in otherwise healthy women with anti-SSA/Ro antibodies identified solely because of an NL-child
— id: 94465, year: 2009, vol: 68, page: 828, stat: Journal Article,

Current therapies for lupus nephritis in an ethnically heterogeneous cohort
Rivera, Tania L; Belmont, H Michael; Malani, Seema; Latorre, Melissa; Benton, Lauri; Weisstuch, Joseph; Barisoni, Laura; Tseng, Chung-E; Izmirly, Peter M; Buyon, Jill P; Askanase, Anca D
2009 Feb;36(2):298-305, Journal of rheumatology
OBJECTIVE: To evaluate responses to mycophenolate mofetil (MMF) and intravenous cyclophosphamide (CYC) in lupus nephritis in a multiethnic population. METHODS: This was a retrospective study of all patients with systemic lupus erythematosus (SLE) that underwent kidney biopsy at New York University Medical Center. Patients with followup of at least 6 months were included. Clinical response was defined as complete (return to +/- 10% of normal) or partial (improvement of 50% in abnormal renal measurements). RESULTS: Ninty-nine patients were included in the study: 86% females, 86% non-Caucasian, age 34.2 +/- 1.1 years, 62% with proliferative nephritis (PN; ISN/RPS-III and IV), and 32% with membranous nephritis (MN; ISN/RPS-V). Of the 70 patients with PN, 37 were treated with CYC and 33 with MMF. The baseline characteristics of the 2 treatment groups were different in the incidence of ISN/RPS-IV, values of serum creatinine and serum albumin, and type of insurance (p < 0.05). The response rate was greater in the MMF than in the CYC group (70% vs 41%). Responses to MMF were different in Asians (11/11), Caucasians (4/5), African Americans (3/5), and Hispanics (5/11). Responses to CYC had a similar distribution (Asians 6/10, Caucasians 4/5, African Americans 4/9, Hispanics 1/11). In the MN group (N = 23) responses were similar to the PN group (73% MMF and 38% CYC). After adjusting for race, serum creatinine, serum albumin, type of insurance, and class of nephritis, in a logistic regression model, response to MMF was superior to CYC: OR 6.2 (95% CI 1.9-20.2). Hispanics had worse outcome than Caucasians (OR 0.17). Longterm followup suggested no difference in maintenance with MMF or CYC. CONCLUSION: After controlling for the fact that less severe nephritis is preferentially treated with MMF, we found overall that response to MMF was superior to CYC. In this US population, ethnicity was observed to have an influence on response
— id: 93732, year: 2009, vol: 36, page: 298, stat: Journal Article,

Longitudinal evaluation of idiotypic and anti-idiotypic response to the major epitope of La/SSB autoantigen in mothers enrolled in the PITCH (Preventive IVIG Therapy for Congenital Heart Block) study
Routsias Jg.; Kyriakidis N.; Llanos C.; Clancy R.M.; Buyon J.; Tzioufas A.G.
2009 ;60:1586-1586, Arthritis & rheumatism
Purpose: To investigate the effect of IVIg on the idiotype (id)-antiidiotype (anti-id) network in pregnant women with anti-Ro/SSA and anti-La/SSB antibodies who had a previous history of a child with CHB (high risk pregnancies). Method: The interaction of IVIg with the synthetic peptides was investigated by direct ELISA assays as well as inhibition experiments. Ten mothers who previously gave birth to a child with CHB were treated during a subsequent pregnancy with IVIG using a protocol in the Preventive IVIG Therapy for Congenital Heart Block (PITCH) Study. All individuals exhibited ELISA reactivity against epitope 349-364 of La/SSB in their initial samples. Sequential sera were drawn from all mothers during pregnancy (before each IVIg infusion at 12, 15, 18, 21, 25 and 28, 34, delivery), and evaluated for antibodies against the epitope 349-364 (pep) of La/SSB (idiotypic), as well as its complementary epitope (cpep) and anti-349-364 Fab2 fragments (both anti-idiotypic). Anti-349-364 reactivity was also evaluated after removal of anti-Id antibodies. Results: IVIg contains low affinity antibodies, most probably polyreactive antibodies, binding to both pep and cpep. The reactivity of anti-cpep (anti-id) antibodies appeared to be higher, compared to anti-pep antibodies. A partial inhibition of idiotype-antiidiotype binding (30%), was observed after IVIg treatment of purified antibodies. Following dosing of IVIg the maternal antibody titres to the major epitope of La/SSB decreased by 20% to 50% in four patients, remained stable in five and increased in one by 30%. In contrast, there was a substantial increase of anti-Id reactivity in all patients, ranging from 20% to 250%. After removal of anti-Id antibodies the Id reactivity was increased (up to 300%) in all patients. None of the children developed any conduction abnormality. Conclusion: This study supports the novel finding that IVIg administration decreases autoantibody titres and enhances the anti-idiotypic antibody response in pregnant women with anti-Ro/SSA and anti-La/SSB antibodies
— id: 130332, year: 2009, vol: 60, page: 1586, stat: Journal Article,

What's in a name?
Buyon, J P; Brucato, A; Friedman, D M
2008 May;67(5):732-732, Annals of rheumatic diseases
— id: 93300, year: 2008, vol: 67, page: 732, stat: Journal Article,

Dying right to live longer: positing apoptosis as a link between maternal autoantibodies and congenital heart block
Buyon, J P; Clancy, R M
2008 ;17(2):86-90, Lupus
The association of isolated congenital heart block (CHB) with maternal autoantibodies to SSA/Ro and SSB/La ribonucleoproteins is approaching the predictable, even in mothers who are completely asymptomatic. Indeed, this model of passively acquired autoimmunity offers an exceptional opportunity to examine the effector arm of immunity and define the pathogenicity of an autoantibody in mediating tissue injury. The study of CHB exemplifies not only translational research, which inherently draws upon clinical observations and explores them in the laboratory, but 'integrational' research which attempts to fit critical clinical and basic observations together, even those seemingly at odds. The spectrum of conduction abnormalities includes second and third-degree block, but injury can extend to the myocardium and endocardium, in rare cases without AV nodal dysfunction. The rarity of disease continues to drive the search for factors (fetal and environmental) that might amplify the effects of the maternal autoantibodies. The identification of exaggerated apoptosis, macrophage/myfibroblast crosstalk, TGF beta expression, and extensive fibrosis in the conducting system and in some cases surrounding myocardium in fetuses dying with CHB, provide in vivo support for several parallel lines of in vitro investigation. Specifically, the consideration of exaggerated apoptosis as the initial link between maternal antibody and tissue injury led to the observation that cardiocytes are capable of phagocytosing autologous apoptotic cardiocytes and anti-Ro/La antibodies inhibit this function. Recognizing that this perturbation of physiologic efferocytosis might divert uptake to professional Fc gamma R-bearing phagocytes fits well with experiments demonstrating macrophage secretion of pro-inflammatory and fibrosing cytokines when coincubated with apoptotic cardiocytes bound by Ro/La antibodies. While CHB is rare, its study should set precedent for defining the role of autoantibodies in driving end organ disease
— id: 78688, year: 2008, vol: 17, page: 86, stat: Journal Article,

What's in a name?
Buyon, JP; Brucato, A; Friedman, DM
2008 AUG ;67(8):229-257, Annals of rheumatic diseases
— id: 86850, year: 2008, vol: 67, page: 229, stat: Journal Article,

Utility of cardiac monitoring in fetuses at risk for congenital heart block: the PR Interval and Dexamethasone Evaluation (PRIDE) prospective study
Friedman, Deborah M; Kim, Mimi Y; Copel, Joshua A; Davis, Claudine; Phoon, Colin K L; Glickstein, Julie S; Buyon, Jill P
2008 Jan 29;117(4):485-493, Circulation
BACKGROUND: Anti-SSA/Ro-associated third-degree congenital heart block is irreversible, prompting a search for early markers and effective therapy. METHODS AND RESULTS: One hundred twenty-seven pregnant women with anti-SSA/Ro antibodies were enrolled; 95 completed an evaluable course in 98 pregnancies. The protocol included fetal echocardiograms performed weekly from 16 to 26 weeks' gestation and biweekly from 26 to 34 weeks. PR intervals >150 ms were considered prolonged, consistent with first-degree block. Ninety-two fetuses had normal PR intervals. Neonatal lupus developed in 10 cases; 4 were neonatal lupus rash only. Three fetuses had third-degree block; none had a preceding abnormal PR interval, although in 2 fetuses >1 week elapsed between echocardiographic evaluations. Tricuspid regurgitation preceded third-degree block in 1 fetus, and an atrial echodensity preceded block in a second. Two fetuses had PR intervals >150 ms. Both were detected at or before 22 weeks, and each reversed within 1 week with 4 mg dexamethasone. The ECG of 1 additional newborn revealed a prolonged PR interval persistent at 3 years despite normal intervals throughout gestation. No first-degree block developed after a normal ECG at birth. Heart block occurred in 3 of 16 pregnancies (19%) in mothers with a previous child with congenital heart block and in 3 of 74 pregnancies (4%) in mothers without a previous child with congenital heart block or rash (P=0.067). CONCLUSIONS: Prolongation of the PR interval was uncommon and did not precede more advanced block. There was a trend toward more congenital heart block in fetuses of women with previously affected offspring than those without previously affected offspring. Advanced block and cardiomyopathy can occur within 1 week of a normal echocardiogram without initial first-degree block. Echodensities and moderate/severe tricuspid regurgitation merit attention as early signs of injury
— id: 77788, year: 2008, vol: 117, page: 485, stat: Journal Article,

Decreased risk of anti-Ro/La associated congenital heart (CHB) in fetuses exposed to hydroxychloroquine (HCQ)
Izinirly, P; Kim, M; Le, P; Llanos, C; Katholi, M; Clancy, R; Salmon, J; Buyon, J
2008 SEP ;58(9):S810-S810, Arthritis & rheumatism
— id: 88572, year: 2008, vol: 58, page: S810, stat: Journal Article,

Widespread Upregulation of membrane endothelial protein c receptor in a subset of patients with active lupus nephritis
Izmirly, P; Yee, H; Rivera, T; Belmont, HM; Tseng, CE; Esmon, C; Braun, A; Askanase, A; Ginzler, EM; Buyon, J; Clancy, R
2008 SEP ;58(9):S950-S950, Arthritis & rheumatism
— id: 88579, year: 2008, vol: 58, page: S950, stat: Journal Article,

Pregnancy outcomes subsequent to a child with autoimmune associated congenital heart block (CHB) and associated fetal/maternal risk factors
Llanos, C; Izmirly, P; Katholi, M; Edberg, JC; Kimberly, RP; Clancy, R; Buyon, J
2008 SEP ;58(9):S873-S873, Arthritis & rheumatism
— id: 88575, year: 2008, vol: 58, page: S873, stat: Journal Article,

Serum type I interferon activity is dependent on maternal diagnosis in anti-SSA/Ro-positive mothers of children with neonatal lupus
Niewold, Timothy B; Rivera, Tania L; Buyon, Jill P; Crow, Mary K
2008 Feb;58(2):541-546, Arthritis & rheumatism
OBJECTIVE: The type I interferon (IFN) pathway is activated in many patients with systemic lupus erythematosus (SLE), and high serum levels of IFN are associated with anti-SSA/Ro autoantibodies. To investigate the clinical features associated with type I IFN production in vivo, we compared serum IFN activity in individuals with anti-SSA/Ro antibodies who were asymptomatic with that in individuals with clinical manifestations of SLE or Sjogren's syndrome (SS). METHODS: Antibody-positive sera from 84 mothers of children with manifestations of neonatal lupus were studied for type I IFN activity, using a functional reporter cell assay. Maternal health status was characterized as asymptomatic, SS, SLE, pauci-SLE, or pauci-SS, based on a screening questionnaire, telephone interview, and review of medical records. The prefix 'pauci-' indicates symptoms insufficient for a formal classification of the disease. RESULTS: Only 4% of asymptomatic mothers had high serum type I IFN activity, compared with 73% with pauci-SLE (P = 5.7 x 10(-5)), 35% with SLE (P = 0.011), and 32% of patients with SS (P = 0.032). One of the 4 patients with pauci-SS had high levels of IFN. The majority of patients for whom longitudinal data were available had stable type I IFN activity over time, and changes in IFN activity were not clearly accompanied by changes in the clinical diagnosis. CONCLUSION: Patients with SLE, patients with pauci-SLE, and patients with SS are more likely to have high serum IFN activity than asymptomatic individuals with SSA/Ro autoantibodies, suggesting that these autoantibodies are insufficient for activation of the type I IFN pathway, and that disease-specific factors are important for type I IFN generation in vivo
— id: 77787, year: 2008, vol: 58, page: 541, stat: Journal Article,

Oral contraceptives in systemic lupus erythematosus: the case for (and against)
Petri, M; Buyon, J P
2008 Aug;17(8):708-710, Lupus
— id: 114629, year: 2008, vol: 17, page: 708, stat: Journal Article,

Systemic lupus international collaborating clinics renal activity/response exercise: development of a renal activity score and renal response index
Petri, Michelle; Kasitanon, Nuntana; Lee, Shin-Seok; Link, Kimberly; Magder, Laurence; Bae, Sang-Cheol; Hanly, John G; Isenberg, David A; Nived, Ola; Sturfelt, Gunnar; van Vollenhoven, Ronald; Wallace, Daniel J; Alarcon, Graciela S; Adu, Dwomoa; Avila-Casado, Carmen; Bernatsky, Sasha R; Bruce, Ian N; Clarke, Ann E; Contreras, Gabriel; Fine, Derek M; Gladman, Dafna D; Gordon, Caroline; Kalunian, Kenneth C; Madaio, Michael P; Rovin, Brad H; Sanchez-Guerrero, Jorge; Steinsson, Kristjan; Aranow, Cynthia; Balow, James E; Buyon, Jill P; Ginzler, Ellen M; Khamashta, Munther A; Urowitz, Murray B; Dooley, Mary Anne; Merrill, Joan T; Ramsey-Goldman, Rosalind; Font, Josef; Tumlin, James; Stoll, Thomas; Zoma, Asad
2008 Jun;58(6):1784-1788, Arthritis & rheumatism
OBJECTIVE: To develop a measure of renal activity in systemic lupus erythematosus and use it to develop a renal response index. METHODS: Abstracted data from the medical records of 215 patients with lupus nephritis were sent to 8 nephrologists and 29 rheumatologists for rating. Seven nephrologists and 22 rheumatologists completed the ratings. Each physician rated each patient visit with respect to renal disease activity (none, mild, moderate, or severe). Using the most commonly selected rating for each patient as the gold standard, stepwise regression modeling was performed to identify the variables most related to renal disease activity, and these variables were then used to create an activity score. This activity score could then be applied to 2 consecutive visits to define a renal response index. RESULTS: The renal activity score was computed as follows: proteinuria 0.5-1 gm/day (3 points), proteinuria >1-3 gm/day (5 points), proteinuria >3 gm/day (11 points), urine red blood cell count >10/high-power field (3 points), and urine white blood cell count >10/high-power field (1 point). The chance-adjusted agreement between the renal response index derived from the activity score applied to the paired visits and the plurality physician response rating was 0.69 (95% confidence interval 0.59-0.79). CONCLUSION: Ratings derived from this index for rating of renal response showed reasonable agreement with physician ratings in a pilot study. The index will require further refinement, testing, and validation. A data-driven approach to create renal activity and renal response indices will be useful in both clinical care and research settings
— id: 95296, year: 2008, vol: 58, page: 1784, stat: Journal Article,

Systemic lupus international collaborating clinics renal activity/response exercise: comparison of agreement in rating renal response
Petri, Michelle; Kasitanon, Nuntana; Singh, Sukminder; Link, Kimberly; Magder, Laurence; Bae, Sang-Cheol; Hanly, John G; Nived, Ola; Sturfelt, Gunnar; van Vollenhoven, Ronald; Wallace, Daniel J; Alarcon, Graciela S; Adu, Dwomoa; Avila-Casado, Carmen; Bernatsky, Sasha R; Bruce, Ian N; Clarke, Ann E; Contreras, Gabriel; Fine, Derek M; Gladman, Dafna D; Gordon, Caroline; Kalunian, Kenneth C; Madaio, Michael P; Rovin, Brad H; Sanchez-Guerrero, Jorge; Steinsson, Kristjan; Aranow, Cynthia; Balow, James E; Buyon, Jill P; Ginzler, Ellen M; Khamashta, Munther A; Urowitz, Murray B; Dooley, Mary Anne; Merrill, Joan T; Ramsey-Goldman, Rosalind; Font, Josef; Tumlin, James; Stoll, Thomas; Zoma, Asad
2008 Jun;58(6):1789-1795, Arthritis & rheumatism
OBJECTIVE: To assess the degree to which physicians agree with each other and with ratings obtained with 3 existing responder indices, in rating the response to treatment of lupus nephritis. METHODS: Lupus nephritis patient medical records from 125 pairs of visits (6 months apart) were used to create renal response scenarios. Seven nephrologists and 22 rheumatologists rated each scenario as demonstrating complete response, partial response, same, or worsening. The plurality (most frequent) rating of renal response by the physicians was compared with the calculated score from the renal component of the British Isles Lupus Assessment Group (BILAG) index (original and updated [2004] version) and of the Responder Index for Lupus Erythematosus (RIFLE). The degree of agreement among the physicians was assessed by calculating intraclass correlation coefficients (ICCs). The degree of agreement between the plurality physician rating and ratings obtained with the established response indices was assessed using the kappa statistic. RESULTS: The ICC among all physicians was 0.64 (0.62 for nephrologists and 0.67 for rheumatologists). The chance-adjusted measure of agreement (kappa coefficient) between the plurality physician rating and the calculated score obtained using established indexes was 0.50 (95% confidence interval [95% CI] 0.38-0.61) for the RIFLE, 0.14 (95% CI 0.03-0.25) for the original BILAG, and 0.23 (95% CI 0.21-0.44) for the BILAG 2004. CONCLUSION: These findings indicate that rheumatologists as a group and nephrologists as a group have equal agreement in their rating of renal response. There was moderate agreement between plurality physician ratings and ratings obtained using the renal component of the RIFLE. Ratings of response using an index based on the original BILAG did not have good agreement with the plurality physician rating
— id: 95297, year: 2008, vol: 58, page: 1789, stat: Journal Article,

Longitudinal evaluation of anti-La idiotypic antibodies in mothers enrolled in the PITCH study
Routsias, J; Stea, E; Geffrard, L; Friedman, D; Llanos, C; Clancy, RC; Buyon, J; Tzioufas, AG
2008 SEP ;58(9):S804-S804, Arthritis & rheumatism
— id: 88571, year: 2008, vol: 58, page: S804, stat: Journal Article,

Antibodies to amino acid 200-239 (p200) of Ro52 as serological markers for the risk of developing congenital heart block
Strandberg, L; Winqvist, O; Sonesson, S-E; Mohseni, S; Salomonsson, S; Bremme, K; Buyon, J P; Julkunen, H; Wahren-Herlenius, M
2008 Oct;154(1):30-37, Clinical & experimental immunology
Maternal autoantibodies to the p200-epitope of Ro52 have been suggested to correlate with development of congenital heart block. The aim of the present study was to evaluate the clinical relevance and predictive value of p200-antibodies in high-risk pregnancies. Sera from 515 Finnish, Swedish and American women were included in the study. Sera originated from 202 mothers with an infant affected by second- or third-degree atrioventricular block (AVB), 177 mothers with rheumatic disease having infants with normal heart rate and female blood donors (n = 136). A novel serological assay for Ro52 p200-antibodies with intra- and inter-assay variability of 3% and 3.8% respectively was developed. Mothers of children affected by AVB II-III had significantly higher p200-antibody levels than mothers with rheumatic disease having children with normal heart rate (P < 0.001). In the Swedish cohort, a distinction between foetuses with normal conduction, AVB I, AVB II and III was possible. A significant difference in anti-p200 levels between AVB I and AVB II-III groups compared with foetuses with normal conduction (P < 0.05 and P < 0.01) was observed. Using p200-antibodies as a second step analysis in Ro52-positive pregnancies increased the positive predictive value for foetal cardiac involvement (AVB I, II or III) from 0.39 (0.27-0.51) to 0.53 (0.37-0.68). In conclusion, Ro52 p200-antibodies may occur in women with unaffected children, but levels are significantly higher in mothers of children with congenital heart block and are suggested as a relevant marker in evaluating the risk for foetal AV block
— id: 114630, year: 2008, vol: 154, page: 30, stat: Journal Article,

The autoimmune connection : essential information for women on diagnosis, treatment, and getting on with your life
Baron-Faust, Rita; Buyon, Jill P
Princeton NJ : Recording for the Blind & Dyslexic, 2007,
— id: 1681, year: 2007, vol: , page: , stat: ,

Neonatal lupus
Buyon, Jill P; Clancy, Robert M
Dubois' lupus erythematosus Philadelphia : Lippincott Williams & Wilkins, 2007,
— id: 4872, year: 2007, vol: , page: ?, stat: Chapter,

Role of hypoxia and cAMP in the transdifferentiation of human fetal cardiac fibroblasts: implications for progression to scarring in autoimmune-associated congenital heart block
Clancy, Robert M; Zheng, Ping; O'Mahony, Marguerita; Izmirly, Peter; Zavadil, Jiri; Gardner, Lawrence; Buyon, Jill P
2007 Dec;56(12):4120-4131, Arthritis & rheumatism
OBJECTIVE: Identification of isolated congenital heart block (CHB) predicts, with near certainty, the presence of maternal anti-SSA/Ro antibodies; however, the 2% incidence of CHB in first offspring of anti-SSA/Ro+ mothers, 20% recurrence in subsequent pregnancies, and discordance in identical twins suggest that an environmental factor amplifies the effect of the antibody. Accordingly, this study was carried out to explore the hypothesis that hypoxia potentiates a profibrosing phenotype of the fetal cardiac fibroblast. METHODS: Evidence of an effect of hypoxia was sought by immunohistologic evaluation of CHB-affected fetal heart tissue and by determination of erythropoietin levels in cord blood. The in vitro effect of hypoxia on gene expression and phenotype in fibroblasts derived from fetal hearts and lungs was investigated by Affymetrix arrays, quantitative polymerase chain reaction (PCR), immunofluorescence, and immunoblotting. RESULTS: In vivo hypoxic exposure was supported by the prominent intracellular fibroblast expression of hypoxia-inducible factor 1alpha in conduction tissue from 2 fetuses in whom CHB led to death. The possibility that hypoxia was sustained was suggested by significantly elevated erythropoietin levels in cord blood from CHB-affected, as compared with unaffected, anti-SSA/Ro-exposed neonates. In vitro exposure of cardiac fibroblasts to hypoxia resulted in transdifferentiation to myofibroblasts (a scarring phenotype), as demonstrated on immunoblots and immunofluorescence by increased expression of smooth muscle actin (SMA), an effect not seen in lung fibroblasts. Hypoxia-exposed cardiac fibroblasts expressed adrenomedullin at 4-fold increased levels, as determined by Affymetrix array, quantitative PCR, and immunofluorescence, thus focusing attention on cAMP as a modulator of fibrosis. MDL12,330A, an adenylate cyclase inhibitor that lowers the levels of cAMP, increased expression of fibrosis-related proteins (mammalian target of rapamycin, SMA, plasminogen activator inhibitor type 1, and type I collagen), while the cAMP activator forskolin attenuated transforming growth factor beta-elicited fibrosing end points in the cardiac fibroblasts. CONCLUSION: These findings provide evidence that hypoxia may amplify the injurious effects of anti-SSA/Ro antibodies. Modulation of cAMP may be a key component in the scarring phenotype. Further assessment of the susceptibility of cardiac fibroblasts to cAMP modulation offers a new research direction in CHB
— id: 75771, year: 2007, vol: 56, page: 4120, stat: Journal Article,

Epidemiology, etiology, detection, and treatment of autoantibody-associated congenital heart block in neonatal lupus
Friedman, Deborah M; Rupel, Ann; Buyon, Jill P
2007 May;9(2):101-108, Current rheumatology reports
Neonatal lupus syndrome is a model of passively acquired autoimmunity in which the pregnant woman's serum contains specific antibodies to 52 or 60 kd SSA/Ro and/or 48 kd SSB/La, which cross the placenta and are associated with the development of congenital heart block in the fetus and/or a transient rash or various liver and blood cell abnormalities in the newborn. To date, congenital heart block is a permanent condition that entails significant morbidity and mortality, with nearly all affected infants requiring pacemakers and with an 80% cumulative probability of survival at 3 years of age. An intensive search is on for the specific etiopathophysiology and for new clinical tools to approach and treat this disease
— id: 73111, year: 2007, vol: 9, page: 101, stat: Journal Article,

Neonatal lupus syndromes
Izmirly, Peter M; Rivera, Tania L; Buyon, Jill P
2007 May;33(2):267-85, vi, Rheumatic diseases clinics of North America
Neonatal lupus has become an important model of passively acquired autoimmunity since the seminal observation in the late 1970s that sera from nearly all mothers of children with isolated congenital heart block (CHB) contain specific autoantibodies
— id: 73110, year: 2007, vol: 33, page: 267, stat: Journal Article,

Estrogens and lupus: bubbling cauldron or another overrated Witches' Brew?
Lockshin, Michael D; Buyon, Jill P
2007 Apr;56(4):1048-1050, Arthritis & rheumatism
— id: 73521, year: 2007, vol: 56, page: 1048, stat: Journal Article,

Hormones and gender-related issues
Merrill, Joan T; Buyon, Jill P
Dubois' lupus erythematosus Philadelphia : Lippincott Williams & Wilkins, 2007,
— id: 4873, year: 2007, vol: , page: ?, stat: Chapter,

Different temporal expression of immunodominant Ro60/60 kDa-SSA and La/SSB apotopes
Reed, J H; Neufing, P J; Jackson, M W; Clancy, R M; Macardle, P J; Buyon, J P; Gordon, T P
2007 Apr;148(1):153-160, Clinical & experimental immunology
Opsonization of apoptotic cardiocytes by maternal anti-Ro/SSA and anti-La/SSB antibodies contributes to tissue injury in the neonatal lupus syndrome. The objective of the current study was to quantify the surface membrane expression of Ro/La components during different phases of apoptosis and map the Ro/La apotopes (epitopes expressed on apoptotic cells) bound by cognate antibodies. Multi-parameter flow cytometry was used to define early and late apoptotic populations and their respective binding by monospecific anti-Ro and anti-La IgGs. Anti-Ro60 bound specifically to early apoptotic Jurkat cells and remained accessible on the cell surface throughout early and late apoptosis. In contrast, anti-La bound exclusively to late apoptotic cells in experiments controlled for non-specific membrane leakage of IgG. Ro52 was not accessible for antibody binding on either apoptotic population. The immunodominant NH2-terminal and RNA recognition motif (RRM) epitopes of La were expressed as apotopes on late apoptotic cells, confirming recent in vivo findings. An immunodominant internal epitope of Ro60 that contains the RRM, and is recognized by a majority of sera from mothers of children with congenital heart block (CHB) and patients with primary Sjogren's syndrome, was also accessible as an apotope on early apoptotic cells. The distinct temporal expression of the immunodominant Ro60 and La apotopes indicates that these intracellular autoantigens translocate independently to the cell surface, and supports a model in which maternal antibody populations against both Ro60 and La apotopes act in an additive fashion to increase the risk of tissue damage in CHB
— id: 73543, year: 2007, vol: 148, page: 153, stat: Journal Article,

Elevated antiangiogenic factors predict preeclampsia in pregnant patients with SLE and/or APL antibodies
Salmon, JE; Guerra, M; Kim, M; Rana, S; Karumanchi, SA; Lockshin, M; Branch, W; Laskin, CA; Porter, TF; Petri, M; Merrill, J; Buyon, JP
2007 DEC ;56(12):4236-4236, Arthritis & rheumatism
— id: 87209, year: 2007, vol: 56, page: 4236, stat: Journal Article,

Corticosteroids in preventing severe lupus Hares: do all patients have the same risk? Comment on the article by Tseng et al - Reply
Tseng, CE; Buyon, JP; Kim, M; Belmont, HM; Abramson, SB
2007 JUN ;56(6):2099-2099, Arthritis & rheumatism
— id: 73344, year: 2007, vol: 56, page: 2099, stat: Journal Article,

Frequency of attention disorders in children exposed to anti-SSA/Ro-SSB/La antibodies
Askanase, AD; Katholi, MC; Harris, RR; Buyon, JP
2006 SEP ;54(9):S163-S163, Arthritis & rheumatism
— id: 70107, year: 2006, vol: 54, page: S163, stat: Journal Article,

Hypothyroidism and antithyroglobulin and antithyroperoxidase antibodies in the pathogenesis of autoimmune associated congenital heart block
Askanase, Anca Dinu; Iloh, Ijeoma; Buyon, Jill P
2006 Oct;33(10):2099-2099, Journal of rheumatology
— id: 73522, year: 2006, vol: 33, page: 2099, stat: Journal Article,

Autoimmunity as a result of escape from RNA surveillance
Bachmann, Michael P; Bartsch, Holger; Gross, Joanne K; Maier, Shannon M; Gross, Timothy F; Workman, Jennifer L; James, Judith A; Farris, A Darise; Jung, Bettina; Franke, Claudia; Conrad, Karsten; Schmitz, Marc; Buttner, Cordula; Buyon, Jill P; Semsei, Imre; Harley, John B; Rieber, E Peter
2006 Aug 1;177(3):1698-1707, Journal of immunology
In previous studies, we detected a frame shift mutation in the gene encoding the autoantigen La of a patient with systemic lupus erythematosus. The mutant La mRNA contains a premature termination codon. mRNAs that prematurely terminate translation should be eliminated by RNA quality control mechanisms. As we find Abs specific for the mutant La form in approximately 30% of sera from anti-La-positive patients, we expected that mutant La mRNAs circumvent RNA control and the expression of mutant La protein could become harmful. Indeed, real-time PCR, immunostaining, and immunoblotting data of mice transgenic for the mutant La form show that mutant La mRNAs are not repressed in these animals and are translated to mutant La protein. In addition to the mutant La protein, we detected a minor portion of native human La in the mutant La-transgenic mice. Therefore, ribosomal frame shifting may allow the mutant La mRNA to escape from RNA control. Interestingly, expression of the mutant La mRNA results in a lupus-like disease in the experimental mice. Consequently, escape of mutant La mRNA from RNA control can have two effects: it 1) results in the expression of an immunogenic (neo)epitope, and 2) predisposes to autoimmunity
— id: 73523, year: 2006, vol: 177, page: 1698, stat: Journal Article,

Impaired clearance of apoptotic cardiocytes and the pathogenesis of congenital heart block (CHB): Link to Anti-SSA/Ro-SSB/La antibodies
Clancy, RM; Neufing, PJ; O'Mahony, M; Zheng, P; Llanos, C; Gordon, TP; Buyon, JP
2006 SEP ;54(9):S551-S551, Arthritis & rheumatism
— id: 70126, year: 2006, vol: 54, page: S551, stat: Journal Article,

Impaired clearance of apoptotic cardiocytes is linked to anti-SSA/Ro and -SSB/La antibodies in the pathogenesis of congenital heart block
Clancy, Robert M; Neufing, Petra J; Zheng, Ping; O'Mahony, Marguerita; Nimmerjahn, Falk; Gordon, Tom P; Buyon, Jill P
2006 Sep;116(9):2413-2422, Journal of clinical investigation
The role of cardiocytes in physiologic removal of apoptotic cells and the subsequent effect of surface binding by anti-SSA/Ro and -SSB/La antibodies was addressed. Initial experiments evaluated induction of apoptosis by extrinsic and intrinsic pathways. Nuclear injury and the translocation of SSA/Ro and SSB/La antigens to the fetal cardiocyte plasma membrane were common downstream events of Fas and TNF receptor ligation, requiring caspase activation. As assessed by phase-contrast and confirmed by confocal microscopy, coculturing of healthy cardiocytes with cardiocytes rendered apoptotic via extrinsic pathways revealed a clearance mechanism that to our knowledge has not yet been described. Cultured fetal cardiocytes expressed phosphatidylserine receptors (PSRs), as did cardiac tissue from a fetus with congenital heart block (CHB) and an age-matched control. Phagocytic uptake was blocked by anti-PSR antibodies and was significantly inhibited following preincubation of apoptotic cardiocytes with chicken and murine anti-SSA/Ro and -SSB/La antibodies, with IgG from an anti-SSA/Ro- and -SSB/La-positive mother of a CHB child, but not with anti-HLA class I antibody. In a murine model, anti-Ro60 bound and inhibited uptake of apoptotic cardiocytes from wild-type but not Ro60-knockout mice. Results suggest that resident cardiocytes participate in physiologic clearance of apoptotic cardiocytes but that clearance is inhibited by opsonization via maternal autoantibodies, resulting in accumulation of apoptotic cells, promoting inflammation and subsequent scarring
— id: 67279, year: 2006, vol: 116, page: 2413, stat: Journal Article,

Utility of cardiac monitoring in fetuses at risk of congenital heart block: The PR interval and dexamethasone evaluation (PRIDE) prospective study
Friedman, DM; Kim, MY; Copel, JA; Davis, C; Buyon, JP
2006 SEP ;54(9):S689-S689, Arthritis & rheumatism
— id: 70130, year: 2006, vol: 54, page: S689, stat: Journal Article,

TGF beta, a strong fetal genetic candidate, in the development of congenital heart block (CHB)
Izmirly, PM; Harris, RR; Merrill, JT; Harley, JB; Backer, C; Clancy, RM; Buyon, JP
2006 SEP ;54(9):S536-S537, Arthritis & rheumatism
— id: 70124, year: 2006, vol: 54, page: S536, stat: Journal Article,

Antibody cross-reactivity of ScFv fragments that specifically recognize human apoptotic fetal cardiocytes and recombinant human Ro60
Llanos, C; Buyon, JP; Bennett, S; Lavner, L; Clancy, RM
2006 SEP ;54(9):S67-S68, Arthritis & rheumatism
— id: 70101, year: 2006, vol: 54, page: S67, stat: Journal Article,

Antibodies to two regulators of coagulation and complement are prevalent in SLE even in the absence conventional antiphospholipid antibody tests
O'Brien, K; Kamp, S; Wilson, S; Hutcheson, J; Kamp, P; Sigler, L; Petri, M; Buyon, JP; Merrill, JT
2006 SEP ;54(9):S262-S262, Arthritis & rheumatism
— id: 70112, year: 2006, vol: 54, page: S262, stat: Journal Article,

Beneficial effects of hormone therapy on health related quality of life (HRQOL) in systemic lupus erythematosus (SLE): Results of SELENA OCP and HRT randomized controlled trials (RCTS)
Petri, M; Buyon, J; Sigler, L; Qazi, U; Kim, M; Palmer, S; Crawford, B; Strand, V
2006 SEP ;54(9):S447-S447, Arthritis & rheumatism
— id: 70121, year: 2006, vol: 54, page: S447, stat: Journal Article,

Contribution of vasculopathy to lupus nephritis: Endothelial protein C receptor levels and genotype
Rivera, TL; Izmirly, PM; Buyon, JP; Clancy, RM
2006 SEP ;54(9):S824-S825, Arthritis & rheumatism
— id: 70137, year: 2006, vol: 54, page: S824, stat: Journal Article,

Inadequate anti-idiotypic response, as assessed by antibodies to La/SSB complementary epitopes: A novel component in the development of neonatal lupus
Stea, EA; Routsias, JG; Clancy, RM; Buyon, JP; Moutsopoulos, HM; Tzioufas, AG
2006 APR ;65(4):A18-A18, Annals of rheumatic diseases
— id: 74152, year: 2006, vol: 65, page: A18, stat: Journal Article,

Anti-La/SSB antiidiotypic antibodies in maternal serum: a marker of low risk for neonatal lupus in an offspring
Stea, Eleni A; Routsias, John G; Clancy, Robert M; Buyon, Jill P; Moutsopoulos, Haralampos M; Tzioufas, Athanasios G
2006 Jul;54(7):2228-2234, Arthritis & rheumatism
OBJECTIVE: The anti-La/SSB response to major B cell epitopes of La/SSB can be blocked by an active idiotypic/antiidiotypic network, which can be identified using synthetic complementary epitopes deduced from the sequence of the major B cell epitopes of the molecule. This study evaluated the role of this network in pregnant women with anti-Ro/SSA and/or anti-La/SSB antibodies in the development of neonatal lupus syndrome (NLS). METHODS. Sixty-three serum samples collected from anti-Ro/anti-La-positive women during pregnancy or within 6 months after delivery were obtained from the Research Registry for Neonatal Lupus and the PR Interval Dexamethasone Evaluation study. These samples, as well as 30 sera from healthy individuals, were tested in a blinded manner by enzyme-linked immunosorbent assay against synthetic peptides corresponding to major B cell epitopes and complementary epitopes of La/SSB. RESULTS. Sera from mothers giving birth to a healthy child and having no history of a child with NLS exhibited higher antiidiotypic antibody activity compared with mothers carrying a child with NLS (P < 0.0001) or mothers giving birth to a healthy child but who previously gave birth to a child with NLS (P = 0.0151). Sera from mothers of healthy children, which exhibited no apparent epitope activity against amino acids 349-364, revealed a significantly greater frequency of hidden anti-349-364aa epitope responses, blocked by antiidiotypic antibodies, as compared with sera from women pregnant with an affected child (P = 0.0094). CONCLUSION: The presence of antiidiotypic antibodies to autoantibodies against La/SSB may protect the fetus by blocking pathogenic maternal autoantibodies. Testing for these antiidiotypic responses may be useful in predicting a decreased risk of NLS
— id: 67280, year: 2006, vol: 54, page: 2228, stat: Journal Article,

The effect of moderate-dose corticosteroids in preventing severe flares in patients with serologically active, but clinically stable, systemic lupus erythematosus: Findings of a prospective, randomized, double-blind, placebo-controlled trial
Tseng, Chung-E; Buyon, Jill P; Kim, Mimi; Belmont, H Michael; Mackay, Meggan; Diamond, Betty; Marder, Galina; Rosenthal, Pamela; Haines, Kathleen; Ilie, Virginia; Abramson, Steven B
2006 Oct 30;54(11):3623-3632, Arthritis & rheumatism
OBJECTIVE: Serial measurements of anti-double-stranded DNA (anti-dsDNA) and complement are routine in the management of systemic lupus erythematosus (SLE), but their utility as biomarkers in preemptive treatment to prevent flares remains a subject of controversy. We hypothesized that concomitant elevation of anti-dsDNA and C3a can predict SLE activity in patients with stable or inactive disease and that short-term treatment with corticosteroids can avert flares. METHODS: In this prospective, randomized, double-blind, placebo-controlled trial, 154 patients were evaluated monthly for up to 18 months, with measurements of C3a, C3, C4, CH50, and anti-dsDNA levels. Patients who remained clinically stable but showed serologic evidence of an SLE flare (elevation of both the anti-dsDNA level by 25% and the C3a level by 50% over the previous 1-2 monthly visits) were randomized to receive either prednisone or placebo therapy at a dosage of 30 mg/day for 2 weeks, 20 mg/day for 1 week, and 10 mg/day for 1 week. RESULTS: Forty-one patients (21 randomized to prednisone and 20 randomized to placebo) experienced a serologic flare. Analysis of severe flares occurring </=90 days from randomization revealed that 6 occurred in patients taking placebo and none occurred in patients taking prednisone (P = 0.007). Severe flares resulted in an increase in the prednisone dosage to >40 mg/day and/or the addition of an immunosuppressive agent. Furthermore, improvement in scores on the Systemic Lupus Erythematosus Disease Activity Index, decreased levels of anti-dsDNA antibodies, and increased levels of C4 occurred 1 month after initiation of prednisone treatment. CONCLUSION: These preliminary data support our hypothesis that in a subset of clinically stable SLE patients with a combination of elevated C3a and anti-dsDNA levels, short-term corticosteroid therapy may avert a severe flare
— id: 69280, year: 2006, vol: 54, page: 3623, stat: Journal Article,

Frequency of neuropsychiatric disorders, autoimmune diseases and autoantibodies in anti-SSA/Ro-exposed children
Askanase, AD; Katholi, M; Buyon, JP
2005 SEP ;52(9):S528-S528, Arthritis & rheumatism
— id: 59290, year: 2005, vol: 52, page: S528, stat: Journal Article,

Prospective evaluation of disease progression in mothers of children enrolled in the research registry for neonatal lupus
Birnbaum, BK; Katholi, M; Buyon, JP
2005 SEP ;52(9):S464-S464, Arthritis & rheumatism
— id: 59286, year: 2005, vol: 52, page: S464, stat: Journal Article,

Circulating endothelial cells. Biomarker of vascular disease
Blann, Andrew D; Woywodt, Alexander; Bertolini, Francesco; Bull, Todd M; Buyon, Jill P; Clancy, Robert M; Haubitz, Marion; Hebbel, Robert P; Lip, Gregory Y H; Mancuso, Patrizia; Sampol, Jose; Solovey, Anna; Dignat-George, Francoise
2005 Feb;93(2):228-235, Thrombosis & haemostasis
Recent research has recognised new populations of non-hematopoietic cells in the blood. One of these, circulating endothelial cells (CECs), often defined by the expression of membrane glycoprotein CD146, are rarely found in the blood in health, but raised numbers are present in a wide variety of human conditions, including inflammatory, immune, infectious, neoplastic and cardiovascular disease, and seem likely to be evidence of profound vascular insult. An additional population are endothelial progenitor cells, defined by the co-expression of endothelial and immaturity cell surface molecules and also by the ability to form colonies in vitro. Although increased numbers of CECs correlate with other markers of vascular disease, questions remain regarding the precise definition, cell biology and origin of CECs. For example, they may be damaged, necrotic or apopototic, or alive, and could possess procoagulant and/or proinflammatory properties. However, since these cells seem to be representative of in situ endothelium, their phenotype may provide useful information. Indeed, whatever their phenotype, there is growing evidence that CECs may well be a novel biomarker, the measurement of which will have utility in various clinical settings related to vascular injury. Despite this promise, progress is impeded by the diversity of methodologies used to detect these cells. Accordingly, results are sometimes inconclusive and even conflicting. Nevertheless, increased CECs predict adverse cardiovascular events in acute coronary syndromes, suggesting they may move from being simply a research index to having a role in the clinic. The objective of the present communication is to condense existing data on CECs, briefly compare them with progenitor cells, and summarise possible mechanism(s) by which they may contribute to vascular pathology
— id: 73527, year: 2005, vol: 93, page: 228, stat: Journal Article,

Dispelling the preconceived notion that lupus pregnancies result in poor outcomes
Buyon, Jill P
2005 Sep;32(9):1641-1642, Journal of rheumatology
— id: 73525, year: 2005, vol: 32, page: 1641, stat: Journal Article,

Identifying an early marker for congenital heart block: when is a long PR interval too long? Comment on the article by Sonesson et al
Buyon, Jill P; Askanase, Anca Dinu; Kim, Mimi Y; Copel, Joshua A; Friedman, Deborah M
2005 Apr;52(4):1341-1342, Arthritis & rheumatism
— id: 73526, year: 2005, vol: 52, page: 1341, stat: Journal Article,

Autoantibody-associated congenital heart block: TGFbeta and the road to scar
Buyon, Jill P; Clancy, Robert M
2005 Jan;4(1):1-7, Autoimmunity Reviews
Few diseases exemplify the integration of research from bench to bedside as well as neonatal lupus (NL), often referred to as a model of passively acquired autoimmunity. The signature histologic lesion of autoimmune congenital heart block (CHB) is fibrosis of the conducting tissue and, in some cases, the surrounding myocardium. It is astounding how rapid, and in most cases, irreversible, the fibrotic response to injury is. The mechanism by which maternal anti-SSA/Ro-SSB/La antibodies initiate and finally eventuate in atrioventricular (AV) nodal scarring is not yet defined. In vitro and in vivo studies suggest that one pathologic cascade leading to scarring may be initiated via apoptosis, resulting in the translocation of SSA/Ro-SSB/La antigens and surface binding by maternal autoantibodies. Subsequently, the Fc portion of the bound immunoglobulin engages Fcgamma receptors on tissue macrophages, resulting in the release of TGFbeta at a threshold that favors a pro-fibrotic milieu and irreversible scarring. This cascade also involves a tissue-specific activation of TGFbeta, which promotes the transdifferentiation of fibroblasts into myofibroblasts, a scarring phenotype. Phagocytosis of opsonized apoptotic cardiocytes is distinct from macrophage pathways engaged in physiologic clearance of dying tissue, which also results in the release of TGFbeta but in the latter case appropriately serves to dampen inflammation. Downregulation of TGFbeta (activation/secretion pathway) may provide the basis of a novel approach to treatment of CHB in the future
— id: 55784, year: 2005, vol: 4, page: 1, stat: Journal Article,

Neonatal lupus: basic research and clinical perspectives
Buyon, Jill P; Clancy, Robert M
2005 May;31(2):299-313, Rheumatic diseases clinics of North America
Neonatal lupus, although quite rare, carries a significant mortality and morbidity when the fetal heart is the targeted organ. Anti-SSA/Ro-SSB/La antibodies are present in more than 85% of mothers whose fetuses are identified with congenital heart block (CHB) in a structurally normal heart, but the risk for a woman who has the candidate antibodies to have a child who has CHB is approximately 2%. Although the precise pathogenic mechanism of injury remains unknown, it is clear that the antibodies alone are insufficient to cause disease, and fetal factors are likely contributory. In vivo and in vitro evidence supports a pathologic cascade, involving apoptosis of cardiocytes, surface translocation of Ro and La antigens, binding of maternal autoantibodies, secretion of profibrosing factors (eg, tumor growth factor ) from the scavenging macrophages, and modulation of cardiac fibroblasts to a myofibroblast-scarring phenotype. The spectrum of cardiac abnormalities encompasses varying degrees of block identified in utero and late-onset cardiomyopathy. Better echocardiographic measurements that identify first-degree block in utero may be the optimal approach for pregnant women at risk; prophylactic therapies, including treatment with intravenous immunoglobulin, await larger trials
— id: 55992, year: 2005, vol: 31, page: 299, stat: Journal Article,

The effect of combined estrogen and progesterone hormone replacement therapy on disease activity in systemic lupus erythematosus: a randomized trial
Buyon, Jill P; Petri, Michelle A; Kim, Mimi Y; Kalunian, Kenneth C; Grossman, Jennifer; Hahn, Bevra H; Merrill, Joan T; Sammaritano, Lisa; Lockshin, Michael; Alarcon, Graciela S; Manzi, Susan; Belmont, H Michael; Askanase, Anca D; Sigler, Lisa; Dooley, Mary Anne; Von Feldt, Joan; McCune, W Joseph; Friedman, Alan; Wachs, Jane; Cronin, Mary; Hearth-Holmes, Michelene; Tan, Mark; Licciardi, Frederick
2005 Jun 21;142(12 Pt 1):953-962, Annals of internal medicine
BACKGROUND: There is concern that exogenous female hormones may worsen disease activity in women with systemic lupus erythematosus (SLE). OBJECTIVE: To evaluate the effect of hormone replacement therapy (HRT) on disease activity in postmenopausal women with SLE. DESIGN: Randomized, double-blind, placebo-controlled noninferiority trial conducted from March 1996 to June 2002. SETTING: 16 university-affiliated rheumatology clinics or practices in 11 U.S. states. PATIENTS: 351 menopausal patients (mean age, 50 years) with inactive (81.5%) or stable-active (18.5%) SLE. Interventions: 12 months of treatment with active drug (0.625 mg of conjugated estrogen daily, plus 5 mg of medroxyprogesterone for 12 days per month) or placebo. The 12-month follow-up rate was 82% for the HRT group and 87% for the placebo group. MEASUREMENTS: The primary end point was occurrence of a severe flare as defined by Safety of Estrogens in Lupus Erythematosus, National Assessment-Systemic Lupus Erythematosus Disease Activity Index composite. RESULTS: Severe flare was rare in both treatment groups: The 12-month severe flare rate was 0.081 for the HRT group and 0.049 for the placebo group, yielding an estimated difference of 0.033 (P = 0.23). The upper limit of the 1-sided 95% CI for the treatment difference was 0.078, within the prespecified margin of 9% for noninferiority. Mild to moderate flares were significantly increased in the HRT group: 1.14 flares/person-year for HRT and 0.86 flare/person-year for placebo (relative risk, 1.34; P = 0.01). The probability of any type of flare by 12 months was 0.64 for the HRT group and 0.51 for the placebo group (P = 0.01). In the HRT group, there were 1 death, 1 stroke, 2 cases of deep venous thrombosis, and 1 case of thrombosis in an arteriovenous graft; in the placebo group, 1 patient developed deep venous thrombosis. LIMITATIONS: Findings are not generalizable to women with high-titer anticardiolipin antibodies, lupus anticoagulant, or previous thrombosis. CONCLUSIONS: Adding a short course of HRT is associated with a small risk for increasing the natural flare rate of lupus. Most of these flares are mild to moderate. The benefits of HRT can be balanced against the risk for flare because HRT did not significantly increase the risk for severe flare compared with placebo
— id: 55990, year: 2005, vol: 142, page: 953, stat: Journal Article,

Reply
Clancy RM; Buyon JP
2005 May;52(5):1625-1626, Arthritis & rheumatism
— id: 55993, year: 2005, vol: 52, page: 1625, stat: Journal Article,

Structure/function of apoptotic pathways and subsequent macrophage activation link anti-Ro/La antibodies to the pathogenesis of congenital heart block
Clancy, R; Zheng, P; Buyon, JP
2005 SEP ;52(9):S449-S449, Arthritis & rheumatism
— id: 59283, year: 2005, vol: 52, page: S449, stat: Journal Article,

Hypoxia, an environmental stress, in initiation and progression of the pathologic cascade to congenital heart block
Clancy, R; Zheng, P; Gardner, L; Buyon, JP
2005 SEP ;52(9):S303-S303, Arthritis & rheumatism
— id: 59276, year: 2005, vol: 52, page: S303, stat: Journal Article,

Maternal antibody responses to the 52-kd SSA/RO p200 peptide and the development of fetal conduction defects
Clancy, Robert M; Buyon, Jill P; Ikeda, Keigo; Nozawa, Kazuhisa; Argyle, Dionne A; Friedman, Deborah M; Chan, Edward K L
2005 Oct;52(10):3079-3086, Arthritis & rheumatism
OBJECTIVE: To identify a finer level of antibody specificity for risk of congenital heart block (CHB) than reactivity to 52-kd SSA/Ro (Ro 52). METHODS: Serum from mothers enrolled in the Research Registry for Neonatal Lupus and the observational PR Interval and Dexamethasone Evaluation (PRIDE) study was evaluated for reactivity against peptide aa200-239 of Ro 52 (p200), recently reported to be associated with a higher risk of CHB. RESULTS: The majority of 156 Ro 52-positive sera tested were reactive with p200 (>3 SD above control), irrespective of the clinical status of the child. Optical density (OD) values of p200 did not differ significantly among mothers of children with CHB (mean +/- SD 0.187 +/- 0.363), mothers of children with rash (mean +/- SD 0.176 +/- 0.356), and mothers of children without neonatal lupus (mean +/- SD 0.229 +/- 0.315). Reactivity against p200 was found in 80 of 104 mothers of children with CHB (77%), 24 of 30 mothers of children with rash (80%), and 21 of 22 mothers who delivered healthy children and had no children with neonatal lupus (95%) (P not significant for all comparisons). Sera from 4 mothers of children with CHB with varied p200 titers (OD range 0.025-1.818) bound to the surface of non-permeabilized apoptotic, but not proliferating, human fetal cardiocytes. In 32 Ro 52-positive women who completed the PRIDE study (22 with no child with neonatal lupus, 7 with a child with CHB, and 3 with a child with rash) in whom p200 levels were determined during pregnancy, the correlation between level of p200 (OD range 0.000-1.170) and maximal fetal PR interval (range 115-168 msec) was not significant (rho = 0.107, P = 0.58). CONCLUSION: Reactivity to p200 is a dominant but not uniform anti-Ro 52 response in women whose children have CHB. Since exposure to this antibody specificity was observed with a similar frequency in children without CHB born to mothers with anti-Ro 52, additional factors are necessary to convert risk to disease expression
— id: 58889, year: 2005, vol: 52, page: 3079, stat: Journal Article,

Complete atrioventricular block diagnosed prenatally: anything new on the block?
Friedman, D M; Buyon, J P
2005 Jul;26(1):2-3, Ultrasound in obstetrics & gynecology
— id: 73544, year: 2005, vol: 26, page: 2, stat: Journal Article,

Circulating endothelial cells and soluble endothelial protein C receptor predict the presence of carotid plaque in minority SLE patients
Gehrie, ER; Reynolds, HR; Buyon, JP; Clancy, R
2005 SEP ;52(9):S606-S606, Arthritis & rheumatism
— id: 59293, year: 2005, vol: 52, page: S606, stat: Journal Article,

Mycophenolate mofetil or intravenous cyclophosphamide for lupus nephritis
Ginzler, Ellen M; Dooley, Mary Anne; Aranow, Cynthia; Kim, Mimi Y; Buyon, Jill; Merrill, Joan T; Petri, Michelle; Gilkeson, Gary S; Wallace, Daniel J; Weisman, Michael H; Appel, Gerald B
2005 Nov 24;353(21):2219-2228, New England journal of medicine
BACKGROUND: Since anecdotal series and small, prospective, controlled trials suggest that mycophenolate mofetil may be effective for treating lupus nephritis, larger trials are desirable. METHODS: We conducted a 24-week randomized, open-label, noninferiority trial comparing oral mycophenolate mofetil (initial dose, 1000 mg per day, increased to 3000 mg per day) with monthly intravenous cyclophosphamide (0.5 g per square meter of body-surface area, increased to 1.0 g per square meter) as induction therapy for active lupus nephritis. A change to the alternative regimen was allowed at 12 weeks in patients who did not have an early response. The study protocol specified adjunctive care and the use and tapering of corticosteroids. The primary end point was complete remission at 24 weeks (normalization of abnormal renal measurements and maintenance of baseline normal measurements). A secondary end point was partial remission at 24 weeks. RESULTS: Of 140 patients recruited, 71 were randomly assigned to receive mycophenolate mofetil and 69 were randomly assigned to receive cyclophosphamide. At 12 weeks, 56 patients receiving mycophenolate mofetil and 42 receiving cyclophosphamide had satisfactory early responses. In the intention-to-treat analysis, 16 of the 71 patients (22.5 percent) receiving mycophenolate mofetil and 4 of the 69 patients receiving cyclophosphamide (5.8 percent) had complete remission, for an absolute difference of 16.7 percentage points (95 percent confidence interval, 5.6 to 27.9 percentage points; P=0.005), meeting the prespecified criteria for noninferiority and demonstrating the superiority of mycophenolate mofetil to cyclophosphamide. Partial remission occurred in 21 of the 71 patients (29.6 percent) and 17 of the 69 patients (24.6 percent), respectively (P=0.51). Three patients assigned to cyclophosphamide died, two during protocol therapy. Fewer severe infections and hospitalizations but more diarrhea occurred among those receiving mycophenolate. CONCLUSIONS: In this 24-week trial, mycophenolate mofetil was more effective than intravenous cyclophosphamide in inducing remission of lupus nephritis and had a more favorable safety profile
— id: 73524, year: 2005, vol: 353, page: 2219, stat: Journal Article,

Autoantibodies against the serotoninergic 5-HT4 receptor and congenital heart block: a reassessment
Kamel, Rehab; Eftekhari, Pierre; Clancy, Robert; Buyon, Jill P; Hoebeke, Johan
2005 Aug;25(1):72-76, Journal of autoimmunity
The specificity of autoantibodies against the serotoninergic 5-HT4 receptor in congenital heart block has led to conflicting observations. In order to clarify the situation, a collaborative effort was undertaken to discover the reasons for these discrepancies and to reassess the importance of such autoantibodies by making use of the Research Registry for Neonatal Lupus. Sera from 128 patients (101 anti-SSA/Ro52 positive mothers among which 74 have children with congenital heart block (CHB), 9 anti-SSA/Ro52 negative patients of which 1 had a child with heart block and 18 healthy donors) were assessed in a single blind test using an ELISA coated with a 5-HT4 receptor-derived peptide. Discrepancies between previous observations in our two groups could be ascribed to small differences in the set up of the assay. Of the 75 sera from mothers of children with CHB, 12 were reactive with the 5-HT4 peptide. Four sera among which three were from 35 Ro52 negative mothers without affected children as well as 2 in the 18 controls were positive. Interestingly, in 1 mother with an isolated child with CHB but who had no detectable anti-SSA/Ro52 antibodies and 1 mother with a child with a structural heart block and no detectable antibodies to any component of SSA/Ro, reactivity with the 5-HT4 receptor was noted. While 5-HT4 receptor autoantibodies do not have the predictive value of anti-Ro52 autoantibodies, the presence of these antibodies in a minor subset of mothers whose children have CHB suggests an additional risk factor which may contribute to the pathogenesis of disease
— id: 67283, year: 2005, vol: 25, page: 72, stat: Journal Article,

The role of biomarkers in the assessment of lupus
Merrill, Joan T; Buyon, Jill P
2005 Oct;19(5):709-726, Bailliere's best practice & research. Clinical rheumatology
Although considered a prototypic autoimmune disease, the hallmark of systemic lupus erythematosus (SLE) is its heterogeneity. Accordingly, manifestations can vary widely from person to person, with the potential involvement of virtually any bodily organ. Furthermore, the genetic abnormalities underlying this condition are complicated, with diverse genetic polymorphisms described in different ethnic groups, strongly suggesting that the actual pathology underlying the immunologic disarray might not be the same for each patient. Evolving concepts of genetics and immunity have clarified that patients can carry unique arrays of exacerbating and protective factors. These factors, in conjunction with variable environmental triggers for SLE, probably determine the sequelae that an individual experiences. Therefore, it is not surprising that the clinical manifestations are diverse, the temporal sequence of organ involvement often unpredictable, and that the flares of inflammatory activity that characterize SLE can either remit without consequence or leave permanent damage in their wake. It is widely accepted that the current standard of care for SLE patients is inadequate. Programs to develop and test new drug and/or device therapies have been ongoing since the mid-1990s but have encountered formidable obstacles. With the current burst of drug discovery and the advent of several large international trials of promising new agents, the challenge to overcome these obstacles has never been greater. A burgeoning literature in the past decades nevertheless suggests that despite the complexities of the many immunologic pathways that impact on SLE, characteristic biologic markers are emerging as potential signposts that can characterize patient subgroups, predict prognosis, mark the exacerbations and remissions of SLE flares, and serve as endpoints in the determination of the dosing and timing of immune-modulating treatments. Several of the promising biomarkers are addressed in this chapter
— id: 61259, year: 2005, vol: 19, page: 709, stat: Journal Article,

Exposure and binding of selected immunodominant La/SSB epitopes on human apoptotic cells
Neufing, Petra J; Clancy, Robert M; Jackson, Michael W; Tran, Hai Bac; Buyon, Jill P; Gordon, Tom P
2005 Dec;52(12):3934-3942, Arthritis & rheumatism
OBJECTIVE: Opsonization of apoptotic cells by autoantibodies bound to surface membrane-translocated La/SSB antigens may initiate tissue damage in the setting of congenital heart block. By injecting pregnant mice with human anti-La antibodies, we previously demonstrated the formation of IgG-apoptotic cell complexes in the developing mouse fetus; however, the binding of anti-La antibodies to human-specific epitopes could not be addressed. Accordingly, the objective of the current study was to delineate the epitope specificity of human La antibodies that are exposed on the surface of apoptotic cells. METHODS: We used fluorescence microscopy and flow cytometry to assess the binding of human anti-La antibodies affinity purified against immunodominant epitopes of La to human cells undergoing spontaneous apoptosis, in a murine xenograft model in vivo and in cultured human fetal cardiocytes rendered apoptotic in vitro, respectively. RESULTS: Anti-La antibodies bound to immunodominant epitopes of La within the NH(2)-terminus and the RNA recognition motif (RRM) region of apoptotic human cells, in both xenografts and fetal cardiocytes. In contrast, human antibodies affinity purified against the COOH-terminal La epitope did not bind apoptotic cells in either model. This defines the topology of redistributed La during apoptosis, with surface exposure of the NH(2)-terminus and RRM regions. The potential importance of anti-La NH(2)-terminal and anti-La RRM specificity was confirmed by detection of this reactivity in mothers of children with congenital heart block. CONCLUSION: These findings provide insight into both the molecular modification of the La autoantigen during apoptosis and the specificity of antibodies capable of binding to surface-exposed La. Subsequent formation of surface immune complexes may lead to tissue injury in patients with autoimmune diseases such as congenital heart block
— id: 67282, year: 2005, vol: 52, page: 3934, stat: Journal Article,

Combined oral contraceptives in women with systemic lupus erythematosus
Petri, Michelle; Kim, Mimi Y; Kalunian, Kenneth C; Grossman, Jennifer; Hahn, Bevra H; Sammaritano, Lisa R; Lockshin, Michael; Merrill, Joan T; Belmont, H Michael; Askanase, Anca D; McCune, W Joseph; Hearth-Holmes, Michelene; Dooley, Mary Anne; Von Feldt, Joan; Friedman, Alan; Tan, Mark; Davis, John; Cronin, Mary; Diamond, Betty; Mackay, Meggan; Sigler, Lisa; Fillius, Michael; Rupel, Ann; Licciardi, Frederick; Buyon, Jill P
2005 Dec 15;353(24):2550-2558, New England journal of medicine
BACKGROUND: Oral contraceptives are rarely prescribed for women with systemic lupus erythematosus, because of concern about potential negative side effects. In this double-blind, randomized, noninferiority trial, we prospectively evaluated the effect of oral contraceptives on lupus activity in premenopausal women with systemic lupus erythematosus. METHODS: A total of 183 women with inactive (76 percent) or stable active (24 percent) systemic lupus erythematosus at 15 U.S. sites were randomly assigned to receive either oral contraceptives (triphasic ethinyl estradiol at a dose of 35 microg plus norethindrone at a dose of 0.5 to 1 mg for 12 cycles of 28 days each; 91 women) or placebo (92 women) and were evaluated at months 1, 2, 3, 6, 9, and 12. Subjects were excluded if they had moderate or high levels of anticardiolipin antibodies, lupus anticoagulant, or a history of thrombosis. RESULTS: The primary end point, a severe lupus flare, occurred in 7 of 91 subjects receiving oral contraceptives (7.7 percent) as compared with 7 of 92 subjects receiving placebo (7.6 percent). The 12-month rates of severe flare were similar: 0.084 for the group receiving oral contraceptives and 0.087 for the placebo group (P=0.95; upper limit of the one-sided 95 percent confidence interval for this difference, 0.069, which is within the prespecified 9 percent margin for noninferiority). Rates of mild or moderate flares were 1.40 flares per person-year for subjects receiving oral contraceptives and 1.44 flares per person-year for subjects receiving placebo (relative risk, 0.98; P=0.86). In the group that was randomized to receive oral contraceptives, there was one deep venous thrombosis and one clotted graft; in the placebo group, there was one deep venous thrombosis, one ocular thrombosis, one superficial thrombophlebitis, and one death (after cessation of the trial). CONCLUSIONS: Our study indicates that oral contraceptives do not increase the risk of flare among women with systemic lupus erythematosus whose disease is stable
— id: 62351, year: 2005, vol: 353, page: 2550, stat: Journal Article,

Shedding of endothelial protein C receptor contributes to vasculopathy and renal injury in lupus: In vivo and in vitro evidence
Sesin, Carlos A; Yin, Xiaoming; Esmon, Charles T; Buyon, Jill P; Clancy, Robert M
2005 Jul;68(1):110-120, Kidney international
Shedding of endothelial protein C receptor contributes to vasculopathy and renal injury in lupus: In vivo and in vitro evidence. Background. Candidate biomarkers for vasculopathy in systemic lupus erythematosus (SLE) include circulating endothelial cells and the recently identified endothelial protein C receptor (EPCR) which, when shed, promotes a thrombotic diathesis. This study sought correlation between plasma levels of soluble EPCR and disease manifestation/severity, with a focus on lupus nephritis. Methods. In 81 SLE patients (evaluated cross-sectionally and longitudinally) and 59 healthy controls, levels of soluble EPCR and soluble E-selectin were assessed by sandwich enzyme-linked immunosorbent assay (ELISA), circulating endothelial cells isolated by immunomagnetic separation, and EPCR gene polymorphisms determined. Mechanisms of vascular injury were addressed in vitro in human aortic endothelial cells (HAEC) cultured in the presence and absence of interferon-gamma (IFN-gamma). Results. The mean level of soluble EPCR was significantly higher in SLE patients (263 +/- 13 ng/mL) than controls (174 +/- 11 ng/mL) (P < 0.0001). Patients with active or past renal involvement had significantly higher mean soluble EPCR levels (306 +/- 21 ng/mL) (N= 40) than patients without nephritis (228 +/- 14 ng/mL) (N= 41) (P= 0.0033). Mean soluble EPCR correlated positively with serum creatinine (R= 0.3429, P < 0.0001). The prevalence of the enhanced-shedding EPCR polymorphism A6936G was higher in SLE (41%) (N= 27) than controls (7%) (N= 29) (P= 0.0039). Patient and control plasma were also interrogated for soluble E-selectin, a comparator plasma marker. The results suggest that soluble E-selectin and soluble EPCR are not equivalent end points of vasculopathy and endothelial perturbation in SLE. Although in SLE patients the absence or diminished expression of membrane EPCR on circulating endothelial cells varied, the rare circulating endothelial cells detected in controls invariably expressed membrane-bound EPCR. IFN-gamma-treated HAEC expressed less membrane-bound EPCR [133 relative fluorescence units (rfu)] than untreated HAEC (275 rfu); more soluble EPCR was detected in IFN-gamma-treated (1.1 ng/10(6) cells) than untreated HAEC (0.65 ng/10(6) cells) (P= 0.027). Conclusion. The results obtained from this cross-sectional/longitudinal study support the hypothesis that the vascular dysfunction characteristic of SLE may be related to a dramatically altered distribution of EPCR, both soluble and membrane-bound forms
— id: 55991, year: 2005, vol: 68, page: 110, stat: Journal Article,

Pharmaeogenetics, enzymatic phenotyping and metabolite monitoring to guide treatment with Imuran in SLE patients
Askanase, A; Tseng, C; Lein, DO; Smith, KM; Bernstein, L; Belmont, HM; Scidman, E; Buyon, J
2004 SEP ;50(9):S448-S448, Arthritis & rheumatism
— id: 49047, year: 2004, vol: 50, page: S448, stat: Journal Article,

Hypothyroidism, anti-thyroglobulin and anti-thyroperoxidase antibodies in the pathogenesis of autoimmune-associated congenital heart block (CHB)
Askanase, AD; Iloh, I; Buyon, JP
2004 SEP ;50(9):S365-S366, Arthritis & rheumatism
— id: 49043, year: 2004, vol: 50, page: S365, stat: Journal Article,

Neonatal lupus syndromes
Buyon, J P; Rupel, A; Clancy, R M
2004 ;13(9):705-712, Lupus
The neonatal lupus syndromes (NLS), while quite rare, carry significant mortality and morbidity in cases of cardiac manifestations. Although anti-SSA/Ro-SSB/La antibodies are detected in > 85% of mothers whose fetuses are identified with congenital heart block (CHB) in a structurally normal heart, when clinicians applied this testing to their pregnant patients, the risk for a woman with the candidate antibodies to have a child with CHB was at or below 1 in 50. While the precise pathogenic mechanism of antibody-mediated injury remains unknown, it is clear that the antibodies alone are insufficient to cause disease and fetal factors are likely contributory. In vivo and in vitro evidence supports a pathologic cascade involving apoptosis of cardiocytes, surface translocation of Ro and La antigens, binding of maternal autoantibodies, secretion of profibrosing factors (e.g., TGFbeta) from the scavenging macrophages and modulation of cardiac fibroblasts to a myofibroflast scarring phenotype. The spectrum of cardiac abnormalities continues to expand, with varying degrees of block identified in utero and reports of late onset cardiomyopathy (some of which display endocardial fibroelastosis). Moreover, there is now clear documentation that incomplete blocks (including those improving in utero with dexamethasone) can progress postnatally, despite the clearance of the maternal antibodies from the neonatal circulation. Better echocardiographic measurements which identify first degree block in utero may be the optimal means of approaching pregnant women at risk. Prophylactic therapies, including treatment with intravenous immunoglobulin, await larger trials. In order to achieve advances at both the bench and bedside, national research registries established in the US and Canada are critical
— id: 47776, year: 2004, vol: 13, page: 705, stat: Journal Article,

Autoimmune Associated CHB: maternal anti-SSA/Ro-SSB/La antibodies and apotosis of fetal cardiocytes
Buyon, JP
2004 MAR 23 ;18(4):A403-A403, FASEB journal
— id: 46597, year: 2004, vol: 18, page: A403, stat: Journal Article,

Neonatal lupus syndrome: Cause and management
Buyon, JP; Clancy, RM
2004 JUL ;63(3):4-4, Annals of rheumatic diseases
— id: 49056, year: 2004, vol: 63, page: 4, stat: Journal Article,

Evaluation of Ro52 peptide antibody profiles in a large cohort of anti-Ro plus women whose children have neonatal lupus (NL) or are healthy
Clancy, RM; Nozawa, K; Friedman, D; Buyon, J; Chan, EK
2004 SEP ;50(9):S532-S532, Arthritis & rheumatism
— id: 49050, year: 2004, vol: 50, page: S532, stat: Journal Article,

Antibody cross-reactivity of scFv fragments that specifically recognize human apoptotic fetal cardiocytes with recombinant human ro52
Clancy, RM; Yin, XM; Buyon, J
2004 SEP ;50(9):S209-S209, Arthritis & rheumatism
— id: 49031, year: 2004, vol: 50, page: S209, stat: Journal Article,

Genetic association of cutaneous neonatal lupus with HLA class II and tumor necrosis factor alpha: implications for pathogenesis
Clancy, Robert M; Backer, Chelsea B; Yin, Xiaoming; Chang, Mary Wu; Cohen, Steven R; Lee, Lela A; Buyon, Jill P
2004 Sep;50(8):2598-2603, Arthritis & rheumatism
OBJECTIVE: Cutaneous neonatal lupus resembles subacute cutaneous lupus erythematosus (SCLE), and photosensitivity is a common symptom. Tumor necrosis factor alpha (TNFalpha) release by ultraviolet light-exposed keratinocytes may be exaggerated in SCLE patients who have the haplotype TNFalpha -308A;DRB1*03. Accordingly, this study was undertaken to seek genetic and histologic evidence for a role of TNFalpha in the pathogenesis of cutaneous neonatal lupus. METHODS: DNA was isolated from 83 children (22 with rash, 35 with congenital heart block [CHB], 26 unaffected siblings) and 58 mothers from the Research Registry for Neonatal Lupus. RESULTS: The -308A allele (associated with higher TNFalpha production), HLA-DRQB1*02, and HLA-DRB1*03 were each present in the majority of children with rash (64%, 68%, and 64%, respectively). The frequency of all 3 6p alleles occurring together in 1 individual was greater in children with rash than in children who had either CHB or no manifestation of neonatal lupus (59% versus 30%; P = 0.02). This association with neonatal lupus rash was equivalent to published findings in a cohort of patients with SCLE, but significantly greater than the association in patients with discoid lupus erythematosus. Prominent TNFalpha staining in the epidermis was observed in lesional skin from 3 children with rash, but not in skin from a healthy neonate. CONCLUSION: Taken together, the finding of a genetic predisposition to generate increased levels of TNFalpha following tissue injury and the histologic demonstration of TNFalpha in the target organ support the notion that this inflammatory cytokine plays a role in the pathogenesis of cutaneous neonatal lupus. Furthermore, the results of these studies provide evidence of a biologic link between neonatal lupus and the rash of SCLE
— id: 46137, year: 2004, vol: 50, page: 2598, stat: Journal Article,

Autoimmune-associated congenital heart block: dissecting the cascade from immunologic insult to relentless fibrosis
Clancy, Robert M; Buyon, Jill P
2004 Oct;280(2):1027-1035, Anatomical record. Part A, Discoveries in molecular, cellular, & evolutionary biology
Few diseases exemplify the integration of research from bench to bedside as well as neonatal lupus, often described as a model of passively acquired autoimmunity. The signature histologic lesion of autoimmune congenital heart block (CHB) is fibrosis of the conducting tissue and, in some cases, the surrounding myocardium. Although anti-SSA/Ro-SSB/La antibodies are detected in > 85% of mothers whose fetuses are identified with conduction abnormalities in a structurally normal heart, the risk for a woman with the candidate antibodies to have a child with CHB is 2%. The mechanism by which maternal anti-SSA/Ro-SSB/La antibodies initiate and finally eventuate in atrioventricular nodal scarring is not yet defined, but it is clear that the antibodies alone are insufficient to cause disease and fetal factors are likely contributory. Previous in vitro and in vivo studies suggest that the pathologic cascade is initiated via apoptosis, resulting in translocation of SSA/Ro-SSB/La antigens to the cell surface where they are bound by maternal autoantibodies. Subsequently, the Fc portion of the bound immunoglobulin engages Fcgamma receptors on tissue macrophages, resulting in release of TGF-beta at a threshold favoring a profibrotic milieu and irreversible scarring. This cascade also involves tissue-specific activation of TGF-beta, which promotes the modulation of fibroblasts into myofibroblasts, a scarring phenotype. Recent findings point to genetic polymorphisms that promote high production of TGF-beta as possible fetal risk factors for CHB. Further elucidation of maternal and fetal contributory factors should provide insight into the pathogenesis of CHB and the rarity of irreversible injury
— id: 48075, year: 2004, vol: 280, page: 1027, stat: Journal Article,

More to death than dying: apoptosis in the pathogenesis of SSA/Ro-SSB/La-associated congenital heart block
Clancy, Robert M; Buyon, Jill P
2004 Sep;30(3):589-602, x, Rheumatic diseases clinics of North America
The mechanism by which maternal anti-SSA/Ro-SSB/La antibodies initiate and perpetuate inflammation and eventuate in scarring of the atrioventricular node (the signature lesion of congenital heart block) is not yet defined. In vitro and in vivo studies suggest that one pathologic cascade that leads to scarring may be initiated by way of apoptosis which results in translocation of SSA/Ro-SSB/La antigens and subsequent surface binding by maternal autoantibodies. These opsonized cardiocytes are phagocytosed by macrophages,which secrete factors that modulate fibroblasts into myofibroblasts,a scarring phenotype. The exaggerated apoptosis (amplified physiologic apoptosis or defective clearance) that is observed in autopsy slides from fetuses who had congenital heart block may provide the pivotal clue to understanding the pathogenicity of the maternal autoantibodies
— id: 46167, year: 2004, vol: 30, page: 589, stat: Journal Article,

Immunohistologic evidence supports apoptosis, IgG deposition, and novel macrophage/fibroblast crosstalk in the pathologic cascade leading to congenital heart block
Clancy, Robert M; Kapur, Raj P; Molad, Yair; Askanase, Anca Dinu; Buyon, Jill P
2004 Jan;50(1):173-182, Arthritis & rheumatism
OBJECTIVE: To assess in vivo the pathologic cascade leading to fibrosis in congenital heart block (CHB). In vitro studies suggest that CHB is initiated via apoptosis, resulting in translocation of SSA/Ro and SSB/La antigens and surface binding by maternal autoantibodies. These opsonized cardiocytes are phagocytosed by macrophages, which secrete factors inducing fibrosis. METHODS: Immunohistochemistry analysis was performed on formalin-fixed sections of 4 fetal hearts identified in utero as having CHB or isolated myocarditis; mothers had anti-SSA/Ro and anti-SSB/La antibodies. RESULTS: Apoptosis was most extensive in fetuses dying early and most pronounced in regions containing conduction tissue. Deposition of IgG was observed in hearts from fetuses with CHB/myocarditis, but not in 3 control hearts, and was colocalized with apoptotic cells. Giant cells and macrophages (frequently seen proximal to IgG and apoptotic cells) were present in septal and thickened fibrous subendocardial regions, most apparent in the youngest fetuses. Septal tissue also revealed extensive areas of fibrosis and microcalcification in which a predominant smooth muscle actin (SMA)-positive infiltrate (myofibroblast scarring phenotype) was observed. In contrast, there were no macrophages or SMA-positive cells (other than those lining blood vessels) in septal tissue from control hearts, although rare macrophages were seen in the working myocardium. CONCLUSION: In summary, findings in this unique autopsy material paralleled those in in vitro studies. These data support the notion of exaggerated apoptosis, probably due to ongoing inflammation caused by IgG binding and ingestion by macrophages. Transdifferentiation of cardiac fibroblasts to a scarring phenotype may be a pathologic process initiated by maternal antibodies, and persistence of this phenotype even after birth may relate to the progression of block seen in some infants postpartum
— id: 42614, year: 2004, vol: 50, page: 173, stat: Journal Article,

The fetal Doppler mechanical PR interval: a validation study
Glickstein, Julie; Buyon, Jill; Kim, Mimi; Friedman, Deborah
2004 Jan-Feb;19(1):31-34, Fetal diagnosis & therapy
OBJECTIVE: To evaluate the accuracy of pulsed Doppler-derived fetal PR interval measurements obtained by physicians participating in a multicenter prospective fetal echocardiographic study. METHODS: Echocardiograms on healthy fetuses were performed and evaluated by 15 pediatric cardiologists/perinatologists across the United States who are participating in a larger clinical trial involving fetuses at risk for autoantibody-associated congenital heart block. Prior to enrolling women in the main trial, each physician was provided with a teaching tape to demonstrate how the pulsed Doppler-derived PR interval is measured. The procedure involves placing a gated pulsed Doppler sample volume in the left ventricle at the junction of the anterior leaflet of the mitral valve and the left ventricular outflow tract in an apical 5-chamber view, and simultaneously obtaining left ventricular filling and emptying. Time intervals are measured from the onset of the mitral A wave (atrial systole) to the onset of the aortic pulsed Doppler tracing (ventricular systole). This represents the mechanical PR interval. Each physician measured the pulsed Doppler-derived fetal PR interval on 5 different subjects recruited from the physician's specific site. To validate each physician's technique, the tapes were sent to a central facility and the same intervals were remeasured by an experienced central reader (D.M.F.). A physician was determined to have adequate ability to measure the fetal PR interval if all 5 measurements were within +/- 30 ms of the central reader's measurements, where 30 ms corresponds to 25% of the mean observed in normative PR interval data. This difference was deemed to be the minimum clinically important difference in Doppler PR interval. RESULTS: Fourteen of the 15 physicians were considered to have adequate ability to measure the fetal PR interval according to our established criterion. The overall mean difference between the physicians and the central reader's measurements was -0.26 +/- 11.04 ms (p = 0.84). In addition, 95% of the observed differences were included in the interval (-22.23 to 21.81), which is well within our clinically acceptable range of +/- 30 ms. CONCLUSIONS: The pulsed Doppler assessment of the mechanical PR interval in the fetus can be accurately performed after minimal training. This technique may be a valuable tool for identification of early and potentially reversible conduction abnormalities in fetuses at risk for more advanced and permanent forms of heart block associated with maternal antibodies to SSA/Ro-SSB/La
— id: 73532, year: 2004, vol: 19, page: 31, stat: Journal Article,

Anti-52 kDa Ro, anti-60 kDa Ro, and anti-La antibody profiles in neonatal lupus
Gordon, Patrick; Khamashta, Munther A; Rosenthal, Eric; Simpson, John M; Sharland, Gurleen; Brucato, Antonio; Franceschini, Franco; De Bosschere, Katrien; Meheus, Lydie; Meroni, Pier Luigi; Hughes, Graham R V; Buyon, Jill
2004 Dec;31(12):2480-2487, Journal of rheumatology
OBJECTIVE: Studies suggest that anti-52 kDa Ro antibodies are more sensitive and specific than anti-60 kDa Ro antibodies for neonatal lupus. However, these studies mainly used immunoblot or ELISA using recombinant protein, which have poor sensitivity for anti-60 kDa Ro antibodies. In addition, the control patients were not disease matched. We reassessed the sensitivity and specificity of anti-52 kDa Ro, anti-60 kDa Ro, and anti-La, addressing these limitations. METHODS AND RESULTS: To assess sensitivity, 125 mothers of children with neonatal lupus (NLM) were recruited. All maternal sera were assessed using a commercial line immunoassay that uses natural 60 kDa Ro protein (Inno-Lia ANA Update, Innogenetics NV, Gent, Belgium). By this method, 96% of the sera had antibodies to 60 kDa Ro, 86% to 52 kDa Ro, and 78% to 48 kDa La. Immunoblot of 65 NLM showed significantly fewer positive results for anti-60 kDa Ro (p < 0.001) and anti-52 kDa Ro (p < 0.05). Sensitivity of the 3 antibodies was assessed in the symptomatic mothers of children with congenital heart block (CHB) (78 women) and disease matched controls with unaffected children (65 women) using Inno-Lia ANA Update. The sensitivity of each antibody was compared by multiple logistic regression to adjust for maternal disease. There was no significant difference between the groups for 60 kDa Ro or for anti-52 kDa Ro antibody. However, there was a significant difference for the anti-La antibody (p = 0.001), with an odds ratio of 3.59. This translates to an increase in risk from a published 2% for CHB in an anti-Ro-positive mother to 3.1% if the woman is also anti-La antibody-positive, and to a decrease in risk to 0.9% if anti-La-negative. CONCLUSION: Contrary to previous reports, 52 kDa Ro as detected by Inno-Lia ANA Update is not more specific for or frequent in CHB than 60 kDa Ro. However, the presence of anti-La antibodies significantly increases the risk for CHB
— id: 73529, year: 2004, vol: 31, page: 2480, stat: Journal Article,

Challenges in bringing the bench to bedside in drug development for SLE
Merrill, Joan T; Erkan, Doruk; Buyon, Jill P
2004 Dec;3(12):1036-1046, Nature reviews. Drug discovery
It is now widely accepted that the current standard of care for systemic lupus erythematosus (SLE) patients is inadequate. There has not been a new medication approved for this disease in thirty years. Attempts to develop and test new drugs have been ongoing since the mid-1990s, but have encountered formidable obstacles. Current models for lupus pathogenesis have provided a theoretical framework for understanding how heterogeneous genetic defects might combine in various ways to increase susceptibility to SLE in different individuals, and could have important implications for new drug development. With the current burst of drug discovery and increased public awareness of SLE, the impetus to overcome these obstacles has never been greater
— id: 73528, year: 2004, vol: 3, page: 1036, stat: Journal Article,

Apheresis with adacolumn in patients with systemic lupus erythematosus (SLE): A pilot study
Merrill, JT; Buyon, JP; Clancy, R; Petri, M; Becker, M; Komatsu, Y; Gustofson, LM; Wang, T
2004 SEP ;50(9):S416-S416, Arthritis & rheumatism
— id: 49046, year: 2004, vol: 50, page: S416, stat: Journal Article,

Combined oral contraceptives (OC) are not associated with an increased rate of flare in SLE patients in SELENA
Petri, M; Buyon, JP; Kim, M; Kalunian, K; Grossman, J; Hahn, B; Sammaritano, L; Lockshin, M; Merrill, J; Belmont, HM; Askanase, AD; McCune, WJ; Hearth-Holmes, M; Dooley, M; Von Feldt, J; Friedman, A; Tan, M; Davis, J; Cronin, M; Diamond, B; Mackay, M; Sigler, L; Fillius, M
2004 SEP ;50(9):S239-S240, Arthritis & rheumatism
— id: 49033, year: 2004, vol: 50, page: S239, stat: Journal Article,

Effects of prasterone on disease activity and symptoms in women with active systemic lupus erythematosus
Petri, Michelle A; Mease, Philip J; Merrill, Joan T; Lahita, Robert G; Iannini, Mark J; Yocum, David E; Ginzler, Ellen M; Katz, Robert S; Gluck, Oscar S; Genovese, Mark C; Van Vollenhoven, Ronald; Kalunian, Kenneth C; Manzi, Susan; Greenwald, Maria W; Buyon, Jill P; Olsen, Nancy J; Schiff, Michael H; Kavanaugh, Arthur F; Caldwell, Jacques R; Ramsey-Goldman, Rosalind; St Clair, E William; Goldman, Allan L; Egan, Rita M; Polisson, Richard P; Moder, Kevin G; Rothfield, Naomi F; Spencer, Robert T; Hobbs, Kathryn; Fessler, Barri J; Calabrese, Leonard H; Moreland, Larry W; Cohen, Stanley B; Quarles, Betty J; Strand, Vibeke; Gurwith, Marc; Schwartz, Kenneth E
2004 Sep;50(9):2858-2868, Arthritis & rheumatism
OBJECTIVE: To determine whether prasterone administration results in improvement or stabilization of systemic lupus erythematosus (SLE) disease activity and its symptoms. METHODS: Women with active SLE were treated with prasterone 200 mg/day plus standard SLE treatments or with placebo plus standard SLE treatments for up to 12 months in this randomized, double-blind investigation conducted at 27 centers. Standard SLE treatments included prednisone (</=10 mg/day), antimalarials, and immunosuppressive agents; dosages were required to be stable for >/=6 weeks prior to enrollment and remain unchanged during protocol treatment. Responders were patients who experienced no clinical deterioration and had improvement or stabilization over the duration of the study in 2 disease activity measures (the SLE Disease Activity Index [SLEDAI] and the Systemic Lupus Activity Measure) and 2 quality of life measures (patient's global assessment and the Krupp Fatigue Severity Scale). RESULTS: A total of 381 women with SLE were enrolled. Among patients with clinically active disease at baseline (SLEDAI score >2), 86 of 147 in the prasterone group (58.5%) demonstrated improvement or stabilization without clinical deterioration, as compared with 65 of 146 in the placebo group (44.5%) (P = 0.017). Acne and hirsutism were reported in 33% and 16%, respectively, of the prasterone group and in 14% and 2%, respectively, of the placebo group (P < 0.05 for both comparisons). However, most cases of acne and hirsutism were mild and did not require withdrawal from therapy. Myalgias and oral stomatitis were reported less frequently in the prasterone group (22% and 15%, respectively) than in the placebo group (36% and 23%, respectively) (P < 0.05 for both comparisons). Serum levels of high-density lipoprotein cholesterol, triglycerides, and C3 complement significantly decreased, while levels of testosterone and, to a lesser extent, estradiol increased in the prasterone group. CONCLUSION: In adult women with active SLE, administration of prasterone at a dosage of 200 mg/day improved or stabilized signs and symptoms of disease and was generally well tolerated
— id: 73530, year: 2004, vol: 50, page: 2858, stat: Journal Article,

In vivo and in vitro evidence that shedding of endothelial protein c receptors (EPCR) contributes to the pathogenesis of SLE
Sesin, C; Yin, X; Buyon, J; Clancy, R
2004 SEP ;50(9):S647-S648, Arthritis & rheumatism
— id: 49052, year: 2004, vol: 50, page: S647, stat: Journal Article,

Intravenous immunoglobulin and placental transport of anti-Ro/La antibodies: comment on the letter by Kaaja and Julkunen
Tran, Hai B; Cavill, Dana; Buyon, Jill P; Gordon, Tom P
2004 Jan;50(1):337-338, Arthritis & rheumatism
— id: 73531, year: 2004, vol: 50, page: 337, stat: Journal Article,

Development of systemic lupus erythematosus in a patient with congenital heart block - Reply
Askanase, AD; Clancy, RM; Buyon, JP
2003 SEP ;48(9):2699-2699, Arthritis & rheumatism
— id: 55539, year: 2003, vol: 48, page: 2699, stat: Journal Article,

Recurrence rates for neonatal lupus: Data from the US national research registry for neonatal lupus
Askanase, AD; Rupel, A; Solomon, DG; Katholi, M; Buyon, JP
2003 SEP ;48(9):S510-S510, Arthritis & rheumatism
— id: 55444, year: 2003, vol: 48, page: S510, stat: Journal Article,

TGF beta: A fetal factor in the scarring cascade to anti-Ro associated congenital heart block (CHB)
Backer, C; Buyon, J; Clancy, R
2003 SEP ;48(9):S254-S254, Arthritis & rheumatism
— id: 55435, year: 2003, vol: 48, page: S254, stat: Journal Article,

The autoimmune connection : essential information for women on diagnosis, treatment, and getting on with life
Baron-Faust, Rita; Buyon, Jill P
Chicago IL : Contemporary Books, 2003,
— id: 903, year: 2003, vol: , page: , stat: ,

The autoimmune connection : essential information for women on diagnosis, treatment, and getting on with life
Baron-Faust, Rita; Buyon, Jill P
New York : McGraw-Hill, 2003,
— id: 904, year: 2003, vol: , page: , stat: ,

Proposal for a new deffinition of congenital complete atrioventricular block
Brucato, A; Jonzon, A; Friedman, D; Allan, LD; Vignati, G; Gasparini, M; Stein, JI; Montella, S; Michaelsson, M; Buyon, J
2003 JUL ;12(6):427-435, Lupus
The classic old definition of congenital heart block by Yater (1929) is still generally accepted: 'Heart block established in a young patient. There must be some evidence of the existence of the slow pulse at a fairly early age and absence of a history of any infection which might cause the condition after birth: notably diphtheria, rheumatic fever, chorea and congenital syphilis'. However, other definitions are used. We systematically reviewed 1825 cases from 38 separate studies. We conclude that complete AV blocks detected in utero in the absence of structural abnormalities differ from blocks detected later in life with respect to pathogenesis (they are generally associated with maternal anti-Ro/SSA antibodies), poorer childhood prognosis, increased risk of developing late-onset dilated cardiomyopathy, different maternal clinical features and increased risk of recurrence in future pregnancies. For these reasons we propose a new modern definition of congenital complete AV block which might be acceptable to cardiologists, rheumatologists, pediatricians and obstetricians: 'an AV block is defined as congenital if it is diagnosed in utero, at birth or within the neonatal period (0-27 days after birth)'
— id: 37099, year: 2003, vol: 12, page: 427, stat: Journal Article,

From antibody insult to fibrosis in neonatal lupus - the heart of the matter
Buyon, Jill P; Clancy, Robert M
2003 ;5(6):266-270, Arthritis research & therapy
Few diseases exemplify the integration of research from bench to bedside as well as neonatal lupus, often referred to as a model of passively acquired autoimmunity. In essence, this disease encompasses two patients, both the mother and her child. The signature histologic lesion of autoimmune-associated congenital heart block is fibrosis of the conducting tissue, and in some cases the surrounding myocardium. It is astounding how rapid and, in most cases, irreversible is the fibrotic response to injury. The mechanism by which maternal anti-SSA/Ro-SSB/La antibodies initiate and perpetuate inflammation, and eventuate in scarring of the atrioventricular node, is not yet defined. In vitro and in vivo studies suggest that one pathologic cascade leading to scarring may be initiated via apoptosis, resulting in translocation of SSA/Ro-SSB/La antigens and subsequent surface binding by maternal autoantibodies. These opsonized cardiocytes are phagocytosed by macrophages, which secrete factors that transdifferentiate fibroblasts into myofibroblasts, a scarring phenotype. Dissecting the individual components in this fibrotic pathway should provide insights into the rarity of irreversible injury and should form the basis of rational approaches to prevention and treatment
— id: 46212, year: 2003, vol: 5, page: 266, stat: Journal Article,

Maternal autoantibodies and congenital heart block: mediators, markers, and therapeutic approach
Buyon, Jill P; Clancy, Robert M
2003 May 2;33(3):140-154, Seminars in arthritis & rheumatism
— id: 46222, year: 2003, vol: 33, page: 140, stat: Journal Article,

Neonatal lupus syndromes
Buyon, Jill P; Clancy, Robert M
2003 Sep;15(5):535-541, Current opinion in rheumatology
The neonatal lupus syndromes, although quite rare, provide an excellent opportunity to examine disease from bench to bedside. During the past year numerous publications have reported basic and clinical research. Although anti-SSA/Ro-SSB/La antibodies are detected in more than 85% of mothers whose fetuses are identified with conduction abnormalities in a structurally normal heart, when clinicians applied this testing to their pregnant patients, the risk for a woman with the candidate antibodies to have a child with congenital heart block was at or below one in 50. Although the precise pathogenic mechanism of antibody-mediated injury remains unknown, it is clear that the antibodies alone are insufficient to cause disease, and fetal factors are likely contributory. In vivo and in vitro evidence supports a pathologic cascade involving apoptosis of cardiocytes, surface translocation of Ro and La antigens, binding of maternal autoantibodies, secretion of profibrosing factors from the scavenging macrophages, and transdifferentiation of cardiac fibroblasts to a myofibroblast scarring phenotype. Cross-reactivity of anti-52-kD SSA/Ro antibodies with a serotoninergic cardiac receptor, 5-hydroxytryptamine (HT)4, has been suggested but remains unconfirmed. The spectrum of cardiac abnormalities continues to grow, with varying degrees of block identified in utero and reports of late-onset cardiomyopathy (some of which display endocardial fibroelastosis). Moreover, there is now clear documentation that incomplete blocks (including those improving in utero with dexamethasone) can progress postnatally, despite the clearance of the maternal antibodies from the neonatal circulation. Better echocardiographic measurements that identify first-degree block in utero may be the optimal means of approaching pregnant women at risk. Prophylactic therapies, including treatment with intravenous immunoglobulin, await larger trials. Reassuringly, most children with neonatal lupus syndromes do not develop rheumatic diseases, although follow-up is limited to late adolescence. To further efforts both at the bench and bedside, national research registries established in the United States and Canada are critical
— id: 39085, year: 2003, vol: 15, page: 535, stat: Journal Article,

Neonatal lupus: review of proposed pathogenesis and clinical data from the US-based Research Registry for Neonatal Lupus
Buyon, Jill P; Clancy, Robert M
2003 Feb;36(1):41-50, Autoimmunity
Congenital heart block (CHB), a life-threatening manifestation of neonatal lupus, offers aunique opportunity to study the effect or arm of immunity and define the pathogenicity of an autoantibody in mediating tissue injury. This review focuses on our recent in vitro model which supports a cascade from antibody insult to unchecked fibrosis. In brief, it is proposed that the fetal cardiac myocyte undergoes apoptosis which facilitates transfer of intracellular Ro and La antigens to the surface where they are bound by circulating maternal autoantibodies (anti-SSA/Ro-SSB/La antibodies). Scavenging macrophages phagocytose these inadvertently 'opsonized' cardiocytes, leading to the secretion of pro-inflammatory and pro-fibrotic cytokines, the latter of which transdifferentiate fibroblasts into myofibroblasts and thereby promote scarring. Immunohistologic study of a heart from a neonate dying of CHB supports this model in that macrophages and myofibroblasts were demonstrated. To facilitate both basic and clinical research, a Research Registry for Neonatal Lupus was established in 1994 by the U.S. National Institute of Arthritis, Musculoskeletal and Skin Diseases. Maternal and fetal outcomes are addressed as well as recurrence rates. Laboratory evaluation and management decisions during pregnancy are provided
— id: 39218, year: 2003, vol: 36, page: 41, stat: Journal Article,

Autoantibody-associated congenital heart block: the clinical perspective
Buyon, Jill P; Friedman, Deborah M
2003 Nov;5(5):374-378, Current rheumatology reports
Congenital heart block (CHB) can occur in association with structural heart disease, such as atrioventricular septal defects, left atrial isomerism, and abnormalities of the great arteries, with tumors, such as mesotheliomas, or as an isolated defect. In 1928, Aylward reported the occurrence of CHB in two children whose mother 'suffered from Mikulicz's disease.' This curious clinical observation was further solidified by the 1970s, with reports of CHB in children whose mothers had autoimmune diseases and that the maternal sera contained antibodies to Ro ribonucleoproteins. It was subsequently reported that many mothers also had antibodies to La. Other abnormalities affecting the skin, liver, and blood elements were associated with anti-Ro/La antibodies in the maternal and fetal circulation, and are now grouped under the heading of neonatal lupus syndromes. Neonatal lupus was termed because the cutaneous lesions of the neonate resembled those seen in systemic lupus erythematosus
— id: 46288, year: 2003, vol: 5, page: 374, stat: Journal Article,

Congenital heart block: do fetal factors fuel the fire from inflammation to fibrosis?
Buyon, Jill P; Rupel, Ann; Clancy, Robert M
2003 ;12(10):731-734, Lupus
— id: 73534, year: 2003, vol: 12, page: 731, stat: Journal Article,

Prevention of recurrence of congenital heart block with intravenous immunoglobulin and corticosteroid therapy: comment on the editorial by Buyon et al - Reply
Buyon, JP; Kim, MY; Copel, JA; Friedman, DA
2003 JAN ;48(1):281-282, Arthritis & rheumatism
— id: 55590, year: 2003, vol: 48, page: 281, stat: Journal Article,

Fetal factors in neonatal lupus (NL): Association of a promoter polymorphism of tumor necrosis factor-alpha and HLA-DRB1 haplotype in children with rash
Clancy, R; Backer, C; Cohen, S; Yin, X; Buyon, J
2003 SEP ;48(9):S436-S436, Arthritis & rheumatism
— id: 55441, year: 2003, vol: 48, page: S436, stat: Journal Article,

Expression of endothelial cell protein C receptor and inducible nitric oxide synthase (iNOS) by circulating endothelial cells (CEC) in Systemic Lupus Erythematosus (SLE)
Clancy, R; Sesin, C; Kurosawa, S; Buyon, J
2003 SEP ;48(9):S374-S374, Arthritis & rheumatism
— id: 55439, year: 2003, vol: 48, page: S374, stat: Journal Article,

HLA-DRB1 relationships in neonatal lupus (NL) families
Clancy, R; Yin, X; Askanase, A; Miranda-Carus, E; Nelson, JL; Sestak, A; Harley, J; Buyon, J
2003 SEP ;48(9):S410-S411, Arthritis & rheumatism
— id: 55440, year: 2003, vol: 48, page: S410, stat: Journal Article,

Cytokine polymorphisms and histologic expression in autopsy studies: contribution of TNF-alpha and TGF-beta1 to the pathogenesis of autoimmune-associated congenital heart block
Clancy, Robert M; Backer, Chelsea B; Yin, Xiaoming; Kapur, Raj P; Molad, Yair; Buyon, Jill P
2003 Sep 15;171(6):3253-3261, Journal of immunology
Although Abs to SSA/Ro-SSB/La are necessary for the development of congenital heart block (CHB), the low frequency suggests that fetal factors are contributory. Because CHB involves a cascade from inflammation to scarring, polymorphisms of the TNF-alpha promoter region and codons 10 and 25 of the TGF-beta gene were evaluated in 88 children (40 CHB, 17 rash, 31 unaffected siblings) and 74 mothers from the Research Registry for Neonatal Lupus (NL). Cytokine expression was assessed in autopsy material from two fetuses with CHB. Significantly increased frequency of the -308A (high-producer) allele of TNF-alpha was observed in all NL groups compared with controls. In contrast, the TGF-beta polymorphism Leu(10) (associated with increased fibrosis) was significantly higher in CHB children (genotypic frequency 60%, allelic frequency 78%) than unaffected offspring (genotypic frequency 29%, p = 0.016; allelic frequency 56%, p = 0.011) and controls, while there were no significant differences between controls and other NL groups. For the TGF-beta polymorphism, Arg(25), there were no significant differences between NL groups and controls. In fetal CHB hearts, protein expression of TGF-beta, but not TNF-alpha, was demonstrated in septal regions, extracellularly in the fibrous matrix, and intracellularly in macrophage infiltrates. Age-matched fetal hearts from voluntary terminations expressed neither cytokine. TNF-alpha may be one of several factors that amplify susceptibility; however, the genetic studies, backed by the histological data, more convincingly link TGF-beta to the pathogenesis of CHB. This profibrosing cytokine and its secretion/activation circuitry may provide a novel direction for evaluating fetal factors in the development of a robust animal model of CHB as well as therapeutic strategies in humans
— id: 39087, year: 2003, vol: 171, page: 3253, stat: Journal Article,

Clearance of apoptotic cells: TGF-beta in the balance between inflammation and fibrosis
Clancy, Robert M; Buyon, Jill P
2003 Dec;74(6):959-960, Journal of leukocyte biology
Transforming growth factor-beta (TGF-beta) has been considered an anti-inflammatory cytokine responsible for the bland removal of apoptotic cells. What is less established is the extent of secretion of this cytokine during the clearance of opsonized apoptotic cells via Fcgamma-mediated uptake. To date both decreased (favoring predominance of inflammation) and increased (favoring resolution of inflammation but potentially pro-fibrotic) responses have been demonstrated. IN an in vitro model of autoantibody-induced cardiac injury, we herein demonstrate that macrophages cocultured with apoptotic human fetal cardiocytes bound by anti-SSA/Ro antibodies secrete increased levels of TGF-beta. Prolonged secretion of this cytokine may contribute to the exuberant scarring seen in congenital heart block associated with maternal autoantibodies reactive with SSA/Rho and SSB/La antigens
— id: 48186, year: 2003, vol: 74, page: 959, stat: Journal Article,

Does the cellular localization of antigens in or on apoptotic blebs influence the pathogenicity or benefit of cognate antibodies? Comment on the article by Dieude et al
Clancy, Robert M; Chan, Edward K L; Chandrashekhar, Sarayu; Buyon, Jill P
2003 Jul;48(7):2080-2082, Arthritis & rheumatism
— id: 73536, year: 2003, vol: 48, page: 2080, stat: Journal Article,

Fetal cardiac function assessed by Doppler myocardial performance index (Tei Index)
Friedman, D; Buyon, J; Kim, M; Glickstein, J S
2003 Jan;21(1):33-36, Ultrasound in obstetrics & gynecology
OBJECTIVES: The Tei Index (TI) is a useful, non-invasive, Doppler-derived myocardial performance tool which can be used to assess aspects of systolic and diastolic function. The aim of this study was to determine normal values of fetal left ventricular (LV) TI in second- and third- trimester fetuses and to compare these to other values reported in the literature. METHODS: Doppler waveforms of the LV outflow tracts were obtained in 74 second- and early third-trimester fetuses. The LV isovolumic contraction time (ICT), isovolumic relaxation time (IRT) and ejection time (ET) were measured and the TI calculated using the formula (ICT + IRT)/ET. The literature on LV myocardial function in the fetus was also reviewed. RESULTS: The normal TI in second- and early third-trimester fetuses (18-31 weeks' gestation) was 0.53 +/- 0.13. The ICT was 43 +/- 14 ms, the ET was 173 +/- 16 ms and the IRT was 48 +/- 13 ms. CONCLUSION: The TI can be easily obtained in the fetus without the need for precise anatomic imaging. The TI may be a useful tool to explore fetal myocardial function in different clinical situations
— id: 132526, year: 2003, vol: 21, page: 33, stat: Journal Article,

Birth order, gender and recurrence rate in autoantibody-associated congenital heart block: implications for pathogenesis and family counseling
Solomon, Daniel G; Rupel, Ann; Buyon, Jill P
2003 ;12(8):646-647, Lupus
— id: 73535, year: 2003, vol: 12, page: 646, stat: Journal Article,

Myocardial-tissue-specific phenotype of maternal microchimerism in neonatal lupus congenital heart block
Stevens, Anne M; Hermes, Heidi M; Rutledge, Joe C; Buyon, Jill P; Nelson, J Lee
2003 Nov 15;362(9396):1617-1623, Lancet
BACKGROUND: During pregnancy, maternal cells pass into the fetus, where they can persist for many years after birth. We investigated the presence of maternal cells in neonatal lupus syndrome (NLS), an autoimmune disease that develops in utero. The most serious complication of NLS is inflammation of the atrioventricular node leading to congenital heart block (CHB). METHODS: In a blinded case-control study, maternal (female) cells were detected and quantified in male NLS and control heart-tissue samples. We used fluorescence in-situ hybridisation to label X and Y chromosomes. Studies in transplantation suggest that donor cells can differentiate into somatic tissue cells. Therefore, we asked whether maternal cells transferred in utero have cellular plasticity. To simultaneously identify and characterise maternal cells, we developed a technique by which multiple phenotypic markers could be detected concurrently with fluorescence in-situ hybridisation in the same cells of a tissue section. FINDINGS: Maternal cells were found in 15 of 15 sections of NLS heart tissue, ranging from 0.025% to 2.2% of total cells. By contrast, maternal cells were found in two of eight control sections (0-0.1%). Very few maternal cells expressed the haemopoietic cell marker CD45. Most expressed sarcomeric alpha actin, a specific marker for cardiac myocytes. INTERPRETATION: Our findings suggest that differentiated tissue-specific maternal microchimerism can occur in neonates. Thus, semiallogeneic maternal cells could be the target of an immune response. Alternatively, maternal cells could contribute to a secondary process of tissue repair
— id: 73533, year: 2003, vol: 362, page: 1617, stat: Journal Article,

Moderate dose steroids prevent severe flares in a prospective Multicenter study of serologically active, clinically stable systemic lupus erythematosus (SLE) patients
Tseng, C; Buyon, J; Kim, M; Belmont, HM; Mackay, M; Diamond, B; Marder, G; Rosenthal, P; Haines, K; Abramson, S
2003 SEP ;48(9):S260-S261, Arthritis & rheumatism
— id: 55436, year: 2003, vol: 48, page: S260, stat: Journal Article,

Spectrum and progression of conduction abnormalities in infants born to mothers with anti-SSA/Ro-SSB/La antibodies
Askanase, A D; Friedman, D M; Copel, J; Dische, M R; Dubin, A; Starc, T J; Katholi, M C; Buyon, J P
2002 ;11(3):145-151, Lupus
The classic cardiac manifestation of neonatal lupus is congenital heart block, attributed to antibody-mediated inflammation and subsequent fibrosis of the atrioventricular (AV) node. In considering the pathologic process of injury it may be that tissue damage results in a range of conduction abnormalities. Identification of less-advanced degrees of block or of fibrosis around the AV node without any conduction abnormality on EKG would support this pathologic model, and serve as a potential marker for treatment if the conduction defect could be shown to progress. To ascertain the spectrum of arrhythmias associated with maternal anti-SSA/Ro-SSB/La antibodies, records of all children enrolled in the Research Registry for Neonatal Lupus were reviewed. Of 187 children with congenital heart block whose mothers have anti-SSA/Ro-SSB/La antibodies, nine had a prolonged PR interval on EKG at birth, four of whom progressed to more advanced AV block. A child whose younger sibling had third degree block was diagnosed with first degree block at age 10 years at the time of surgery for a broken wrist. Two children diagnosed in utero with second degree block were treated with dexamethasone and reverted to normal sinus rhythm by birth, but ultimately progressed to third degree block. Four children had second degree block at birth: of these, two progressed to third degree block. Sinus bradycardia (< 100 bpm) was present in three (3.8%) of 78 fetuses for whom atrial rates were recorded by echocardiogram. Of 40 neonates for whom EKGs were available, the mean atrial rate was 137+/-20 bpm (range 75-200). These data have important research and clinical implications. In contrast to the AV node, permanent sinoatrial nodal involvement is not clinically apparent. Perhaps many fetuses sustain mild inflammation, but resolution is variable, as suggested by the presence of incomplete AV block. Since subsequent progression of less-advanced degrees of block can occur, an EKG should be performed on all infants born to mothers with anti-SSA/Ro-SSB/La antibodies
— id: 39649, year: 2002, vol: 11, page: 145, stat: Journal Article,

Reproductive health in SLE
Askanase, Anca D; Buyon, Jill P
2002 Apr;16(2):265-280, Bailliere's best practice & research. Clinical rheumatology
Oral contraceptives containing oestrogens and hormone replacement therapy are generally not prescribed for women with systemic lupus erythematosus (SLE). The concern regarding oestrogens is based on the greater incidence of SLE in women, abnormalities of oestrogen metabolism, murine models of lupus, several anecdotes of patients having disease flares while receiving hormones, and one retrospective study in patients with pre-existing renal disease. For healthy women and those with SLE, there are clinical settings in which exogenous oestrogens provide benefit. For pre-menopausal women, these include provision of safe and effective birth control, protection against bone loss, and the consideration of oral contraceptives to preserve fertility in patients taking cyclophosphamide. For post-menopausal women, these include treatment of hot flushes and vaginal dryness, prevention of osteoporosis and, more controversial, prevention of atherosclerosis. Other exogenous hormones (clomiphene citrate, gonadotropins, gonadotropin-releasing hormones) may be used to elevate levels of endogenous oestrogen and stimulate ovulation in patients with diminished fertility.This chapter focuses on three broad categories: birth control, assisted reproduction and hormone replacement
— id: 39637, year: 2002, vol: 16, page: 265, stat: Journal Article,

The presence of IgG antibodies reactive with components of the SSA/Ro-SSB/La complex in human breast milk: implications in neonatal lupus
Askanase, Anca D; Miranda-Carus, M Eugenia; Tang, Xiaoyin; Katholi, Margaret; Buyon, Jill P
2002 Jan;46(1):269-271, Arthritis & rheumatism
— id: 32695, year: 2002, vol: 46, page: 269, stat: Journal Article,

Increased frequency of the high producer genotype TT (Codon 10) of TGF beta in congenital heart block and TNF2 allele in families with neonatal lupus
Backer, C; Loma-Sanner, I; Yin, X; Chandrashekhar, S; Sullivan, K; Clancy, R; Buyon, J
2002 SEP ;46(9):S621-S621, Arthritis & rheumatism
— id: 55555, year: 2002, vol: 46, page: S621, stat: Journal Article,

Role of NFkB and Rac in fibroblast activation: Key signaling molecules in the pathogenesis of congenital heart block (CAB)
Burg, N; Ross, C; Askanase, A; Buyon, J; Clancy, R
2002 SEP ;46(9):S254-S255, Arthritis & rheumatism
— id: 55553, year: 2002, vol: 46, page: S254, stat: Journal Article,

Cardiac 5-HT(4) Serotoninergic Receptors, 52kD SSA/Ro and Autoimmune-Associated Congenital Heart Block
Buyon, Jill P; Clancy, Robert; Di Donato, Francis; Miranda-Carus, M Eugenia; Askanase, Anca D; Garcia, Joanne; Qu, Yongxia; Hu, Keli; Yue, Yuankun; Chan, Edward K L; Boutjdir, Mohamed
2002 Aug;19(1-2):79-79, Journal of autoimmunity
It was recently reported that sera from patients with systemic lupus erythematosus contain antibodies reactive with the second extracellular loop of the serotoninergic 5-HT(4) receptor expressed in the human heart. This antibody response was associated with antibodies to 52kD SSA/Ro, a reactivity prevalent in mothers of children with congenital heart block (CHB). The current study was undertaken to determine whether the 5-HT(4) receptor is a target of the immune response in these mothers. Initial experiments demonstrated mRNA expression of the 5-HT(4) receptor in the human foetal atrium. Electrophysiologic studies established that human foetal atrial cells express functional 5-HT(4) receptors. Sera from 116 mothers enrolled in the Research Registry for Neonatal Lupus, whose children have CHB, were evaluated. Ninety-nine (85%) of these maternal sera contained antibodies to SSA/Ro, 84% of which were reactive with the 52kD SSA/Ro component by immunoblot. None of the 116 sera were reactive with the peptide spanning aa165-185 of the serotoninergic receptor. Rabbit antisera which recognized this peptide did not react with 52kD SSA/Ro or peptide aa365-382 in the C terminus. Although 5-HT(4) receptors are present and functional in the human foetal heart, maternal antibodies to the 5-HT(4) receptor are not associated with the development of CHB
— id: 32693, year: 2002, vol: 19, page: 79, stat: Journal Article,

Transdifferentiation of cardiac fibroblasts, a fetal factor in anti-SSA/Ro-SSB/La antibody-mediated congenital heart block
Clancy, Robert M; Askanase, Anca D; Kapur, Raj P; Chiopelas, Efstathia; Azar, Natalie; Miranda-Carus, M Eugenia; Buyon, Jill P
2002 Aug 15;169(4):2156-2163, Journal of immunology
The signature lesion of autoantibody-associated congenital heart block (CHB) is fibrosis of the conducting tissue. To date, participation of myofibroblasts in the cascade to injury has been unexplored. The importance of myofibroblast/macrophage cross-talk is demonstrated by the novel finding of these cell types in the heart of a neonate dying of CHB. This clue to pathogenesis prompted consideration of the mechanism by which maternal anti-SSA/Ro-SSB/La Abs initiate an inflammatory response and promote fibrosis. Isolated cardiocytes from 16-24 wk abortuses were rendered apoptotic by exposure to poly (2-) hydroxyethylmethacrylate; flow cytometry confirmed surface expression of Ro/La. Apoptotic cardiocytes were incubated with affinity-purified Abs to 52 and 60 kDa Ro from CHB mothers (opsonized) or IgG fractions from healthy donors (nonopsonized). Macrophages cultured with opsonized apoptotic cardiocytes expressed proinflammatory markers, supported by a three-fold increase in active alpha(V)beta(3) integrin. Fetal cardiac fibroblasts exposed to supernatants obtained from macrophages incubated with opsonized apoptotic cardiocytes (but not nonopsonized) dramatically increased expression of the myofibroblast marker alpha-smooth muscle actin (SMAc). The 'opsonized' supernatant reversed an inhibitory effect of the 'nonopsonized' supernatant on proliferation of fibroblasts (120 vs 69%, p < 0.05). Parallel experiments examined the effects of two cytokines and their neutralizing Abs on fibroblasts. TGFbeta1 increased SMAc staining but decreased proliferation. TNF-alpha did not affect either readout. Addition of anti-TGFbeta1 Abs to the 'opsonized' supernatant blocked SMAc expression but increased proliferation, while anti-TNF-alpha blocking Abs had no effects. These data suggest that transdifferentiation of cardiac fibroblasts to a scarring phenotype is a pathologic process initiated by maternal Abs
— id: 32694, year: 2002, vol: 169, page: 2156, stat: Journal Article,

Congenital heart block in neonatal lupus: the pediatric cardiologist's perspective
Friedman, Deborah M; Rupel, Ann; Glickstein, Julie; Buyon, Jill P
2002 Jun;69(6):517-522, Indian journal of pediatrics
CLINICAL PRESENTATION: Congenital heart block (CHB) in the absence of major structural abnormalities is associated with maternal antibodies to Ro (SS-A) and La (SS-B). CHB is most commonly diagnosed between 18 and 24 wk of gestation, and may be first, second or third degree (complete). Mortality approaches approximately 20%, and most surviving children require pacemakers. Affected infants may develop cardiomyopathy. Abnormalities in the skin, liver and blood of neonates are also associated with anti-Ro/La antibodies, and are usually self-limiting; these manifestations and CHB are collectively referred to as neonatal lupus syndromes (NLS). INVESTIGATION OF PATHOGENESIS: Recent studies demonstrate that Ro/La ribonucleoproteins appear on the surface of apoptotic fetal cardiocytes and are recognized by their cognate antibodies, promoting an inflammatory response. Mice immunized with Ro/La proteins have offspring with conduction abnormalities. In vitro, human serum and IgG with anti-Ro/La antibodies affect the conducting properties of isolated animal heart tissue. DIAGNOSTIC PROBLEMS: If fetal bradycardia is identified, a 2-dimensional and M-mode fetal echocardiographic and Doppler ultrasound should be obtained to determine whether there is an atrial arrhythmia or atrioventricular (AV) block, and to what degree, and whether there are major structural abnormalities of the heart. The mother's serum should be tested by ELISA for anti-Ro and/or anti-La antibodies. THERAPEUTIC OPTIONS: To date, only anecdotal and retrospective evidence guides in utero therapy of CHB. A prospective trial is currently underway to evaluate the efficacy of maternal oral dexamethasone in treating newly identified first, second or third degree block. Established third-degree block appears to be irreversible. Dexamethasone and sympathomimetics may be of some benefit in treating hydrops fetalis. In pregnant women with anti-Ro/La antibodies, prophylactic therapy is not indicated but serial echocardiographic analysis is strongly recommended, with emphasis on the mechanical PR interval to identify a reversible block. CONCLUSION: CHB occurs in approximately 1-5% of pregnancies in mothers with anti-Ro/La antibodies, independent of the mother's disease status, and in approximately 15-20% of pregnancies following the birth of a child with NLS. Treatment of CHB identified in utero is not established but guidelines are provided. Serial echocardiographic monitoring of high-risk pregnancies, using the mechanical PR interval to identify first degree block, may afford the earliest opportunities for therapeutic intervention
— id: 73538, year: 2002, vol: 69, page: 517, stat: Journal Article,

Circulating endothelial cells (CEC) in systemic lupus erythematosus: Activation markers and relationship to disease activity
Lee, E; Sesin, C; Abramson, S; Buyon, J; Clancy, R
2002 SEP ;46(9):S214-S214, Arthritis & rheumatism
— id: 55551, year: 2002, vol: 46, page: S214, stat: Journal Article,

Hepatobiliary disease in neonatal lupus: prevalence and clinical characteristics in cases enrolled in a national registry
Lee, Lela A; Sokol, Ronald J; Buyon, Jill P
2002 Jan;109(1):E11-E11, Pediatrics
OBJECTIVE: To extend the information base on the hepatobiliary manifestations of neonatal lupus erythematosus (NLE) with regard to frequency of occurrence, clinical characteristics, and outcome. METHODS: Review of records from the Research Registry for Neonatal Lupus. RESULTS: Nineteen (9%) of 219 patients who had NLE and were enrolled in a national registry had probable or possible NLE hepatobiliary disease. In 16 cases, hepatobiliary disease occurred in addition to cardiac or cutaneous NLE. In 3 cases, hepatobiliary disease occurred as the sole clinical manifestation of NLE. Three clinical variants of hepatobiliary disease were observed: 1) severe liver failure present during gestation or in the neonatal period, often with the phenotype of neonatal iron storage disease; 2) conjugated hyperbilirubinemia with mild or no elevations of aminotransferases, occurring in the first few weeks of life; and 3) mild elevations of aminotransferases occurring at approximately 2 to 3 months of life. The prognosis for the children in the last 2 categories is excellent. CONCLUSIONS: Hepatobiliary disease is a relatively common finding in NLE and can be the sole clinical manifestation of NLE. Clinicians should be aware of the broad range of hepatobiliary disease that may occur in children with NLE
— id: 73542, year: 2002, vol: 109, page: E11, stat: Journal Article,

The rate and pattern of organ damage in late onset systemic lupus erythematosus
Maddison, Peter; Farewell, Vernon; Isenberg, David; Aranow, Cynthia; Bae, Sang-Cheol; Barr, Susan; Buyon, Jill; Fortin, Paul; Ginzler, Ellen; Gladman, Dafna; Hanly, John; Manzi, Susan; Nived, Ola; Petri, Michelle; Ramsey-Goldman, Rosalind; Sturfelt, Gunnar
2002 May;29(5):913-917, Journal of rheumatology
OBJECTIVE: To compare the extent and type of damage in patients with late onset and earlier onset systemic lupus erythematosus (SLE) using the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SDI). METHODS: A total of 86 SLE patients with disease onset after the age of 54 years were matched for center, sex, and ethnic origin with 155 SLE patients with disease onset before the age of 40 years. SDI scores were obtained at one year and 5 years after the diagnosis of SLE. Analysis was based on conditional logistic regression. RESULTS: SDI scores were higher in the late onset group than in younger patients at both one [mean 0.7 (range 0-3) vs 0.3 (range 0-3); p < 0.001] and 5 years [mean 1.6 (range 0-8) vs 0.9 (range 0-7); p < 0.001] after diagnosis. There was also a difference in the pattern of organ damage. While damage to the skin, kidneys, and central nervous system occurred with similar frequency, late onset disease was characterized by significantly more cardiovascular (OR 14.13, p < 0.001), ocular (OR 9.38, p = 0.001), and musculoskeletal (OR 2.68, p = 0.016) damage and malignancy (OR 7.04, p = 0.046). CONCLUSION: The occurrence of organ damage assessed by the SDI is greater in patients with late onset SLE than in younger patients and, by this criterion, lupus cannot be judged to be more benign in this age group. Also, the pattern of damage is different, but whether this reflects age per se or the effect of the disease in the elderly remains to be established
— id: 73540, year: 2002, vol: 29, page: 913, stat: Journal Article,

Long-term followup of children with neonatal lupus and their unaffected siblings
Martin, Victor; Lee, Lela A; Askanase, Anca D; Katholi, Margaret; Buyon, Jill P
2002 Sep;46(9):2377-2383, Arthritis & rheumatism
OBJECTIVE: To determine in a longitudinal cohort study whether children with varied manifestations of neonatal lupus or their unaffected siblings later develop autoantibodies and/or rheumatic diseases. METHODS: To obtain information on the health of children ages >or=8 years who had manifestations of neonatal lupus (affected group) and their unaffected siblings (unaffected group), questionnaires were sent to mothers (with anti-SSA/Ro and/or anti-SSB/La antibodies) who were enrolled in the National Institute of Arthritis and Musculoskeletal and Skin Diseases/Hospital for Joint Diseases Research Registry for Neonatal Lupus. Children of healthy mothers referred by the Registry enrollees comprised the control group. Further data were provided by review of medical records. RESULTS: Fifty-five mothers enrolled in the Registry returned questionnaires on 49 children with neonatal lupus and their 45 unaffected siblings. Six children with definite rheumatic/autoimmune diseases were identified: 2 with juvenile rheumatoid arthritis, 1 with Hashimoto thyroiditis, 1 with psoriasis and iritis, 1 with diabetes mellitus and psoriasis, and 1 with congenital hypothyroidism and nephrotic syndrome. All had neonatal lupus, and their mothers had manifestations of autoimmune diseases (Sjogren's syndrome in 4, systemic lupus erythematosus/Sjogren's syndrome in 1, and undifferentiated autoimmune disease in 1). Antinuclear antibodies were present in 4 of 55 sera tested (2 of 33 affected children and 2 of 22 unaffected children). No serum contained antibodies reactive with SSA/Ro or SSB/La antigens. CONCLUSION: These data suggest that children with neonatal lupus require continued followup, especially prior to adolescence and if the mother herself has an autoimmune disease. While there was no apparent increased risk of systemic lupus erythematosus, the development of some form of autoimmune disease (systemic or organ-specific) in early childhood may be of concern. During adolescence and young adulthood, individuals with neonatal lupus and their unaffected siblings do not appear to have an increased risk of developing systemic rheumatic diseases
— id: 39584, year: 2002, vol: 46, page: 2377, stat: Journal Article,

Validation of the Doppler PR interval in the fetus
Rosenthal, David; Friedman, Deborah M; Buyon, Jill; Dubin, Anne
2002 Sep;15(9):1029-1030, Journal of the American Society of Echocardiography
— id: 73537, year: 2002, vol: 15, page: 1029, stat: Journal Article,

Anti-La/SSB antibodies transported across the placenta bind apoptotic cells in fetal organs targeted in neonatal lupus
Tran, Hai B; Macardle, Peter J; Hiscock, Jenny; Cavill, Dana; Bradley, John; Buyon, Jill P; Gordon, Tom P
2002 Jun;46(6):1572-1579, Arthritis & rheumatism
OBJECTIVE: To determine whether La and/or Ro epitopes on apoptotic cells in fetal organs that are targeted in neonatal lupus syndrome (NLS) are accessible for binding by autoantibodies in vivo, we traced the fate of transplacental autoantibodies in a murine passive transfer model. METHODS: Pregnant mice at day 15 of gestation (E15) were injected intraperitoneally with human anti-Ro/La-positive sera or control sera, and transplacental transfer of human autoantibodies was tested by enzyme-linked immunosorbent assay with recombinant antigens. Multiple cryostat sections at the level of the heart of E17 fetuses were visualized simultaneously for human IgG binding and apoptosis (TUNEL) under confocal microscopy. Serial paraffin sections of E17 and E19 fetuses were examined for histologic evidence of inflammation. RESULTS: Human IgG anti-52-kd Ro, anti-60-kd Ro, and anti-La autoantibodies were transported efficiently into the fetal circulation. Human IgG-apoptotic cell complexes were detected in the heart (atrial trabeculae and atrioventricular node), skin, liver, and newly forming bone of fetuses from mothers injected with anti-Ro/La sera but not control sera. The IgG binding was fetal-specific and organ-specific; transplacental autoantibodies did not bind to apoptotic cells in the fetal thymus, lung, brain, or gut. The complexes were not associated with an inflammatory reaction. Injection of mothers with affinity-purified anti-La autoantibodies (but not anti-Ro/La Ig depleted of anti-La) revealed an identical location of IgG binding to apoptotic cells in the fetuses. CONCLUSION: This is the first study to demonstrate that transplacental anti-La autoantibodies bind specifically to apoptotic cells in selected fetal organs in vivo, similar to the organ involvement in NLS. We hypothesize that additional factors are required to promote proinflammatory clearance of IgG-apoptotic cell complexes and subsequent tissue damage
— id: 73539, year: 2002, vol: 46, page: 1572, stat: Journal Article,

Subcellular redistribution of la/SSB autoantigen during physiologic apoptosis in the fetal mouse heart and conduction system: a clue to the pathogenesis of congenital heart block
Tran, Hai B; Ohlsson, Maria; Beroukas, Dimitra; Hiscock, Jenny; Bradley, John; Buyon, Jill P; Gordon, Tom P
2002 Jan;46(1):202-208, Arthritis & rheumatism
OBJECTIVE: In isolated congenital heart block, the mechanism by which maternal autoantibodies target the intracellular components of the Ro/La RNP complex is unclear. Previous studies have demonstrated that cultured fetal cardiac myocytes rendered apoptotic bind antibodies to 48-kd La/SSB. This study further investigated the subcellular distribution of the La antigen during apoptosis in the fetal mouse heart and conduction system. METHODS: The atrioventricular (AV) node, AV bundle, and sinoatrial (SA) node were identified in serial sections prepared from paraffin blocks of normal mouse fetuses on days 15, 17, and 19 of gestation. Apoptosis was detected by TUNEL assay. Under confocal microscopy, fluorescent labeling of fragmented DNA in apoptotic cells was assessed by TUNEL, and La protein localization was visualized simultaneously using a murine monoclonal antibody or affinity-purified human polyclonal anti-La antibodies. RESULTS: Apoptotic cells were detected in and at the periphery of the AV and SA nodes as well as in the fetal heart valve insertions and working myocardium. In contrast, no apoptosis was detected in the adult heart AV node or surrounding myocardium. As expected, the La antigen was predominantly immunolocalized to the nucleus in nonapoptotic cells. However, apoptotic cells showed a marked reduction of nuclear La and redistribution of La to the cytoplasm. High-resolution confocal microscopy revealed that in cells that had undergone apoptosis, La antigen asymmetrically clustered near the surface of TUNEL-positive nuclei and apoptotic bodies. CONCLUSION: These data provide the first in vivo demonstration of the subcellular translocation of La autoantigen during apoptosis in the fetal heart and the conduction system under physiologic conditions. This observation supports the hypothesis that subcellular redistribution of La in the normally developing heart facilitates the binding of cognate maternal antibodies and subsequent tissue damage
— id: 73541, year: 2002, vol: 46, page: 202, stat: Journal Article,

C3, C4, CH50, anti-dsDNA antibodies and C3a complement split products are specific but not sensitive for predicting flares of systemic lupus (SLE)
Tseng, CE; Abramson, SB; Kim, M; Blemont, HM; Haines, K; Petri, M; Buyon, JP
2002 SEP ;46(9):S214-S214, Arthritis & rheumatism
— id: 37119, year: 2002, vol: 46, page: S214, stat: Journal Article,

Cardiac 5-HT4 serotoninergic receptors, 52kD SSA/Ro and autoimmune-associated congenital heart block
Boutjdir, M; Qu, YX; Hu, KL; Di Donato, F; Miranda-Carus, E; Askanase, AD; Garcia, J; Chan, EK; Buyon, JP
2001 OCT 23 ;104(17):752-752, Circulation
— id: 54805, year: 2001, vol: 104, page: 752, stat: Journal Article,

Anti-Ro/SSA antibodies and congenital heart block: necessary but not sufficient
Buyon, J P; Kim, M Y; Copel, J A; Friedman, D M
2001 Aug;44(8):1723-1727, Arthritis & rheumatism
— id: 73545, year: 2001, vol: 44, page: 1723, stat: Journal Article,

Circulating activated endothelial cells in systemic lupus erythematosus: further evidence for diffuse vasculopathy
Clancy R; Marder G; Martin V; Belmont HM; Abramson SB; Buyon J
2001 May;44(5):1203-1208, Arthritis & rheumatism
OBJECTIVE: In flares of systemic lupus erythematosus (SLE), endothelial cells (EC; activated by immune stimuli) are potential participants in the inflammatory processes that contribute to tissue damage. Accordingly, elevated levels of circulating endothelial cells (CEC) may be a marker for vascular injury. This study was undertaken to examine the possibility that stimulated EC are found in the circulation in patients with active SLE. METHODS: The study cohort included 38 patients with SLE and 16 healthy controls. Immunostaining was performed on mononuclear isolates, using mouse P1H12 (endothelial-specific antibody) and rabbit antinitrotyrosine (a 'footprint' of a reactive form of nitric oxide [peroxynitritel). RESULTS: Levels of CEC were significantly higher in patients with active SLE compared with those in healthy controls (mean +/- SEM 32+/-7/ml versus 5+/-2/ml; P = 0.0028) and were correlated positively with plasma C3a in these patients (r = 0.81, P = 0.0008). Furthermore, CEC from these patients expressed an activated phenotype, as indicated by staining for nitrotyrosine. CONCLUSION: Elevated levels of CEC observed in patients with active SLE may represent a marker of endothelial injury. The activated phenotype of these cells suggests that they may be capable of further potentiating vascular injury by the production of inflammatory and prothrombotic mediators and engaging in heterotypic aggregation with neutrophils or platelets
— id: 20665, year: 2001, vol: 44, page: 1203, stat: Journal Article,

Interaction between 52 kDa SSA/Ro and deubiquitinating enzyme UnpEL: a clue to function
Di Donato F; Chan EK; Askanase AD; Miranda-Carus M; Buyon JP
2001 Sep;33(9):924-934, International journal of biochemistry & cell biology
The detection of isolated heart block in utero strongly predicts the presence of maternal autoantibodies reactive with 52 kDa SSA/Ro. The mechanisms that underlie this observation may be elucidated by defining the function of the target antigen. The initial approach was to identify proteins interactive with 52Ro using transcriptional activity in the yeast 2-hybrid system. A cDNA library was constructed using RNA isolated from human fetal hearts (12-23 weeks) and cloned into the HybriZAP vector encoding the activation domain of GAL4(AD) as target. Approximately 7x10(6) cDNAs were cotransformed with the bait into YRG-2. Plasmids from five interactive colonies were sequenced and three identified as the specific human deubiquitinating enzyme, UnpEL. UnpEL did not interact with bait plasmid encoding 52beta, an alternative leucine zipper-minus form of 52 kDa SSA/Ro which is maximally expressed in fetal life. The mammalian 2-hybrid assay confirmed the interaction between full-length 52Ro and UnpEL. Further support for a biologic interaction was the marked redistribution in cellular localization of UnpEL following cotransfection of the two proteins into cultured human cardiocytes, human renal carcinoma cells (293 cells), and monkey kidney fibroblasts (COS-1). In conclusion, the interaction of full-length 52Ro and UnpEL implies that the former may also be involved in the ubiquitin pathway, an observation of particular interest since 52Ro contains a RING finger domain, a motif common to several recently reported proteins involved in modulating ubiquitination. The absence of an interaction with 52beta raises the consideration that regulation of protein ubiquitination might differ in fetal life
— id: 21112, year: 2001, vol: 33, page: 924, stat: Journal Article,

High thrombosis rate after fetal loss in antiphospholipid syndrome: effective prophylaxis with aspirin
Erkan, D; Merrill, J T; Yazici, Y; Sammaritano, L; Buyon, J P; Lockshin, M D
2001 Jun;44(6):1466-1467, Arthritis & rheumatism
— id: 73546, year: 2001, vol: 44, page: 1466, stat: Journal Article,

Congenital heart block: development of late-onset cardiomyopathy, a previously underappreciated sequela
Moak, J P; Barron, K S; Hougen, T J; Wiles, H B; Balaji, S; Sreeram, N; Cohen, M H; Nordenberg, A; Van Hare, G F; Friedman, R A; Perez, M; Cecchin, F; Schneider, D S; Nehgme, R A; Buyon, J P
2001 Jan;37(1):238-242, Journal of the American College of Cardiology
OBJECTIVE: We report 16 infants with complete congenital heart block (CHB) who developed late-onset dilated cardiomyopathy despite early institution of cardiac pacing. BACKGROUND: Isolated CHB has an excellent prognosis following pacemaker implantation. Most early deaths result from delayed initiation of pacing therapy or hemodynamic abnormalities associated with congenital heart defects. METHODS: A multi-institutional study was performed to identify common clinical features and possible risk factors associated with late-onset dilated cardiomyopathy in patients born with congenital CHB. RESULTS: Congenital heart block was diagnosed in utero in 12 patients and at birth in four patients. Ten of 16 patients had serologic findings consistent with neonatal lupus syndrome (NLS). A pericardial effusion was evident on fetal ultrasound in six patients. In utero determination of left ventricular (LV) function was normal in all. Following birth, one infant exhibited a rash consistent with NLS and two had elevated hepatic transaminases and transient thrombocytopenia. In the early postnatal period, LV function was normal in 15 patients (shortening fraction [SF] = 34 +/- 7%) and was decreased in one (SF = 20%). A cardiac pacemaker was implanted during the first two weeks of life in 15 patients and at seven months in one patient. Left ventricular function significantly decreased during follow-up (14 days to 9.3 years, SF = 9% +/- 5%). Twelve of 16 patients developed congestive heart failure before age 24 months. Myocardial biopsy revealed hypertrophy in 11 patients, interstitial fibrosis in 11 patients, and myocyte degeneration in two patients. Clinical status during follow-up was guarded: four patients died from congestive heart failure; seven required cardiac transplantation; one was awaiting cardiac transplantation; and four exhibited recovery of SF (31 +/- 2%). CONCLUSIONS: Despite early institution of cardiac pacing, some infants with CHB develop LV cardiomyopathy. Patients with CHB require close follow-up not only of their cardiac rate and rhythm, but also ventricular function
— id: 73547, year: 2001, vol: 37, page: 238, stat: Journal Article,

Leucine zipper domain of 52 kDa SS-A/Ro promotes protein dimer formation and inhibits in vitro transcription activity
Wang D; Buyon JP; Yang Z; Di Donato F; Miranda-Carus ME; Chan EK
2001 Dec 5;1568(2):155-161, Biochimica & biophysica acta
Two forms of the human 52 kDa SS-A/Ro protein autoantigen, 52alpha and 52beta, are products of alternative mRNA splicing. The 52alpha form is ubiquitously expressed whereas 52beta, lacking the central leucine zipper domain, has been detected at higher levels than 52alpha during certain stages of fetal development. Because 52alpha has sequence similarity with macromolecules associated with transcriptional regulation and the two forms differ only in that 52beta does not contain the leucine zipper, their roles in protein dimer formation and in transcriptional activity were examined. Employing the yeast two-hybrid system, 52alpha was shown to interact with itself but not 52beta. The homodimerization of 52alpha was independently confirmed in gel filtration chromatography using in vitro cDNA template derived translation products and in HL-60 cell extracts; two peaks were observed corresponding to dimer and monomer of 52alpha, while in vitro the translation product of 52beta exhibited only a single monomer peak. In addition, dimer formation was also demonstrated in a chemical cross-linking experiment using HeLa cells transfected with 52alpha. To evaluate effects on transcription, eukaryotic expression plasmids encoding 52alpha or 52beta fused with the GAL4 DNA binding (DB) domain were co-transfected into 293 cells together with a luciferase reporter vector. A 6-fold increase in transcription activity of the reporter was detected with the GAL4-DB-52beta fusion constructs compared to GAL4-DB-52alpha or the empty vector control. We speculate that the ratio of cellular 52alpha and 52beta may play an important role in regulating gene expression as potential repressor and activator respectively
— id: 32696, year: 2001, vol: 1568, page: 155, stat: Journal Article,

Clinical trials in systemic lupus erythematosus
Buyon JP
2000 Feb;2(1):11-12, Current rheumatology reports
— id: 39501, year: 2000, vol: 2, page: 11, stat: Journal Article,

The heart and skin of neonatal lupus: does maternal health matter?
Buyon, J P
2000 Jun 15;108(9):741-743, American journal of medicine
— id: 73548, year: 2000, vol: 108, page: 741, stat: Journal Article,

Pulsed Doppler echocardiographic assessment of the fetal PR interval
Glickstein, JS; Buyon, J; Friedman, D
2000 JUL 15 ;86(2):236-239, American journal of cardiology
— id: 54601, year: 2000, vol: 86, page: 236, stat: Journal Article,

Expression of SS-A/Ro and SS-B/La antigens in skin biopsy specimens of patients with photosensitive forms of lupus erythematosus
Ioannides D; Golden BD; Buyon JP; Bystryn JC
2000 Mar;136(3):340-346, Archives of dermatology
CONTEXT: The reason that only some patients with lupus erythematosus (LE) develop autoantibodies to SS-A/Ro and SS-B/La antigens and photosensitivity is unknown. One hypothesis is that both events are related to the level of expression of these antigens in the skin. OBJECTIVE AND DESIGN: To test this hypothesis, we measured the expression of the 52-kd SS-A/Ro, 60-kd SS-A/Ro, and 48-kd SS-B/La antigens in normal sun-protected and sun-exposed skin in 14 patients with LE with photosensitivity, 12 patients with LE without photosensitivity, and 4 normal individuals. The presence of circulating antibodies to these antigens was measured in all patients. SETTING: Outpatient clinic in an academic medical center. RESULTS: We found that the expression of the 52-kd SS-A/Ro, 60-kd SS-A/Ro, and 48-kd SS-B/La antigens in skin biopsy specimens obtained from the same site was 4- to 10-fold higher in patients with LE with photosensitivity than in those patients with LE without photosensitivity (P<.001). Antigen expression was highly correlated with the presence and titer of circulating anti-SS-A/Ro and anti-SS-B/La antibodies (P<.001). CONCLUSIONS: These findings indicate that photosensitivity and the presence and titer of circulating anti-SS-A/Ro and anti-SS-B/La antibodies are both directly correlated with the expression of accessible and immunoreactive SS-A/Ro and SS-B/La antigens in the skin specimens of patients with LE. Thus, the expression of these antigens in keratinocytes may be an important determinant of the development of both SS-A/Ro and SS-B/La autoantibodies and of photosensitive forms of LE
— id: 16219, year: 2000, vol: 136, page: 340, stat: Journal Article,

Anti-SSA/Ro and anti-SSB/La autoantibodies bind the surface of apoptotic fetal cardiocytes and promote secretion of TNF-alpha by macrophages
Miranda-Carus ME; Askanase AD; Clancy RM; Di Donato F; Chou TM; Libera MR; Chan EK; Buyon JP
2000 Nov 1;165(9):5345-5351, Journal of immunology
Despite the near universal association of congenital heart block and maternal Abs to SSA/Ro and SSB/La, the intracellular location of these Ags has made it difficult to substantiate their involvement in pathogenicity. To define whether components of the SSA/Ro-SSB/La complex, which translocate during apoptosis, are indeed accessible to extracellular Abs, two approaches were taken: immunoprecipitation of surface biotinylated proteins and scanning electron microscopy. Human fetal cardiocytes from 16-24-wk abortuses were cultured and incubated with staurosporine to induce apoptosis. Surface biotinylated 48-kDa SSB/La was reproducibly immunoprecipitated from apoptotic, but not nonapoptotic cardiocytes. Surface expression of SSA/Ro and SSB/La was further substantiated by scanning electron microscopy. Gold particles (following incubation with gold-labeled sera containing various specificities of anti-SSA/Ro-SSB/La Abs and murine mAb to SSB/La and 60-kDa SSA/Ro) were consistently observed on early and late apoptotic cardiocytes. No particles were seen after incubation with control antisera. To evaluate whether opsonized apoptotic cardiocytes promote inflammation, cells were cocultured with macrophages. Compared with nonapoptotic cardiocytes or apoptotic cardiocytes incubated with normal sera, apoptotic cardiocytes preincubated with affinity-purified Abs to SSB/La, 52-kDa SSA/Ro, or 60-kDa SSA/Ro increased the secretion of TNF-alpha from cocultured macrophages. In summary, apoptosis results in surface accessibility of all SSA/Ro-SSB/La Ags for recognition by circulating maternal Abs. It is speculated that in vivo such opsonized apoptotic cardiocytes promote an inflammatory response by resident macrophages with damage to surrounding conducting tissue
— id: 32697, year: 2000, vol: 165, page: 5345, stat: Journal Article,

Cutaneous manifestations of neonatal lupus without heart block: characteristics of mothers and children enrolled in a national registry
Neiman AR; Lee LA; Weston WL; Buyon JP
2000 Nov;137(5):674-680, Journal of pediatrics
OBJECTIVE: To extend the information base on cutaneous manifestations of neonatal lupus erythematosus (NLE) with regard to maternal disease, sex of child, onset, localization, influence of UV light, prognosis, and recurrence rates in subsequent pregnancies. METHODS: Review of records from the Research Registry for Neonatal Lupus. RESULTS: The cohort includes 47 mothers (83% white) whose sera contain anti-SSA/Ro, anti-SSB/La, and/or anti-U1-ribonucleoprotein antibodies and their 57 infants (20 boys and 37 girls) diagnosed with cutaneous NLE (absent heart disease) between 1981 and 1997. At detection of the child's rash, 13 mothers were asymptomatic, 11 had an undifferentiated autoimmune syndrome (UAS), 9 had systemic lupus erythematosus (SLE), 7 Sjogren's syndrome (SS), 6 SLE/SS, and 1 rheumatoid arthritis/SS; 20 reported photosensitivity. Within 5 years, 7 asymptomatic mothers experienced disease progression: 1 developed photosensitivity, 2 SLE, 3 SS, 1 SLE/SS; in 2 mothers UAS progressed to SLE; and 2 mothers with SS developed SLE. The infant's rash often followed UV light exposure; mean age at detection was 6 weeks, and mean duration was 17 weeks. All had facial involvement (periorbital region most common) followed by the scalp, trunk, extremities, neck, and intertriginous areas. In 37, the rash resolved without sequelae, 43% of which were untreated. A quarter had residual sequelae that included telangiectasia and dyspigmentation. One child developed Hashimoto's thyroiditis, and 2 developed systemic-onset juvenile rheumatoid arthritis. Of 20 subsequent births, 7 children were healthy, 2 had congenital heart block (CHB) only, 4 CHB and skin rash, and 7 skin rash only. CONCLUSIONS: Future pregnancies should be monitored by serial echocardiograms, given the substantial risk for heart block. Affected children should be observed for later development of a rheumatic disease
— id: 39434, year: 2000, vol: 137, page: 674, stat: Journal Article,

Fourth international workshop on neonatal lupus syndromes and the Ro/SSA-La/SSB System
Brucato, A; Buyon, J P; Horsfall, A C; Lee, L A; Reichlin, M
1999 Jan-Feb;17(1):130-136, Clinical & experimental rheumatology
— id: 73551, year: 1999, vol: 17, page: 130, stat: Journal Article,

Assessing disease activity in SLE patients during pregnancy
Buyon JP; Kalunian KC; Ramsey-Goldman R; Petri MA; Lockshin MD; Ruiz-Irastorza G; Khamashta M
1999 ;8(8):677-684, Lupus
— id: 11927, year: 1999, vol: 8, page: 677, stat: Journal Article,

Congenital heart block: Autoimmune associated or auto-immune mediated?
Buyon, JP
1999 MAY-JUN ;7(3):98-104, Immunologist
The association of congenital heart block with maternal autoantibodies to SSA/Ro and/or SSB/La ribonucleoproteins suggests a direct role for the autoantibodies in tissue injury. Here, Jill P. Buyon discusses possible pathological mechanisms, including the nature of the target antigen,the role of autoantibodies in altering L-type calcium channel function within the heart, and the potential role of apoptosis in making target antigens available to autoantibodies
— id: 54009, year: 1999, vol: 7, page: 98, stat: Journal Article,

Equivalence trials in SLE research: issues to consider
Kim MY; Buyon JP; Petri M; Skovron ML; Shore RE
1999 ;8(8):620-626, Lupus
In contrast to the objective of most clinical trials, which is to demonstrate superiority of an experimental treatment over a standard or placebo, the aim of an equivalence trial is to show that two treatments are equivalent in outcome or only marginally different. This would be of interest when an experimental treatment offers advantages such as reduced toxicity, ease of administration, or cost relative to the standard. Demonstrating equivalence may also be a goal when evaluating the safety of certain drugs because similarity in the risks of an adverse event in subjects exposed and unexposed to the drug is an indication of its safety. The classical formulation of the null hypothesis of treatment equality that is used in superiority trials is not applicable to equivalence trials because absolute equivalence between treatment groups cannot be proven. The strategy in equivalence trials is to define a maximum difference between treatment groups that is clinically acceptable and then assess whether there is sufficient evidence from the trial to conclude that the true treatment difference is within this acceptable range. In this paper, we discuss issues surrounding the planning, conduct, and analysis of equivalence trials in the context of SLE, with examples from the SELENA study
— id: 11928, year: 1999, vol: 8, page: 620, stat: Journal Article,

Classification and definition of major flares in SLE clinical trials
Petri, M; Buyon, J; Kim, M
1999 ;8(8):685-691, Lupus
Multiple reliable and valid disease activities indices exist and have been used successfully in longitudinal studies. However, the definition of flare, using these intruments, has not been universally decided or accepted. Because flare is one of the three major patterns of lupus activity, it will remain an important outcome measure in both longitudinal and clinical trial studies
— id: 132527, year: 1999, vol: 8, page: 685, stat: Journal Article,

Comparison of treatment with fluorinated glucocorticoids to the natural history of autoantibody-associated congenital heart block: retrospective review of the research registry for neonatal lupus
Saleeb S; Copel J; Friedman D; Buyon JP
1999 Nov;42(11):2335-2345, Arthritis & rheumatism
OBJECTIVE: To compare intervention with fluorinated glucocorticoids to the natural history of untreated congenital heart block (CHB) with respect to conduction abnormalities, associated effusions, ascites, and hydrops fetalis, and the requirement for a pacemaker. METHODS: Records of all mothers enrolled in the Research Registry for Neonatal Lupus were reviewed. The cohort includes 47 mothers whose sera contain anti-SSA/Ro or anti-SSB/La antibodies, and their 50 offspring with CHB, in whom at least 4 echocardiograms were performed after in utero diagnosis. In 28 pregnancies, mothers received dexamethasone 4-9 mg/day for 3-19 weeks or betamethasone 12-24 mg/week for >6 weeks (group A). In 22 pregnancies, fluorinated steroids were not used (group B). RESULTS: Third-degree block was present in 21 fetuses in group A and 18 fetuses in group B; none were reversible despite steroid treatment. Three fetuses in group A and 2 in group B progressed from second-degree block, alternating with third-degree block, to permanent third-degree block at birth and postnatally. Of 4 fetuses in group A with second-degree block at presentation, all reverted to first-degree block by birth; 2 remain so at age 4 years, 1 alternates between first-degree and second-degree block at 2 years, and the fourth is in second-degree block at age 4 years. Of 2 fetuses in group B with second-degree block at presentation, both progressed to permanent third-degree block postnatally. Initial echocardiographic evaluation revealed pericardial effusions in 13 group A versus 4 group B fetuses, pleural effusions in 2 group A versus 0 group B, ascites in 8 group A versus 0 group B (P < 0.007), hydrops fetalis in 8 group A versus 0 group B (P < 0.007), and intrauterine growth restriction in 1 group A versus 1 group B. Pericardial effusions resolved and reappeared in both groups. Steroid therapy was most effective in the resolution of pleural effusions (2 of 2), ascites (6 of 8), and hydrops fetalis (5 of 8). Oligohydramnios ensued in 9 group A and 2 group B fetuses. Although fetuses in group A had more complications at presentation than those in group B, there were no significant differences in the duration of pregnancy (35.7 weeks versus 37.0 weeks), the number of deaths (4 versus 1), final degree of heart block, or requirement for a pacemaker (14 versus 11). CONCLUSION: While prospective trials are needed, these data suggest that fluorinated steroids should be considered for fetuses with incomplete block or hydropic changes. Serial echocardiograms are recommended to monitor fetal progress. It remains to be determined whether third-degree block is reversible if therapy is initiated immediately upon detection
— id: 11933, year: 1999, vol: 42, page: 2335, stat: Journal Article,

mRNA and protein expression of SSA/Ro and SSB/La in human fetal cardiac myocytes cultured using a novel application of the Langendorff procedure
Tseng CE; Miranda E; Di Donato F; Boutjdir M; Rashbaum W; Chan EK; Buyon JP
1999 Feb;45(2):260-269, Pediatric research
Irreversible congenital heart block (CHB) and the transient rash of neonatal lupus are strongly associated with maternal antibodies to SSA/Ro and SSB/La proteins; however, the precise mechanism by which these antibodies mediate organ-specific injury is not yet defined. Culturing of keratinocytes has provided critical insights. Accordingly, successful culturing of human fetal cardiac myocytes at high yield would constitute a powerful tool to directly examine conditions that promote expression of the target autoantigens. To accomplish this aim, fetal cardiac myocytes from 18- to 22-wk abortuses were established in culture using a novel technique in which cells were isolated after perfusion of the aorta with collagenase in a Langendorff apparatus. After preplating to decrease fibroblast contamination, cardiocytes were grown in flasks and slide chambers. Staining with monoclonal anti-sarcomeric alpha-actinin revealed the expected striations typical of cardiac myocytes in 70-90% of the cells after 4 d in culture. Furthermore, the cells were observed to beat at rates varying between 25-75 beats per minute (bpm) after the addition of 1.8 mM CaCl2. An average yield of 45-60 x 10(6) cells was obtained from a 3- to 5-g heart. Cellular localization of SSA/Ro and SSB/La by indirect immunofluorescence and demonstration of mRNA expression by reverse transcriptase polymerase chain reaction supports the feasibility of cultured cardiac myocytes for the study of congenital heart block. In contrast to the increased expression of SSA/Ro reported for keratinocytes, incubation of cultured human cardiac myocytes with either 17beta-estradiol or progesterone did not alter mRNA expression or cellular localization of 48 kD SSB/La, 52 kD SSA/Ro, or 60 kD SSA/Ro. In summary, we describe a novel method to successfully culture human fetal cardiac myocytes that should provide a valuable resource for investigation of the molecular mechanism(s) contributing to the development of congenital heart block. Differential constitutive and estradiol-induced expression of 52 and 60 kD SSA/Ro in human cardiac myocytes compared with keratinocytes may be a factor contributing to the marked discordance of clinically detectable injury in these two target tissues
— id: 57078, year: 1999, vol: 45, page: 260, stat: Journal Article,

Defining a novel 75-kDa phosphoprotein associated with SS-A/Ro and identification of distinct human autoantibodies
Wang, D; Buyon, J P; Zhu, W; Chan, E K
1999 Nov;104(9):1265-1275, Journal of clinical investigation
Mothers of children with neonatal lupus erythematosus (NLE) and heart block, as well as patients with Sjogren's syndrome (SS) and systemic lupus erythematosus, have serum autoantibodies that recognize SS-A/Ro autoantigens including the 60-kDa ribonucleoprotein. By yeast 2-hybrid screening, we identified a novel 75-kDa protein (pp75) that interacts with the carboxyl 70% of 60-kDa SS-A/Ro. The specificity of interaction was confirmed using mammalian 2-hybrid and chemical crosslinking studies. Immunoprecipitation with radiolabeled HeLa cell extracts showed that pp75 was phosphorylated and associated with 2 other phosphoproteins of 64 kDa. In Northern blot analysis, pp75 was expressed in all tissues analyzed; the highest expression was in the human heart. Based on immunofluorescence of transfected HeLa cells, pp75 is localized predominantly in the cytoplasm, an observation confirmed by immunohistochemistry in untransfected cells. Based on Western blot and ELISA assays, sera from 14 of 84 mothers of children with NLE recognized pp75, including 1 mother in whom anti-SS-A/Ro antibodies were not detected. In addition, sera from 5 of 80 patients with SS were positive for anti-pp75 antibody. Identification of molecular partners is a first step toward elucidating the functions and possible involvement in pathogenesis of long-recognized autoantigens such as 60-kDa SS-A/Ro, which are at present poorly understood
— id: 73549, year: 1999, vol: 104, page: 1265, stat: Journal Article,

The prevalence of cardiomyopathy observed in patients recorded in the research registry for neonatal lupus
Abbot, AE; Buvon, JP; Friedman, D
1998 SEP ;41(9):S262-S262, Arthritis & rheumatism
— id: 53746, year: 1998, vol: 41, page: S262, stat: Journal Article,

Serum and immunoglobulin G from the mother of a child with congenital heart block induce conduction abnormalities and inhibit L-type calcium channels in a rat heart model
Boutjdir M; Chen L; Zhang ZH; Tseng CE; El-Sherif N; Buyon JP
1998 Jul;44(1):11-19, Pediatric research
Although a strong clinical association exists between congenital heart block (CHB) and an immune response to SSA/Ro and SSB/La proteins, a causative role of these antibodies in the pathogenesis is just emerging. In a preliminary report, we have demonstrated that IgG fractions isolated from the sera of mothers whose children have CHB are arrhythmogenic in the human fetal heart. To more precisely define the arrhythmogenic effect of anti-SSA/Ro-SSB/La antibodies, we used the readily available rat heart model to record: 1) ECGs from Langendorff beating hearts; 2) action potentials from atrioventricular (AV) nodal preparations; 3) L-type Ca currents, I(Ca) at the whole-cell and single channel levels; and 4) other currents such as the transient outward K+ current, I(to), the inward rectifier K+ current, I(K1), and the Na+ current, I(Na). Perfusion of hearts with purified IgG (800 microg/mL), isolated from the serum of a mother with SSA/Ro and SSB/La antibodies whose child had CHB, resulted in bradycardia associated with 2:1 AV block. Simultaneous action potentials were recorded from dissected atrial and AV nodal areas of the rat heart. Superfusion of these preparations with the same mother's IgG fraction resulted in 2:1 AV block followed by complete inhibition of AV nodal action potential. Because AV nodal electrogenesis is largely dependent on I(Ca), the effect of these antibodies on I(Ca) was subsequently determined. Superfusion of myocytes with whole serum or purified IgG (80 microg/mL) from the same mother consistently inhibited whole cell I(Ca), ensemble average Ba2+ currents (I(Ba)) and open state probability, p(o), without affecting the channel conductance. IgG had no significant effect on I(to), I(K1), or I(Na). Whole sera and IgG fractions from a healthy mother with no detectable anti-SSA/Ro or SSB/La antibodies did not inhibit I(Ca) or I(Ba). These results demonstrate that IgG containing anti-SSA/Ro and -SSB/La antibodies induces complete AV block in beating hearts and in multicellular preparations, thus implicating a preferential interaction of these autoantibodies with Ca channels and/or associated regulatory proteins. This is consistent with the observed inhibition of Ca channels that may be a critical factor contributing to the pathogenesis of CHB
— id: 12093, year: 1998, vol: 44, page: 11, stat: Journal Article,

Hormone replacement therapy in postmenopausal women with systemic lupus erythematosus
Buyon JP
1998 Winter;53(1):13-17, Journal of the American Medical Womens Association (1972)
Given the female preponderance of systemic lupus erythematosus (SLE) in humans, the adverse effects of female gender and sex hormones in murine lupus, and numerous reports (retrospective, often anecdotal and uncontrolled) describing a temporal association between estrogen exposure and development or exacerbation of SLE, it is easy to accept that estrogens and SLE simply do not mix. While there are valid concerns regarding the use of exogenous estrogens in women with SLE, there are also potential health benefits to be considered. Several salutary effects of postmenopausal estrogens assume particular importance in SLE where the risks of osteoporosis, exaggerated by menopause (natural or cyclophosphamide-induced) and glucocorticoids, are substantial. Moreover, hormone replacement therapy (HRT) is associated with a 40% reduction in the risk of coronary artery disease, higher levels of high-density lipoprotein-cholesterol, and decreased levels of low-density lipoprotein-cholesterol and plasminogen-activator inhibitor type 1, benefits that should be especially applicable to post-menopausal women with SLE. More recent studies, albeit retrospective and absent the use of validated measures of disease activity, suggest that HRT may be well tolerated. In counseling patients regarding lupus and pregnancy, there are now clinical predictors of pregnancy outcome, and patients in remission tend to have good outcomes. The same principles may be true regarding advice on the use of HRT; patients with inactive or stable/moderate disease and at low risk for thrombosis may benefit without a change in lupus activity. Large prospective double-blind placebo controlled studies inclusive of all ethnic groups such as the Safety of Estrogens in Lupus Erythematosus-National Assessment (SELENA) trial should provide the basis for definitive recommendations
— id: 57109, year: 1998, vol: 53, page: 13, stat: Journal Article,

Neonatal lupus and autoantibodies reactive with SSA/Ro-SSB/La
Buyon JP
1998 ;107:23-30, Scandinavian journal of rheumatology. Supplement
— id: 12067, year: 1998, vol: 107, page: 23, stat: Journal Article,

The effects of pregnancy on autoimmune diseases
Buyon JP
1998 Mar;63(3):281-287, Journal of leukocyte biology
Internal gestation of a genetically foreign conceptus challenges the maternal host to circumvent immunological processes that recognize and eliminate nonself molecules. Accordingly, human viviparity involves a wide range of immunological modifications. This review examines the relationship between pregnancy and several autoimmune diseases prevalent in women. Pregnancy is associated with improvement in the clinical signs and symptoms of rheumatoid arthritis in more than 70% of patients. Maternal-fetal disparity in alleles of HLA-DRbeta1, DQalpha, and DQbeta has been reported to be associated with pregnancies characterized by remission or improvement. These observations suggest that presentation of fetal DQalpha peptides might correct autoimmunity in patients with RA either by induction of maternal-regulatory T cells, or by affecting the maternal T cell receptor repertoire. In contrast, the course of systemic lupus erythematosus is more variable. Whether flare rates increase during or after pregnancy is unsettled, since individual patient series vary in the characteristics of patients accepted for study and in definitions of flare. Despite a high overall flare rate in some series approaching 60%, recorded flares were usually not severe. Only limited data are available regarding the incidence or outcome for either the mother with scleroderma or her fetus. The extent of diffuse skin disease and systemic involvement, particularly pulmonary, cardiac and renal, may be more important than the duration of the disease; limited disease carries a better prognosis for the mother and fetus. Highly specific autoantibody profiles in the mother (independent of whether she has a clinical disease) are associated with fetal demise and neonatal lupus syndromes, the most serious manifestation of which is isolated congenital heart block. The former is associated with antiphospholipid antibodies and the latter with antibodies directed against SSA/Ro and SSB/La polypeptides
— id: 12154, year: 1998, vol: 63, page: 281, stat: Journal Article,

Autoimmune-associated congenital heart block: demographics, mortality, morbidity and recurrence rates obtained from a national neonatal lupus registry
Buyon JP; Hiebert R; Copel J; Craft J; Friedman D; Katholi M; Lee LA; Provost TT; Reichlin M; Rider L; Rupel A; Saleeb S; Weston WL; Skovron ML
1998 Jun;31(7):1658-1666, Journal of the American College of Cardiology
OBJECTIVES: The present study describes the demographics, mortality, morbidity and recurrence rates of autoimmune-associated congenital heart block (CHB) using information from the Research Registry for Neonatal Lupus. BACKGROUND: Isolated CHB detected at or before birth is strongly associated with maternal autoantibodies to 48-kD SSB/La, 52-kD SSA/Ro and 60-kD SSA/Ro ribonucleoproteins and is a permanent manifestation of the neonatal lupus syndromes (NLS). Available data are limited by the rarity of the disease. RESULTS: The cohort includes 105 mothers whose sera contain anti-SSA/Ro or anti-SSB/La antibodies, or both, and their 113 infants diagnosed with CHB between 1970 and 1997 (56 boys, 57 girls). Of 87 pregnancies in which sufficient medical records were available, bradyarrhythmia confirmed to be CHB was initially detected before 30 weeks of gestation in 71 (82%) (median time 23 weeks). There were no cases in which major congenital cardiac anatomic defects were considered causal for the development of CHB; in 14 there were minor abnormalities. Twenty-two (19%) of the 113 children died, 16 (73%) within 3 months after birth. Cumulative probability of 3-year survival was 79%. Sixty-seven (63%) of 107 live-born children required pacemakers: 35 within 9 days of life, 15 within 1 year, and 17 after 1 year. Forty-nine of the mothers had subsequent pregnancies: 8 (16%) had another infant with CHB and 3 (6%) had a child with an isolated rash consistent with NLS. CONCLUSIONS: Data from this large series substantiate that autoantibody-associated CHB is not coincident with major structural abnormalities, is most often identified in the late second trimester, carries a substantial mortality in the neonatal period and frequently requires pacing. The recurrence rate of CHB is at least two- to three-fold higher than the rate for a mother with anti-SSA/Ro-SSB/La antibodies who never had an affected child, supporting close echocardiographic monitoring in all subsequent pregnancies, with heightened surveillance between 18 and 24 weeks of gestation
— id: 12110, year: 1998, vol: 31, page: 1658, stat: Journal Article,

Perinatal monitoring of fetal well-being in the presence of congenital heart block
Friedman, D M; Zervoudakis, I; Buyon, J P
1998 ;15(12):669-673, American journal of perinatology
We present a case of congenital complete heart block associated with maternal autoantibodies in which a normal labor and delivery could safely be allowed to proceed despite the absence of the usual tool of electronic fetal heart rate monitoring for fetal distress, by the technique of rupturing membranes and using a fetal scalp electrode electrocardiographic tracing to assess the fetal atrial rate
— id: 73550, year: 1998, vol: 15, page: 669, stat: Journal Article,

Human anticardiolipin monoclonal autoantibodies cause placental necrosis and fetal loss in BALB/c mice
Ikematsu, W; Luan, F L; La Rosa, L; Beltrami, B; Nicoletti, F; Buyon, J P; Meroni, P L; Balestrieri, G; Casali, P
1998 Jun;41(6):1026-1039, Arthritis & rheumatism
OBJECTIVE: To analyze the structure, specificity, and in vivo pathogenetic potential of 2 human anticardiolipin (aCL) monoclonal antibodies (MAb). METHODS: Human aCL IgG MAb were generated from hybridized Epstein-Barr virus-induced B cell lines from a healthy subject (MAb 519) and from a patient with primary antiphospholipid syndrome (MAb 516). Studies of antigen-binding specificity and analysis of Ig V-gene mutations were carried out. The MAb were independently injected into mated female BALB/c mice, and their effect on pregnancy outcome was compared with that of MAb 57, a highly mutated and antigen-selected human IgG1lambda rabies virus antibody. RESULTS: Both MAb 519 and MAb 516 utilized minimally mutated V(H)DJ(H) and VkappaJkappa gene segments and bound cardiolipin and other anionic phospholipids in the absence of beta2-glycoprotein I (beta2-GPI). The mice injected with aCL MAb displayed a significantly higher rate of fetal resorption and a significant reduction in fetal and placental weight as compared with those injected with MAb 57. These findings were accompanied by a finding of placental human IgG deposition and necrosis in the aCL MAb-treated animals. CONCLUSION: The results of this study indicate that human aCL IgG that are beta2-GPI independent can induce pathology
— id: 73552, year: 1998, vol: 41, page: 1026, stat: Journal Article,

The expression of Ro and La antigens is increased in the skin of patients with photosensitive lupus erythematosus
Ioannides, D; Golden, B; Buyon, J; Bystryn, JC
1998 APR ;110(4):635-635, Journal of investigative dermatology
— id: 53527, year: 1998, vol: 110, page: 635, stat: Journal Article,

Decreased anticoagulant protein S and the homologous sex hormone binding globulin in patients enrolled in the SELENA (estrogen safety) study
Merrill, JT; Schappert, J; Shrikv, RC; Zhang, HW; Petrie, M; Buyon, JP
1998 SEP ;41(9):S139-S139, Arthritis & rheumatism
— id: 53738, year: 1998, vol: 41, page: S139, stat: Journal Article,

Accessibility of SSA/Ro and SSB/La antigens to maternal autoantibodies in apoptotic human fetal cardiac myocytes
Miranda ME; Tseng CE; Rashbaum W; Ochs RL; Casiano CA; Di Donato F; Chan EK; Buyon JP
1998 Nov 1;161(9):5061-5069, Journal of immunology
Access of intracellular Ags SSA/Ro and SSB/La to cognate maternal autoantibodies is unexplained despite their strong association with congenital heart block. To investigate the hypothesis that apoptosis facilitates surface accessibility of these Ags, human fetal cardiac myocytes from 16- to 22-wk abortuses were established in culture using a novel technique in which cells were isolated after perfusing the aorta with collagenase. Confirmation of cardiac myocytes included positive staining with antisarcomeric alpha-actinin and contractility induced by 1.8 mM calcium. Incubation with 0.5 microM staurosporine or 0.3 mM 2,3-dimethoxy-1,4-naphthoquinone induced the characteristic morphologic and biochemical changes of apoptosis. The cellular topology of Ro and La was evaluated with confocal microscopy and determined in nonapoptotic and apoptotic cardiocytes by indirect immunofluorescence. In permeabilized nonapoptotic cardiocytes, Ro and La were predominantly nuclear, and propidium iodide (PI) stained the nucleus. In early apoptotic cardiocytes, condensation of the PI- and Ro- or La-stained nucleus was observed, accompanied by Ro/La fluorescence around the cell periphery. In later stages of apoptosis, nuclear Ro and La staining became weaker, and PI demonstrated nuclear fragmentation. Ro/La-stained blebs emerged from the cell membrane, a finding observed in nonpermeabilized cells, supporting an Ab-Ag interaction at the cell surface. In summary, induction of apoptosis in cultured cardiocytes results in surface translocation of Ro/La and recognition by Abs. Although apoptotic cells are programmed to die and do not characteristically evoke inflammation, binding of maternal Abs and subsequent influx of leukocytes could damage surrounding healthy fetal cardiocytes
— id: 7685, year: 1998, vol: 161, page: 5061, stat: Journal Article,

Recognition by maternal autoantibodies of 48kD La, 52kD and 60kDd Ro on the cell surface of apoptotic human fetal cardiocytes: A potential mediator of inflammation
Miranda, E; Chan, EKL; Tseng, C; DiDonato, F; Gordon, T; Rebarber, A; Buvon, JP
1998 SEP ;41(9):S128-S128, Arthritis & rheumatism
— id: 53737, year: 1998, vol: 41, page: S128, stat: Journal Article,

Induction of antibodies reactive with SSA/Ro-SSB/La and development of congenital heart block in a murine model
Miranda-Carus ME; Boutjdir M; Tseng CE; DiDonato F; Chan EK; Buyon JP
1998 Dec 1;161(11):5886-5892, Journal of immunology
To correlate the arrhythmogenic effects of maternal autoantibodies with the genesis of congenital heart block, female BALB/c mice were immunized with human recombinant 48-kDa SSB/La, 60-kDa SSA/Ro, 52-kDa SSA/Ro (52alpha), and 52beta (amino acids 169-245 deleted) as well as with murine recombinant 52-kDa SSA/Ro. Control animals received beta-galactosidase or a polypeptide encoded by pET-28 alone. Following primary immunization and two boosters, high titer responses to the respective Ags were established by ELISA, immunoblotting, and immunoprecipitation. Sera from mice immunized with either human 52alpha or 52beta immunoprecipitated murine 52Ro. mRNA and protein expression of 52Ro was demonstrated in the newborn murine heart. A spectrum of atrioventricular nodal conduction abnormalities was identified by electrocardiogram. First-degree block was detected in 7% of 27 pups born to mothers immunized with 48La, 20% of 54 pups born to 60Ro-immunized mothers, 6% of 56 pups born to 52alpha-immunized mothers, 7% of 86 pups born to 52beta-immunized mothers, and 9% of 22 pups born to mothers immunized with murine 52Ro. Advanced conduction abnormalities were only identified in offspring of 52alpha- or 52beta-immunized mice. In the 52alpha group, one pup had complete block and another had second-degree block (Wenckebach type); in the 52beta group, five pups had complete block. Maternal Abs to the primary immunogens were detected in the pups. No control had any conduction abnormalities. This Ab-specific animal model provides strong evidence for a pathogenic role of anti-SSA/Ro-SSB/La Abs, particularly 52Ro, in the development of congenital heart block. The range and frequency of conduction defects suggest that additional factors promote disease expression
— id: 7686, year: 1998, vol: 161, page: 5886, stat: Journal Article,

Induction of antibodies reactive with SSA/Ro-SSB/La and development of congenital heart block in a murine model
Miranda-Carus, ME; Boutjdir, M; Tseng, C; DiDonato, F; Chan, EKL; Buyon, JP
1998 MAR ;46(3):234A-234A, Journal of investigative medicine
— id: 53504, year: 1998, vol: 46, page: 234A, stat: Journal Article,

Translocation of SSA/Ro and SSB/La in apoptotic human fetal cardiocytes: Pathogenic role in CHB
Miranda-Carus, ME; Tseng, C; Rashbaum, W; Casiano, C; DiDonato, F; Chan, EKL; Buyon, JP
1998 MAR ;46(3):227A-227A, Journal of investigative medicine
— id: 53502, year: 1998, vol: 46, page: 227A, stat: Journal Article,

Isolated cutaneous manifestations of neonatal lupus: Characteristics of mothers and children enrolled in a national registry
Neiman, A; Lee, LA; Buvon, JP
1998 SEP ;41(9):S262-S262, Arthritis & rheumatism
— id: 53745, year: 1998, vol: 41, page: S262, stat: Journal Article,

Differences in pre vs. post-menopausal S
Petri, M; Buyon, J; Kim, M
1998 SEP ;41(9):S67-S67, Arthritis & rheumatism
— id: 53732, year: 1998, vol: 41, page: S67, stat: Journal Article,

Reliability of SELENA SLEDAI and flare as clinical trial outcome measures
Petri, M; Buyon, J; Skovron, ML; Kim, M; Belmont, M; Hahn, B; Kalunian, K; Lahita, R; Lockshin, M; Merrill, J; Sammaritano, L
1998 SEP ;41(9):S218-S218, Arthritis & rheumatism
— id: 53742, year: 1998, vol: 41, page: S218, stat: Journal Article,

Effect of fluorinated glucocorticoids on outcome of autoimmune-associated congenital heart block
Saleeb, S; Copel, J; Friedman, D; Buyon, JP
1998 SEP ;41(9):S80-S80, Arthritis & rheumatism
— id: 53733, year: 1998, vol: 41, page: S80, stat: Journal Article,

Leucine zipper domain of 52kDa SS-A/Ro functions in protein dimerization
Wang, D; Buyon, JP; Chan, EKL
1998 NOV ;9(11):70A-70A, Molecular biology of the cell
— id: 53640, year: 1998, vol: 9, page: 70A, stat: Journal Article,

Arrhythmogenicity of IgG and anti-52-kD SSA/Ro affinity-purified antibodies from mothers of children with congenital heart block
Boutjdir, M; Chen, L; Zhang, Z H; Tseng, C E; DiDonato, F; Rashbaum, W; Morris, A; el-Sherif, N; Buyon, J P
1997 Mar;80(3):354-362, Circulation research
An important advance in the description and understanding of congenital heart block (CHB) came in the 1970s with the observation that mothers of affected infants frequently had autoimmune diseases and, in particular, that many maternal sera contained antibodies to SSA/Ro and SSB/La ribonucleoproteins. Although the molecular biology of the candidate antigens has been extensively defined, the arrhythmogenic and electrophysiological effects of their cognate antibodies on the human fetal heart are unknown. In the present study, we provide evidence that IgG-enriched fractions and anti-52-kD SSA/Ro antibodies affinity-purified from sera of mothers whose children have CHB induce complete atrioventricular (AV) block in the human fetal heart perfused by the Langendorff technique and inhibit L-type Ca2+ currents at the whole-cell and single-channel level. Immunization of female BALB/c mice with recombinant 52-kD SSA/Ro protein generated high-titer antibodies that crossed the placenta during pregnancy and were associated with varying degrees of AV conduction abnormalities, including complete AV block, in the pups. These findings strongly suggest that anti-52-kD SSA/Ro antibodies are causally related to the development of CHB
— id: 73556, year: 1997, vol: 80, page: 354, stat: Journal Article,

Neonatal lupus: long-term outcomes of mothers and children and recurrence rate
Brucato, A; Franceschini, F; Buyon, J P
1997 Sep-Oct;15(5):467-473, Clinical & experimental rheumatology
— id: 73554, year: 1997, vol: 15, page: 467, stat: Journal Article,

Autoantibodies reactive with Ro(SSA) and La(SSB) and pregnancy
Buyon JP
1997 Sep;24 Suppl 50:12-16, Journal of rheumatology
— id: 8095, year: 1997, vol: 24 Suppl 50, page: 12, stat: Journal Article,

Differential phosphorylation of the beta2 integrin CD11b/CD18 in the plasma and specific granule membranes of neutrophils
Buyon JP; Philips MR; Merrill JT; Slade SG; Leszczynska-Piziak J; Abramson SB
1997 Mar;61(3):313-321, Journal of leukocyte biology
Neutrophil aggregation is mediated by the beta2 integrin CD11b/CD18, which has limited expression on the surface membrane of resting cells but is recruited from intracellular organelles after cell activation. We have previously found that CD11b/CD18 newly translocated to the plasma membrane does not contribute to adhesion but must be modified to be functional. Because neutrophil aggregation induced by phorbol myristate acetate (PMA) is accompanied by de novo phosphorylation of the CD18 cytoplasmic tail, we sought to determine whether CD11b/CD18 phosphorylation is separately regulated in the different cellular compartments. Accordingly, [32P]-labeled CD11b/CD18 was immunoprecipitated from purified neutrophil-specific granule or plasma membrane lysates. In plasma membrane fractions, as in whole cell lysates, CD18 became phosphorylated in cells exposed to PMA but not in untreated cells or cells treated with N-formyl-methionyl-leucyl-phenylalanine (fMLP). The alpha chain, CD11b, was phosphorylated under all conditions. In contrast, only marginal phosphorylation of specific granule-associated CD18 or CD11b was observed. Calyculin A, an inhibitor of serine/threonine phosphatases (pp1 > pp2a), induced strong phosphorylation of CD18 in the plasma membrane but not in the specific granules. Addition of intact specific granule membranes to the plasma membranes from PMA-treated neutrophils markedly decreased phosphorylation in both CD11b and CD18 subunits. These data suggest that the phosphorylation of CD11b/CD18, which accompanies neutrophil activation, is limited to plasma membrane-associated molecules. Phosphorylation, either constitutive or induced, is absent in the specific granule membranes. The difference may be due to a specific granule-associated phosphatase, probably distinct from ppl. Therefore adhesion-competent plasma membrane CD11b/CD18 and adhesion-incompetent specific granule CD11b/CD18 differ in their state of phosphorylation
— id: 9735, year: 1997, vol: 61, page: 313, stat: Journal Article,

Cardiac expression of 52beta, an alternative transcript of the congenital heart block-associated 52-kd SS-A/Ro autoantigen, is maximal during fetal development
Buyon JP; Tseng CE; Di Donato F; Rashbaum W; Morris A; Chan EK
1997 Apr;40(4):655-660, Arthritis & rheumatism
OBJECTIVE: Congenital heart block (CHB), associated with antibodies to SS-A/Ro and SS-B/La, is most often detected between 18 and 24 weeks of gestation, yet the maternal heart is unaffected. We recently described an alternatively spliced 52-kd SS-A/Ro messenger RNA (mRNA) derived from the skipping of exon 4 which encodes a smaller protein, 52beta (MW 45 kd), recognized by CHB maternal antisera. This study was designed to identify whether cardiac expression of 52beta and full-length 52alpha relates to the development of CHB. METHODS: Reverse transcriptase-polymerase chain reaction was performed using primers flanking exon 4 and mRNA from 22 human fetal hearts (age 11-25 weeks) and 3 adult hearts. The brain, kidney, liver, lung, and spleen were similarly evaluated in a 15-week, an 18-week, and a 24-week fetus. RESULTS: Expression of 52beta was greatest and 52alpha lowest between 14 and 16 weeks of gestation. In fetal hearts ages 22-25 weeks and adult heart, the 52beta transcript was markedly diminished and 52alpha clearly dominated. The 52beta mRNA was observed in a 15-week brain, kidney, lung, and spleen; however, its expression relative to 52alpha was greatest in the heart. CONCLUSION: Since expression of the alternative product 52beta is maximal at the time of cardiac ontogeny when maternal antibodies gain access to the fetal circulation, just prior to the clinical detection of bradyarrhythmia, a role for 52beta in the development of CHB is implicated. Although other fetal tissues express 52beta, there may be differences in accessibility of antigen or regenerative capacities
— id: 12333, year: 1997, vol: 40, page: 655, stat: Journal Article,

Recommendations for exogenous estrogen to prevent glucocorticoid-induced osteoporosis in premenopausal women with oligo- or amenorrhea: comment on the American College of Rheumatology recommendations for the prevention and treatment of glucocorticoid-induced osteoporosis
Buyon, J P; Dooley, M A; Meyer, W R; Petri, M; Licciardi, F
1997 Aug;40(8):1548-1549, Arthritis & rheumatism
— id: 73555, year: 1997, vol: 40, page: 1548, stat: Journal Article,

Connective tissue disease registries
Mayes, M D; Giannini, E H; Pachman, L M; Buyon, J P; Fleckman, P
1997 Sep;40(9):1556-1559, Arthritis & rheumatism
— id: 73553, year: 1997, vol: 40, page: 1556, stat: Journal Article,

Translocation of SSA/Ro and SSB/La in apoptotic human fetal cardiocytes
Miranda, E; Chan, EKL; Casiano, C; Tseng, C; DiDonato, F; Buyon, J
1997 SEP ;40(9):308-308, Arthritis & rheumatism
— id: 53182, year: 1997, vol: 40, page: 308, stat: Journal Article,

Induction of 52kD SSA/Ro antibodies and development of congenital heart block in a murine model
Miranda, E; Tseng, C; Chan, EKL; DiDonato, F; Boutidir, M; Buyon, J
1997 SEP ;40(9):553-553, Arthritis & rheumatism
— id: 53186, year: 1997, vol: 40, page: 553, stat: Journal Article,

Neonatal lupus syndromes
Tseng CE; Buyon JP
1997 Feb;23(1):31-54, Rheumatic diseases clinics of North America
Congenital heart block is considered to be a model of passively acquired autoimmunity, whereby immune abnormalities in the mother lead to the production of autoantibodies that cross the placenta and presumably injure the otherwise normally developing fetus. The major targets of the maternal immune response are the SSA/Ro and SSB/La ribonucleoproteins. Other neonatal abnormalities affecting the skin, liver, and blood elements have also been reported to be associated with anti-SSA/Ro-SSB/La antibodies in the maternal and fetal circulation and are now grouped along with congenital heart block under the heading of the neonatal lupus syndromes. This review covers the histopathology, SSA/Ro-SSB/La antigen-antibody systems, immunogenetics, clinical manifestations, and diagnosis and management strategies of these syndromes
— id: 12388, year: 1997, vol: 23, page: 31, stat: Journal Article,

The 52-kd protein as a target of intermolecular spreading of the immune response to components of the SS-A/Ro-SS-B/La complex
Tseng CE; Chan EK; Miranda E; Gross M; Di Donato F; Buyon JP
1997 May;40(5):936-944, Arthritis & rheumatism
OBJECTIVE: To determine whether immunization of healthy non-autoimmune mice with 52-kd SS-A/Ro induces a secondary antibody response to other components of the 48-kd SS-B/La-60-kd SS-A/Ro RNP complex and vice versa, since anti-52-kd antibodies have been invariably linked to these antigens in patients with Sjogren's syndrome and in mothers whose children have neonatal lupus. METHODS: Female BALB/c mice were immunized with 100 microg of 6xHis recombinant human 48-kd SS-B/La, 52-kd SS-A/Ro, or 60-kd SS-A/Ro proteins, or the 6xHis polypeptide control, each purified by Ni2+ affinity chromatography. Mice subsequently received booster injections with 50 microg of the same antigen every 10-21 days. Immune responses were measured by enzyme-linked immunosorbent assay (ELISA), immunoblotting of recombinant antigens, and immunoprecipitation of 35S-methionine-labeled in vitro translation products. RESULTS: Immunization with 48-kd SS-B/La resulted in anti-48-kd SS-B/La antibodies within 45 days, followed 10 days later by a secondary response to 52-kd SS-A/Ro, as measured by ELISA. Antibody spreading to 60-kd SS-A/Ro was not detected. Immunization with 52-kd SS-A/Ro resulted in rapid high-titer anti-52-kd SS-A/Ro responses within 27 days. Spreading to 48-kd SS-B/La occurred in only 1 mouse and 60-kd SS-A/Ro was detected in a minority of the mice after prolonged antigen exposure. Immunization with 60-kd SS-A/Ro led to anti-60-kd SS-A/Ro responses within 37 days, followed 3 months later by low-titer anti-48-kd SS-B/La and anti-52-kd SS-A/Ro antibodies. All primary immune responses were confirmed by immunoblotting and immunoprecipitation. While immunoblotting of the recombinant proteins revealed reciprocal intermolecular spreading in the majority of mice, immunoprecipitation clearly demonstrated that predominant spreading was generated after immunization with 48-kd SS-B/La, which consistently resulted in antibodies to 52-kd SS-A/Ro. CONCLUSION: The murine responses observed in the present study, demonstrating reciprocal intermolecular spreading to 48-kd SS-B/La, 52-kd SS-A/Ro, and 60-kd SS-A/Ro, support the linkage of 52-kd SS-A/Ro with the other proteins, despite their as-yet-undetected association in vivo. The marked recruitment of anti-52-kd SS-A/Ro responses elicited by 48-kd SS-B/La may provide a lead to exploring the physical interaction, direct or indirect, of 52-kd SS-A/Ro with the SS-A/Ro-SS-B/La RNP particle and its presentation to the immune system. These data should facilitate the establishment of a murine model of neonatal lupus
— id: 12320, year: 1997, vol: 40, page: 936, stat: Journal Article,

Arrhythmogenicity of IgG from mothers of children with congenital heart block
Boutjdir, M; Chen, L; Zhang, ZH; Tseng, C; DiDonato, F; ElSherif, N; Buyon, J
1996 OCT 15 ;94(8):4175-4175, Circulation
— id: 52748, year: 1996, vol: 94, page: 4175, stat: Journal Article,

IgG and affinity purified anti-52kD SSA/Ro antibodies from mothers of children with congenital heart block induce conduction defects and inhibit Ca channels in the human fetal heart
Boutjdir, M; Chen, L; Zhang, ZH; Tseng, C; DiDonato, F; Rashburn, W; Morris, W; ElSherif, N; Buyon, J
1996 OCT 15 ;94(8):3471-3471, Circulation
— id: 52747, year: 1996, vol: 94, page: 3471, stat: Journal Article,

Neonatal lupus
Buyon JP
1996 Sep;8(5):485-490, Current opinion in rheumatology
Neonatal lupus, albeit rare, affords an excellent opportunity to examine a disease from bedside to bench. Over the past year there have been numerous publications covering clinical aspects and basic research. The timing of heart block is not random; bradycardia is most often identified between 16 and 24 weeks of gestation. Investigations have focused on this apparently vulnerable period and examined the expression of known (SSA/Ro-SSB/La), novel (p57, endogenous retrovirus-3), and cross-reactive (laminin) autoantigens in fetal hearts of varied ages and in adult hearts. Clinical studies are accumulating, but a unique maternal autoantibody profile is yet to be identified. Anti-52-kD responses, measured by enzyme-linked immunosorbent assay and immunoblot, continue to be a nearly universal finding in mothers whose children have neonatal lupus. The presence of anti-U1RNP in the absence of anti-SSA/Ro-SSB/La antibodies occurs only in cases of isolated cutaneous disease and not (to date) in mothers of infants with cardiac manifestations. Immunogenetically, mothers with affected children appear to be more closely related to Sjogren's syndrome than systemic lupus erythematosus. Asymptomatic mothers do not invariably become ill, and if an asymptomatic mother does develop lupus it is not likely to be life threatening. Heart block is associated with substantial morbidity and mortality. Although treatment of affected fetuses with dexamethasone has successfully diminished associated effusions, this therapy has not reversed established third-degree block. Treatment with sympathomimetics may be beneficial in fetuses with hydropic changes. Prophylactic therapies, other than serial echocardiographic evaluation, are not supported by any published data. To further efforts at both the bench and bedside, research registries were recently established in the United States and Canada
— id: 12545, year: 1996, vol: 8, page: 485, stat: Journal Article,

The effects of pregnancy on autoimmune diseases
Buyon JP; Nelson JL; Lockshin MD
1996 Feb;78(2):99-104, Clinical immunology & immunopathology
Despite a now reasonable large body of developed information, critical gaps are apparent. Pregnancy is the only naturally occurring event in which an individual is exposed to nonself HLA. How the HLA mismatched fetus escapes rejection remains a biologic enigma. More definitive evidence is needed to determine if TH2 bias occurs in normal human pregnancy and how this impacts on established autoimmune diseases. With regard to specific autoantibody-associated placental and fetal disease, why aren't all pregnancies affected when the putative maternal antibodies are present? Future studies of maternal-fetal immunology, inclusive of information on cytokine regulation, should yield insights not only into the maintenance of normal pregnancy but also into the disproportionate increase of autoimmune diseases in females and the variable course of these diseases during pregnancy
— id: 12654, year: 1996, vol: 78, page: 99, stat: Journal Article,

Neonatal lupus: bedside to bench and back
Buyon, J P
1996 ;25(5):271-276, Scandinavian journal of rheumatology
Isolated congenital heart block (CHB) detected in utero is strongly associated with autoantibodies reactive with the intracellular soluble ribonucleoproteins 48 kD SSB/La, 52 kD SSA/Ro, and 60 kD SSA/Ro. An erythematous skin rash with a predilection for the scalp and periorbital area, most often apparent in the first few postnatal months, is also highly associated with these maternal antibodies. The permanent cardiac disease and transient cutaneous disease are the most common manifestations of the neonatal lupus syndromes. Fetal and neonatal disease are presumed to be due to the transplacental passage of these IgG autoantibodies from the mother, who may have systemic lupus erythematosus (SLE), Sjogren's syndrome, or be entirely asymptomatic, into the fetal circulation. The fetal heart appears to be uniquely vulnerable, since complete block has never been described in the mothers despite exposure to identical circulating levels of the autoantibodies. Extensive work from several laboratories has resulted in the molecular characterization of the maternal autoantibody responses and the cloning of genes expressing the cognate antigens whose structural features suggest a role in transcriptional regulation. While the precise pathogenetic mechanism of autoantibody mediated tissue injury is not defined it has been demonstrated in adult rabbit and human fetal hearts that sera containing anti-SSA/Ro antibodies induce atrioventricular block and inhibit L-type calcium currents (ICa) in isolated ventricular myocytes. From a clinical perspective the current data suggest that there is not a unique profile of maternal antibody response that predicts the development of neonatal lupus. However, a low risk profile is one in which the titer of anti-SSA/Ro antibodies is low and neither the 60 kD or 52 kD component is recognized on SDS-immunoblot and there are no detectable anti-SSB/La antibodies. Although the fetal disease is called neonatal lupus, this is clearly a misnomer since only about 25% of mothers actually fulfill criteria for the diagnosis of SLE. Furthermore, asymptomatic mothers do not invariably become ill and if an asymptomatic mother does develop SLE it is generally not life-threatening. The recurrence rare of CHB is low, about 15%, but this is nearly three times higher than the risk for CHB in a primigravida with the putative antibodies. CHB carries a significant mortality, 15-30%, and morbidity; two thirds of children require permanent pacing. While prospective clinical trials of fluorinated steroids in women with anti-SSA/Ro and/or anti-SSB/La antibodies is not likely to be initiated given the very low rate of CHB in these mothers, such trials in fetuses with heart block identified in utero are required before definitive recommendations can be made. Numerous anecdotal cases support the use of dexamethasone for treatment of effusions and hydrops and possibly incomplete block. To further efforts both at the bench and bedside, national registries have been established in the U.S. and Canada
— id: 73559, year: 1996, vol: 25, page: 271, stat: Journal Article,

Oral contraceptives in women with systemic lupus erythematosus
Buyon, J P
1996 ;147(4):259-264, Annales de medecine interne
Oral contraceptives (OCs) are generally not prescribed for women with SLE due to the widely-held view that they may activate disease. This practice is based on the greater incidence of SLE in women than in men, biologic abnormalities od estrogen metabolism, murine models of lupus, several anecdotes of patients having disease flares while receiving exogenous hormones, and a single retrospective study in patients with pre-existing renal disease. In contrast, recent retrospective patient surveys, albeit limited by the absence of formal analyses of disease activity, suggest that the rate of flare is not significantly increased in patients taking OCs. The preexisting data are insufficient to warrant the dismissal of a potentially important birth control option in a disease which predominantly affects women in their reproductive years and whose fertility is not altered by the disease. In counseling patients regarding lupus and pregnancy, there are clinical predictors of pregnancy outcome; patients in remission tend to do well. The same principles may be true regarding advice on the use of OCs; patients with inactive or stable/moderate disease and at low risk for thrombosis may benefit without a change in lupus activity. OCs with the lowest estrogen content should be used and consideration given to progestin only pills in situations where there is some risk of increased clotting. Large prospective double-blind placebo-controlled studies inclusive of all ethnic groups should provide the basis for more definitive recommendations
— id: 73558, year: 1996, vol: 147, page: 259, stat: Journal Article,

Identification of protein interactions of the Ro/La complex using the yeast two-hybrid system
Buyon, JP; DiDonato, F; Miranda, E; Chan, EKL
1996 SEP ;39(9):922-922, Arthritis & rheumatism
— id: 52782, year: 1996, vol: 39, page: 922, stat: Journal Article,

Identification and characterization of a novel mRNA transcript of the 60kD RO ribonucleoprotein encoding the N-terminal RNA binding domain only
Buyon, JP; DiDonato, F; Tseng, C; Rashbaum, W; Morris, A; Hamel, JC; Chan, EKL
1996 MAR ;44(3):A219-A219, Journal of investigative medicine
— id: 52939, year: 1996, vol: 44, page: A219, stat: Journal Article,

Congenital heart block: Mortality, morbidity, and recurrence rates obtained from a national neonatal lupus registry
Buyon, JP; Hiebert, R; Chin, D; Copel, J; Craft, J; Friedman, D; Lee, L; Provost, T; Reichlin, M; Rider, L; Rupel, A; Weston, W; Skovron, ML
1996 SEP ;39(9):1684-1684, Arthritis & rheumatism
— id: 52793, year: 1996, vol: 39, page: 1684, stat: Journal Article,

Increased fetal expression of a novel alternative 52RO mRNA transcript, 52 beta, coincides with vulnerability for congenital heart block (CHB)
Buyon, JP; Tseng, C; DiDonato, F; Rashbaum, W; Hamel, JC; Chan, EKL
1996 MAR ;44(3):A234-A234, Journal of investigative medicine
— id: 52946, year: 1996, vol: 44, page: A234, stat: Journal Article,

Cutaneous SLE: Relationship between serum Ro/La autoantibody profiles and Ro/La antigen expression in keratinocytes
Golden, BD; Tseng, CE; Belmont, HM; Buyon, JP
1996 SEP ;39(9):1582-1582, Arthritis & rheumatism
— id: 52792, year: 1996, vol: 39, page: 1582, stat: Journal Article,

On and off regulation of neutrophil adhesion accompanies phosphorylation of different beta(2) integrin residues
Merrill, JT; Guan, R; Shen, C; Buyon, JP
1996 MAR ;44(3):A239-A239, Journal of investigative medicine
— id: 52952, year: 1996, vol: 44, page: A239, stat: Journal Article,

Fine regulation of neutrophil adhesion: CR3 activation involves distinct phosphorylation changes induced by protein kinase C, serine-threonine and tyrosine phosphatases
Merrill, JT; Shen, C; Zhang, HW; Lahita, RG; Buyon, JP
1996 SEP ;39(9):61-61, Arthritis & rheumatism
— id: 52775, year: 1996, vol: 39, page: 61, stat: Journal Article,

Subclass distribution of maternal and neonatal anti-Ro(SSA) and La(SSB) antibodies in congenital heart block
Tseng CE; Caldwell K; Feit S; Chan EK; Buyon JP
1996 May;23(5):925-932, Journal of rheumatology
OBJECTIVE. To compare the subclass distribution of anti-48 kDa La(SSB) and anti-52 and 60 kDa Ro(SSA) antibodies in the maternal and neonatal circulation, in pregnancies affected and unaffected by the development of congenital heart block (CHB). METHODS. Sera were obtained from 32 mothers (during 34 pregnancies 23 complicated by CHB and 11 healthy) demonstrated to have anti-Ro(SSA) and/or La(SSB). Maternal and neonatal autoantibodies were evaluated for subclass distribution by ELISA. RESULTS. All 4 subclasses of anti-Ro(SSA) and La(SSB) antibodies cross the placenta and are detectable in sera obtained from the umbilical cord, IgG1 and IgG3 were the major subclasses represented in the 48 kDa La(SSB) and 52 kDa Ro(SSA) responses. All subclasses, including IgG2 and IgG4, were observed in about one-third of the anti-52 kDa Ro(SSA) and 48 kDa La(SSB) responses. In contrast, anti-60 kDa antibodies were, with rare exception, confined to IgG1. Except for anti-48 kDa La(SSB) IgG3 antibodies, no significant differences were observed between affected and unaffected pregnancies in the ratio of maternal to neonatal levels of any of the antibody subclasses. Overall, there were no significant differences in the subclass profiles between mothers whose children had heart block and those who did not. CONCLUSION. The IgG subclasses of anti-48 kDa La(SSB) and anti-52 and 60 kDa Ro(SSA) do not account for the susceptibility of one fetus versus another for the development of CHB. Anti-60 kDa Ro(SSA) antibodies are more restricted in subclass distribution than anti-52 kDA Ro(SSA) or 48 kDa La(SSB) responses
— id: 12614, year: 1996, vol: 23, page: 925, stat: Journal Article,

Stability of immunoblot profile of anti-SSA/Ro-SSB/La antibodies over time in mothers whose children have neonatal lupus
Tseng CE; Di Donato F; Buyon JP
1996 Jun;5(3):212-215, Lupus
Neonatal lupus is strongly associated with antibodies reactive with SSA/Ro-SSB/La proteins, independent of maternal disease activity or classification. We sought to determine whether the fine specificity of antibody profiles remains stable or evolves over time and whether these findings relate to clinical status. Sera from 23 mothers whose children had neonatal lupus (22 heart block, one skin) were evaluated by SDS-immunoblot. For each mother two samples were available at least 13 months apart; the mean duration of time between testing was 45 months +/- 27 S.D. (range 13-108 months). Twenty-two of the 23 initial profiles were identical to the results obtained in a later sample. The health status of seven (30%) of 23 mothers changed after the birth of the affected infant but the immunoblot specificity of the antibodies remained unchanged. SLE was the initial and final diagnosis in the only mother whose profiles differed, with development of weak reactivity to 48 kD SSB/La in addition to the 52kD SSA/Ro after 14 months. In conclusion, the fine specificity of anti-SSA/Ro-SSB/La antibodies as assessed by immunoblot is highly stable for years. Progression of clinical status was not associated with a concomitant change in antibody profile
— id: 12597, year: 1996, vol: 5, page: 212, stat: Journal Article,

The 52kD protein is a target of intermolecular spreading of the immune response to components of the Ro/La complex
Tseng, C; Chan, EKL; DiDonato, F; Buyon, JP
1996 SEP ;39(9):496-496, Arthritis & rheumatism
— id: 52780, year: 1996, vol: 39, page: 496, stat: Journal Article,

Discordant mRNA expression of 52 alpha, 52 beta, 48La, 60Ro autoantigens in cultured human cardiocytes and keratinocytes exposed to estradiol
Tseng, C; DiDonato, F; Wang, D; Chen, EKL; Buyon, JP
1996 SEP ;39(9):1131-1131, Arthritis & rheumatism
— id: 52784, year: 1996, vol: 39, page: 1131, stat: Journal Article,

Cloning and expression of mouse 60 kDa ribonucleoprotein SS-A/Ro
Wang, D; Buyon, J P; Chan, E K
1996 ;23(3-4):205-210, Molecular biology reports
Autoantibodies to SS-A/Ro are among the most common found in sera of patients with systemic rheumatic diseases. These autoimmune diseases can affect various organ systems of the body and are variable in their manifestations and presentation. One of the autoimmune targets is the 60 kDa SS-A/Ro protein known to be associated with small cytoplasmic Y RNAs. To study systematically the expression of the protein, we have cloned the mouse full length 60 kDa SS-A/Ro cDNA using 5' RACE based on a cDNA sequence reported in the mouse genome project. The recombinant protein derived from the putative full-length construct was shown to react with human prototype anti-SS-A/Ro serum Ge in western blot and immunoprecipitation and comigrated with cellular 60 kDa SS-A/Ro protein in 3T3 cells. Cellular expression, measured by RT-PCR, was highest in mouse brain, followed by lung, muscle, kidney and heart. Lower levels were found in testis, liver and spleen. Like the human 60 kDa SS-A/Ro protein, the deduced mouse homolog has 538 amino acids. Sequence analysis showed 89.9% identity and 95.0% similarity between the mouse and human proteins
— id: 73557, year: 1996, vol: 23, page: 205, stat: Journal Article,

Misoprostol and Prednisone Treatment of Lupus Nephritis
Belmont HM; Kitsis E; Skovron ML; Buyon J; McCullagh E; Abramson S
1995 Dec;2(12):928-932, American journal of therapeutics
Fourteen patients were enrolled in a placebo-controlled double-blind randomized trial of 8 weeks of treatment for active lupus nephritis. Seven patients received prednisone at a dose of 1 mg kg(minus sign1) day(minus sign1) plus misoprostol at a dose of 200 &mgr;g P.O. Q.I.D.; 7 patients received prednisone plus placebo. The patients included 12 females, 2 males; 3 African-Americans, 3 Asians, 5 Hispanics, and 3 Caucasians. There were no serious side effects associated with prednisone or misoprostol treatment during the 8-week study period. Laboratory measures obtained at baseline, 4, 8, and 12 weeks included complete blood count (CBC), ESR, C reactive protein (CRP), serum creatinine, creatinine clearance, 24-h urine protein excretion, C3, C4, and anti-double stranded DNA (anti-dsDNA). Statistical analysis was conducted assessing change in measures over time in the entire study group by paired t-tests. The effect of treatment on change over time was examined by analysis of covariance. Log transformation of the variables was performed prior to statistical analysis. For the entire study group, the mean levels of ESR, CRP, 24-h protein excretion, C3, C4, and anti-dsDNA were improved at 4, 8, and 12 weeks. The mean ESR at baseline was 70 plus minus 8 compared to 42 plus minus 8 at 12 weeks (p < 0.01). The mean CRP at baseline was 0.6 plus minus 0.2 compared to 0.2 plus minus 0.1 at 12 weeks (p < 0.01). The 24-h protein excretion was 4367 plus minus 769 mg at baseline compared to 2512 plus minus 709 mg at 12 weeks (p = 0.02). The mean C3 at baseline was 40 plus minus 4 mg dl(minus sign1) compared to 60 plus minus 4 mg dl(minus sign1) at 12 weeks (p < 0.01). The mean C4 at baseline was 14 plus minus 1 mg dl(minus sign1) compared to 23 plus minus 2 mg dl(minus sign1) at 12 weeks (p < 0.01). The mean anti-dsDNA at baseline was 4268 plus minus 1780 compared to 316 plus minus 111 at 12 weeks (p < 0.001). The baseline serum creatinine (1.12 plus minus.15 mg dl(minus sign1)) and creatinine clearance (82 plus minus 15 ml min(minus sign1)) were not significantly changed at 12 weeks (1.10 plus minus 0.2 mg dl(minus sign1) and 80 plus minus 17 ml min(minus sign1), respectively). Comparing the misoprostol treatment group to the placebo group, there were no statistically significant differences for ESR, CRP, creatinine, creatinine clearance, 24-h protein excretion, C3, C4, or anti-dsDNA at any time points. However, comparing the misoprostol treatment group at 4 weeks to the placebo group, a more rapid decrease in anti-dsDNA (reduction of 3297 plus minus 2374) was observed in the misoprostol treatment group versus 304 plus minus 409 in the placebo group), as well as lower mean anti-dsDNA levels (464 plus minus 140) in the misoprostol treatment group versus 4118 plus minus 3834 in the placebo group). Also, the C3 and C4 levels at 12 weeks in the misoprostol treatment group (67 plus minus 5 and 27 plus minus 3 mg dl(minus sign1), respectively) were greater than levels in the placebo group (52 plus minus 4 and 19 plus minus 3 mg dl(minus sign1), respectively). The data demonstrate that oral prednisone is effective in reducing proteinuria and improving the biological markers of disease activity (i.e., ESR, CRP, C3, C4, and anti-dsDNA) following short-term treatment of active lupus nephritis. Additionally, the more rapid change in anti-dsDNA levels and superior normalization of complement levels in the treatment group, although not statistically significant, are consistent with a biologic effect of misoprostol on lymphocyte function and the production of a pathogenic autoantibodies
— id: 39703, year: 1995, vol: 2, page: 928, stat: Journal Article,

New York University/Hospital for Joint Diseases experience with intravenous cyclophosphamide treatment: efficacy in steroid unresponsive lupus nephritis
Belmont HM; Storch M; Buyon J; Abramson S
1995 Apr;4(2):104-108, Lupus
The medical records of patients receiving cyclophosphamide for lupus nephritis between 1987 and 1993 at the New York University/Hospital for Joint Diseases Lupus Study Group Institutions were retrospectively reviewed. We identified 45 patients (38 female, seven male) who received a mean of 9 +/- 1 (range 2-23) pulses of intravenous cyclophosphamide for diffuse proliferative glomerulonephritis (n = 28), focal proliferative glomerulonephritis (n = 7), membranous nephropathy (n = 5), mesangial nephropathy with sclerosis (n = 1) or nephritis without biopsy (n = 4). Forty-two of the 45 patients received cyclophosphamide after failing steroid therapy. During a follow-up period of 52 +/- 3 months, nine patients progressed to end-stage renal disease (ESRD) with three additional patients experiencing a doubling of the creatinine and two patients persistent nephrotic range proteinuria. There were no deaths directly attributable to cyclophosphamide and no patients developed hemorrhagic cystitis or malignancy. Ten of 37 women had ceased menstruating prior to cyclophosphamide therapy. Treatment-associated amenorrhea occurred in only three patients all over 27 years of age. Intermittent intravenous cyclophosphamide therapy of lupus nephritis is well tolerated and usually effective in maintaining renal function in patients unresponsive to steroids although, in our experience, 20% of patients developed ESRD and a total of 14 of 45 (30%) patients had unsatisfactory outcomes
— id: 57339, year: 1995, vol: 4, page: 104, stat: Journal Article,

PROSPECTIVE-STUDY OF PREDNISONE TREATMENT IN ACTIVE LUPUS NEPHRITIS - EXPERIENCE WITH STEROID AS A FIRST LINE AGENT
BELMONT, HM; BUYON, J; SKOVRON, ML; MCCULLAGH, E; KITSIS, E; ABRAMSON, SB
1995 SEP ;38(9):909-909, Arthritis & rheumatism
— id: 86697, year: 1995, vol: 38, page: 909, stat: Journal Article,

UP-REGULATED EXPRESSION OF INDUCIBLE NITRIC-OXIDE SYNTHASE IN SLE - EVIDENCE FOR ACTIVATED ENDOTHELIUM
BELMONT, HM; LEVARTOVSKY, D; AMIN, AR; SKOVRON, ML; BUYON, J; GIORNO, R; REDISKE, J; ABRAMSON, SB
1995 SEP ;38(9):1422-1422, Arthritis & rheumatism
— id: 86701, year: 1995, vol: 38, page: 1422, stat: Journal Article,

ELECTROPHYSIOLOGIC CHARACTERIZATION OF PURIFIED IGG FROM A MOTHER WHOSE CHILD HAS CONGENITAL HEART-BLOCK (CHB) ON L-TYPE CALCIUM CURRENTS (I-CA)
BOUTJDIR, M; ZHANG, ZH; CHEN, L; ELSHERIF, N; TSENG, CE; DIDONATO, F; RASHBAUM, W; BUYON, JP
1995 SEP ;38(9):466-466, Arthritis & rheumatism
— id: 86695, year: 1995, vol: 38, page: 466, stat: Journal Article,

Can Women with Systemic Lupus Erythematosus Safely Use Exogenous Estrogens?
Buyon JP; Kalunian KC; Skovron ML; Petri M; Lahita R; Merrill J; Sammaritano L; Yung C; Licciardi F; Belmont HM; Hahn BH
1995 Aug;1(4):205-212, Journal of clinical rheumatology. JCR
The current study was initiated to estimate the use of oral contraceptives and estrogen replacement therapy in women with systemic lupus erythematosus (SLE). Four hundred and four patients were surveyed from five medical centers. Two hundred and twenty four (55%) had ever used oral contraceptives, however, only 51 (13%) were taking oral contraceptives at the time SLE was diagnosed. Fifty five (14%) used oral contraceptives after their disease was diagnosed. Only seven (13%) reported an exacerbation of disease activity, mostly confined to the musculoskeletal system. In one substudy, there were no significant differences observed between women with or without SLE with regard to the frequency of ever-use of oral contraceptives. In contrast, significantly fewer women with established SLE were taking oral contraceptives at the time of interview compared with healthy women, p < 0.02. In a second substudy, information on past and present usage of estrogen replacement therapy was obtained in women followed at two of the sites included in the main study. Fifty-five (59%) of the 94 postmenopausal patients at these centers had ever taken estrogen therapy, 23 (24%) at the time of diagnosis. Forty-eight women (51%) began or remained on estrogen therapy after the diagnosis of SLE, four (8%) of whom reported exacerbations of disease activity. A significantly higher percentage of Caucasian women had taken or were taking estrogen replacement compared with other ethnic groups. This study suggests that exogenous hormones were generally well tolerated by women with SLE; this preliminary observation is based on patient recall. The low frequency of current oral contraceptive use in lupus patients of reproductive age may reflect, in part, bias of the managing rheumatologists and obstetricians/gynecologists. Given the health needs of and potential benefits for women with SLE, these observations suggest that larger prospective studies are critical and are likely to change prescribing practices for exogenous estrogen
— id: 114627, year: 1995, vol: 1, page: 205, stat: Journal Article,

In utero identification and therapy of congenital heart block
Buyon JP; Waltuck J; Kleinman C; Copel J
1995 Apr;4(2):116-121, Lupus
To determine the feasibility and effectiveness of prenatal therapy of congenital heart block (CHB), information was sought regarding the timing of identification of CHB in 72 pregnancies and the outcome of those pregnancies in which the mothers were administered corticosteroids. Mailed questionnaires with telephone follow-up, data from primary physicians and chart review were utilized. In 38 (53%) of affected pregnancies CHB was identified between 16 and 24 weeks of gestation, in 17 (24%) between 25 and 30 weeks of gestation, in 8 (11%) between 31 and 37 weeks, and in 5 (7%) between 38 and 40 weeks. In four pregnancies the timing was unknown. Five women were taking prednisone for disease activity prior to the discovery of CHB and in six other women prednisone therapy was initiated at or about the time CHB was identified. In 19 pregnancies, women were given fluorinated steroids (available to the fetus in an active form) as attempted therapy after the discovery of CHB. Of these, one fetus with second degree block reverted to sinus rhythm and two with third degree block exhibited an improvement in the degree of block. In eight fetuses pleural and/or pericardial effusions resolved. In conclusion, the gestational period of heightened fetal vulnerability for the development of heart block is in the mid second to early third trimester. Although there is no evidence that maternal prednisone should be used prophylactially, fluorinated steroids may be efficacious after the identification of heart block, particularly with regard to an associated myocarditis
— id: 56617, year: 1995, vol: 4, page: 116, stat: Journal Article,

Postmenopausal hormone therapy and systemic lupus erythematosus
Buyon, J P; Belmont, H M; Kalunian, K C
1995 Dec 15;123(12):961-961, Annals of internal medicine
— id: 73560, year: 1995, vol: 123, page: 961, stat: Journal Article,

DIFFERENTIAL EXPRESSION OF AN ALTERNATIVELY SPLICED TRANSCRI
BUYON, JP; DIDONATO, F; TSENG, CE; HAMEL, JC; CHAN, EKL
1995 SEP ;38(9):919-919, Arthritis & rheumatism
— id: 86698, year: 1995, vol: 38, page: 919, stat: Journal Article,

60KD RO - IDENTIFICATION OF A NOVEL MESSENGER-RNA TRANSCRI
BUYON, JP; DIDONATO, F; TSENG, CE; RASHBAUM, W; HAMEL, JC; CHAN, KL
1995 SEP ;38(9):351-351, Arthritis & rheumatism
— id: 86691, year: 1995, vol: 38, page: 351, stat: Journal Article,

52-kD SS-A/Ro: genomic structure and identification of an alternatively spliced transcript encoding a novel leucine zipper-minus autoantigen expressed in fetal and adult heart
Chan EK; Di Donato F; Hamel JC; Tseng CE; Buyon JP
1995 Oct 1;182(4):983-992, Journal of experimental medicine
The 52-kD SS-A/Ro protein is one of the antigenic targets strongly associated with the autoimmune response in mothers whose children have manifestations of neonatal lupus. In addition to the cDNA clone we previously reported for the full-length 52-kD SS-A/Ro protein, an interesting MOLT-4 cDNA clone, p52-2, was found to have an internal deletion of 231 nucleotides including the domain encoding the leucine zipper motif. To further investigate the nature of this deletion, genomic DNA clones were isolated from a lambda FIXII library. The complete gene for the full-length 52-kD protein (alpha form, 52 alpha) spans 10 kb of DNA and is composed of seven exons. Exon 1 contains only the 5' untranslated sequence, while the translation initiation codon is located 3 kb downstream in exon 2, which also encodes the three zinc finger motifs. Exon 4 encodes amino acids 168-245, including the coiled coil/leucine zipper domain. Exon 7 is the longest and encodes the rfp-like domain and the 3' untranslated region. The cDNA p52-2 can now be accounted for as a product of alternative messenger RNA (mRNA) derived from the splicing of exon 3 to exon 5, skipping exon 4, which results in a smaller protein (52 beta) with a predicted molecular weight of 45,000. An initial approach to identifying this alternatively spliced form in the human heart used a ribonuclease protection assay. Using an RNA probe corresponding to bases 674-964 of the full-length cDNA, two protected mRNA fragments were identified, a 290-bp fragment corresponding to expression of 52 alpha and a smaller fragment of 144 bp, the predicted size of 52 beta. Using reverse transcription followed by polymerase chain reaction, cDNAs from a 16-wk fetal heart, 24-wk heart, and adult heart were amplified with primers flanking exon 4. Two polymerase chain reaction products were observed in each tissue, one 1.0 kb likely representing 52 alpha and a second 0.78 kb, consistent with 52 beta. The 0.78-kb fragment identified in the 16-wk heart was cloned, and DNA sequencing confirmed the 52 beta type. Immunoprecipitation of in vitro-translated 35S-labeled 52 beta form was performed to evaluate the antigenicity of this novel form of 52-kD SS-A/Ro. 26 (87%) of 30 sera tested from mothers whose children were known to have neonatal lupus immunoprecipitated the 52 beta form.(ABSTRACT TRUNCATED AT 400 WORDS)
— id: 18582, year: 1995, vol: 182, page: 983, stat: Journal Article,

Successful in utero therapy of fetal heart block
Copel JA; Buyon JP; Kleinman CS
1995 Nov;173(5):1384-1390, American journal of obstetrics & gynecology
OBJECTIVE: Congenital complete heart block is associated with maternal autoantibodies to Ro and La proteins, which injure the fetal cardiac conduction system. We administered dexamethasone to the mothers of five fetuses with heart block caused by maternal antibodies. STUDY DESIGN: We diagnosed five cases of fetal heart block at 20 to 23 weeks and treated all mothers with dexamethasone 4 mg orally each day for the remainder of the pregnancy. All patients were positive for anti-SS-A (anti-Ro) and/or anti-SS-B (anti-La) antibodies. RESULTS: Four patients had complete heart block, and one had second-degree block. In two patients (one with complete heart block, one with second-degree heart block) the degree of block lessened with treatment. Hydrops in three complete heart block patients resolved after treatment. Maternal antibody levels did not change. Matched maternal and cord samples at delivery showed similar antibody levels. CONCLUSIONS: Complete heart block can respond to transplacental glucocorticoid therapy with improved cardiac conduction. Because there may be a concurrent myocarditis, treatment in utero may also improve cardiac contractility, leading to the observed rapid resolution of hydrops. Treatment with steroids that cross the placenta should be considered for newly diagnosed cases of complete heart block with positive antibody screens
— id: 62316, year: 1995, vol: 173, page: 1384, stat: Journal Article,

Benign fetal bradycardias diagnosed by echocardiography
Friedman, D M; Borg, M; Rutkovsky, L; Buyon, J P
1995 Mar;12(2):87-90, American journal of perinatology
Five cases of benign fetal arrhythmias presented with dramatic pictures of sustained irregular bradycardias. These cases were not associated with structural heart disease, the development of hydrops fetalis, or maternal autoantibodies. In all but one case, the arrhythmias resolved before delivery. Two fetuses had 2:1 atrioventricular block and the other three showed group beating due to blocked atrial trigeminy. With regular intermittent reevaluation, and if no signs of hydrops occur, watchful waiting is the suggested therapeutic modality
— id: 73561, year: 1995, vol: 12, page: 87, stat: Journal Article,

Ontogeny of membrane cofactor protein: phenotypic divergence in the fetal heart
Gorelick A; Oglesby T; Rashbaum W; Atkinson J; Buyon JP
1995 Aug;4(4):293-296, Lupus
Human adult cells are protected from complement-induced damage in part by membrane cofactor protein (MCP, CD46). To examine fetal characteristics which might influence autoantibody-mediated diseases acquired in utero, such as heart block in neonatal lupus, the tissue expression of MCP was studied. Using a high ratio of acrylamide:bisacrylamide, immunoblots of tissues from six fetuses (aged 19-24 weeks) probed with rabbit anti-MCP antibodies revealed a band at 60 KD in addition to the known 65 KD and 55 KD isoforms which comprise the codominant allelic system of MCP. Five fetuses expressed the most common MCP polymorphism (predominance of the 65 KD isoform, upper band alpha-phenotype) in the kidney, spleen, liver and lung. In contrast, all hearts from these five fetuses demonstrated a different pattern in which there was a marked decrease in the intensity of the 65 KD band and accentuation of the lower molecular weight bands. In a sixth fetus, which expressed the second most common polymorphism (equal expression of the 65 KD and 55 KD MCP isoforms, alpha beta-phenotype), the heart was similar to the other tissues. These studies confirm the expression of MCP in early gestational life. Preferential expression of the MCP beta-isoform in the majority of fetal hearts irrespective of the phenotype of other organs, suggests tissue-specific RNA splicing or post-translational modification which may relate to autoantibody-mediated injury in diseases such as neonatal lupus
— id: 56757, year: 1995, vol: 4, page: 293, stat: Journal Article,

Heterogeneity in the expression of Ro and La antigens in human skin
Niimi Y; Ioannides D; Buyon J; Bystryn JC
1995 Sep;38(9):1271-1276, Arthritis & rheumatism
OBJECTIVE. To determine whether there are variations in the expression of Ro and La antigens in human skin. METHODS. Levels of expression of Ro and/or La antigens in 26 specimens of normal human skin (11 sun-exposed, 15 sun-protected) were measured by indirect immunofluorescence with monospecific antisera. RESULTS. Levels of expression of both antigens varied by more than 2,000-fold in the skin of different individuals. There usually was a correlation between the levels of expression of Ro and La antigens in the same skin specimen. There was no correlation found between the levels of Ro or La antigen expression and sun exposure, nor was there a correlation found between levels of antigen expression and location of the skin on the body. CONCLUSION. There is a marked heterogeneity in the expression of both Ro and La antigens in the skin of different individuals. The present study findings suggest that the levels of expression of these antigens may play a role in the propensity of some individuals to develop anti-Ro or anti-La antibodies and/or skin lesions associated with the presence of these antibodies
— id: 56784, year: 1995, vol: 38, page: 1271, stat: Journal Article,

AUTOANTIBODY RESPONSE TO THE RO/LA PARTICLE MAY PREDICT OUTCOME IN NEONATAL LUPUS-ERYTHEMATOSUS
SILVERMAN, ED; BUYON, J; LAXER, RM; HAMILTON, R; BINI, P; CHU, JL; ELKON, KB
1995 JUN ;100(3):499-505, Clinical & experimental immunology
This study was undertaken to determine the role of antibodies against both recombinant Ro (r-Ro) and La (r-La) proteins and polypeptides derived from the recombinant La protein in predicting fetal and neonatal outcome in children at risk to develop neonatal lupus erythematosus (NLE). All sera were obtained in the perinatal period and quantitative ELISA assays were used. We collected 41 maternal sera within 2 months of delivery of a child with NLE (21 with congenital heart disease block (CHB) and 20 with dermatologic NLE) and 19 sera from anti-Re and/or anti-La antibody-positive mothers with systemic lupus erythematosus (SLE) who delivered a child without NLE. All sera were tested for anti-r-La and anti-r-Ro antibodies by ELISA, and most sera were tested for antibodies directed against La polypeptides by immunoblot. We found significantly higher anti-r-La La antibody levels in the sera from mothers of children with NLE compared with sera from mothers of unaffected children (0.67 +/- 0.43 versus 0.14 +/- 0.30; P < 0.0001). There was a statistically significant difference in the mean anti-r-La levels between the sera of mothers of children with CHB compared with dermatologic NLE (0.51 +/- 0.45 versus 0.83 +/- 0.37 respectively; P = 0.0091). When we examined antibodies directed against the recombinant 52-kD Ro protein, there was a statistically significant elevation of titres in the sera of mothers of NLE children (0.77 +/- 0.35) compared with non-NLE mothers (0.29 +/- 0.39; P < 0.0001). There was no difference in the r-Ro levels between mothers of children with dermatologic NLE compared with CHB (0.82 +/- 0.37 versus 0.71 +/- 0.74; P = 0.32). When we examined polypeptides derived from the recombinant La protein, the mean number of polypeptides recognized by sera from mothers of children with NLE was significantly higher than the mean number of polypeptides recognized by sera from mothers of unaffected children (5.1 +/- 0.54 versus 2.3 +/- 0.54 respectively; P < 0.001). More importantly, when we examined the individual polypeptides, we found that only sera from mothers of children with NLE and not from mothers of unaffected children recognized a polypeptide designated DD (30% versus 0%, respectively). These studies indicate that the autoantibody response to the Ro/La particle can differentiate sera from mothers of children with NLE and sera from mothers of unaffected children. Furthermore, there was a difference in the anti-La autoantibody response between mothers of children with CHB and dermatologic NLE. Antibodies directed against the DD polypeptide derived from the recombinant La protein were seen only in the sera of mothers of children with NLE
— id: 87288, year: 1995, vol: 100, page: 499, stat: Journal Article,

RELATIONSHIP OF FINE SPECIFICITY OF ANTI-RO/LA ANTIBODIES TO CLINICAL STATUS OVER TIME IN MOTHERS OF CHILDREN WITH NEONATAL LUPUS
TSENG, CE; GOLDEN, B; BUYON, JP
1995 SEP ;38(9):396-396, Arthritis & rheumatism
— id: 86692, year: 1995, vol: 38, page: 396, stat: Journal Article,

Up-regulation of endothelial cell adhesion molecules characterizes disease activity in systemic lupus erythematosus. The Shwartzman phenomenon revisited
Belmont HM; Buyon J; Giorno R; Abramson S
1994 Mar;37(3):376-383, Arthritis & rheumatism
OBJECTIVE. To test the hypothesis that during exacerbations of systemic lupus erythematosus (SLE), endothelial cells are activated to increase their expression of adhesion molecules. METHODS. Endothelial cell expression of E-selectin, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule 1 (ICAM-1) was quantitated immunohistochemically in 20 biopsy specimens from nonlesional, non-sun-exposed skin from 16 SLE patients. Disease activity was evaluated with the SLE Disease Activity Index (SLEDAI) and with measurements of complement components C3a desArg, C3, and C4. RESULTS. The mean expression of all 3 adhesion molecules was significantly elevated in patients with SLE versus healthy controls, as well as in patients with active versus inactive SLE. The mean C3a desArg level was significantly higher in patients with active SLE compared with those with inactive SLE. The SLEDAI scores correlated directly with C3a desArg levels and inversely with C3 and with C4 levels. Evaluation of serial biopsy specimens demonstrated loss of endothelial cell adhesion molecules and reduction of C3a levels with clinical improvement. CONCLUSION. Our findings demonstrate up-regulation of the surface expression of 3 distinct adhesion molecules, E-selectin, VCAM-1, and ICAM-1, in patients with SLE. The abnormal expression of these endothelial cell adhesion molecules is most marked in patients with active disease characterized by significant elevations of the complement split product C3a desArg. We suggest that in certain SLE patients, excessive complement activation in association with primed endothelial cells induces leukocyte-endothelial cell adhesion and leuko-occlusive vasculopathy
— id: 56498, year: 1994, vol: 37, page: 376, stat: Journal Article,

Autoantibody responses to the "native" 52-kDa SS-A/Ro protein in neonatal lupus syndromes, systemic lupus erythematosus, and Sjogren's syndrome
Buyon JP; Slade SG; Reveille JD; Hamel JC; Chan EK
1994 Apr 1;152(7):3675-3684, Journal of immunology
Abs to the 52-kDa SS-A/Ro protein are found in high prevalence in patients with Sjogren's syndrome (SS) and mothers whose children have the neonatal lupus syndrome (NLS). This study further defines the specificity of this response. By ELISA, 97% of 59 mothers of offspring with NLS had Abs to the 52-kDa recombinant protein compared with 80% in 132 non-NLS sera with anti-SS-A/Ro Abs (p 0.004). Antigenic regions on the 52-kDa protein were evaluated by immunoprecipitation of [35S]-radiolabeled in vitro translation products. Ninety-five percent of 99 sera that contained anti-52-kDa Abs by ELISA reacted with a large fragment spanning amino acids (aa) 1-291. Two antigenic regions were identified, aa169-291 containing the leucine zipper that was recognized by 83% of the anti-52-kDa sera tested and aa1-78 containing the zinc finger domains that was recognized by only half the sera. No sera immunoprecipitated this N-terminal fragment exclusively. Recognition of one or both regions was not unique to any clinical subset of patients; however, a greater number of sera from patients with SS contained both specificities, whereas asymptomatic mothers whose children had NLS comprised the only clinical group in which the majority recognized the central region of the molecule. Reactivity with both epitopes was demonstrated significantly more often in sera with high titers of Abs to the 60-kDa rSS-A/Ro protein by ELISA in association with the anti-52-kDa response compared with anti-52-kDa responses associated with low titers of anti-60-kDa Abs (p < 0.04). Eighty-one percent of 16 sera that recognized the N-terminal epitope were from patients with the combination of HLA-DRB1*0301, DQA1*0501, and DQB1*0201 alleles, compared with 30% of 10 that recognized only the central epitope (p < 0.02). In summary, this study demonstrates that there are at least two antigenic determinants on the 52-kDa SS-A/Ro protein, one 'immunodominant' and the other recognized by a more 'restricted' subset of anti-52-kDa SS-A/Ro Abs
— id: 12988, year: 1994, vol: 152, page: 3675, stat: Journal Article,

Relationship between maternal and neonatal levels of antibodies to 48 kDa SSB(La), 52 kDa SSA(Ro), and 60 kDa SSA(Ro) in pregnancies complicated by congenital heart block
Buyon JP; Waltuck J; Caldwell K; Crawford B; Slade SG; Copel J; Chan EK
1994 Oct;21(10):1943-1950, Journal of rheumatology
OBJECTIVE. Since there is suggestive but, to date, indirect evidence that maternal anti-SSA(Ro) and SSB(La) antibodies are pathogenic in congenital heart block (CHB), we explored the hypothesis that binding to fetal tissues would result in selective depletion of autoantibodies from the neonatal circulation. METHODS. Maternal and umbilical cord levels of anti-48 kDa SSB(La), anti-52 kDa SSA(Ro) and anti-60 kDa SSA(Ro) antibodies were measured by ELISA and immunoprecipitation at parturition in 15 pregnancies complicated by CHB. A control group consisted of 13 pregnancies in which the mother was known to have antibodies to either SSA(Ro) and/or SSB(La) and the children did not have CHB. RESULTS. The ratios of maternal to cord serum levels of anti-48 SSB(La), anti-52 SSA(Ro) and anti-60 kDa SSA(Ro) antibodies ranged from 0.71 to 2.38 in both affected and unaffected pregnancies. The mean ratio obtained for each of the 3 autoantibodies was not significantly different between the 2 groups. Moreover these ratios did not significantly differ from the mean ratios obtained for total IgG levels in either group. CONCLUSION. These data demonstrate that maternal antibodies to all components of the SSA(Ro) SSB(La) system are efficiently transported across the placenta and are not selectively depleted in the circulation of neonates with CHB
— id: 7894, year: 1994, vol: 21, page: 1943, stat: Journal Article,

Neonatal lupus syndromes
Buyon, J P
1994 Sep;6(5):523-529, Current opinion in rheumatology
Neonatal lupus continues to generate considerable interest despite its rarity; more than 15 original contributions were made to the literature in the past year. Diverse aspects of this 'syndrome' of passively acquired autoimmunity have been covered. Experiments using a rabbit model provided insights into the pathogenicity of maternal anti-Ro/SS-A and anti-La/SS-B antibodies. Perfusion of rabbit hearts with anti-Ro/SS-A and anti-La/SS-B sera resulted in conduction abnormalities in whole adult rabbit hearts and induced a reduction in the peak slow inward current in patch-clamp experiments of isolated rabbit ventricular myocytes, suggesting involvement of calcium channels. Clinical investigations are moving away from case reports, and recent studies now include substantial entries. Assuming that patients reported from the United States, Finland, and England are all separate, sera from at least 100 different mothers of infants with congenital heart block have been studied. Although there is apparently no serologic profile that is unique to mothers of affected children, compared with mothers of healthy children, anti-Ro/SS-A antibodies (anti-52-kD antibodies are more prevalent by immunoblot in congenital heart block, although all these sera are likely to have anti-60-kD antibodies by immunoprecipitation) are usually of high titer and associated with anti-La/SS-B antibodies. To date, the only maternal autoantibodies that have been associated with congenital heart block recognize Ro/SS-A or La/SS-B antigens. Mothers of affected infants are often asymptomatic, and when symptomatic, the clinical features are frequently characteristic of Sjogren's syndrome. Although treatment of affected fetuses with dexamethasone has successfully diminished associated effusions, there has been no report of reversal of established third-degree heart block
— id: 73562, year: 1994, vol: 6, page: 523, stat: Journal Article,

ONTOGENY OF COMPLEMENT REGULATORY PROTEINS - DISCORDANT EXPRESSION IN THE FETAL HEART
GORELICK, A; OGLESBY, T; RASHBAUM, W; ATKINSON, J; BUYON, JP
1994 APR ;42(2):A272-A272, Clinical research
— id: 52497, year: 1994, vol: 42, page: A272, stat: Journal Article,

Dynamic state of beta 2 integrin phosphorylation: regulation of neutrophil aggregation involves a phosphatase-dependent pathway
Merrill, J T; Winchester, R J; Buyon, J P
1994 May;71(2):216-222, Clinical immunology & immunopathology
The role of phosphorylation and dephosphorylation events in homotypic neutrophil aggregation mediated by CD11b/CD18 molecules was investigated using okadaic acid, an inhibitor of serine and threonine phosphatases. In the absence of exogenous stimuli the addition of okadaic acid to neutrophils resulted in a dose-dependent increase in phosphorylation of the CD18 beta chain that was further augmented by PMA but unaffected by FMLP. Phosphorylation induced by okadaic acid was reversed by staurosporine and minimally decreased by the less selective PKA/PKC inhibitors, H-7 and H-8. This suggests the existence of constitutive phosphatase and kinase activity emphasizing the dynamic state of phosphorylation and dephosphorylation of the beta 2 integrins. Unlike PMA, okadaic acid did not promote homotypic neutrophil aggregation. Furthermore, both the PMA-induced pathway of irreversible aggregation, blocked by staurosporine, as well as the FMLP-induced pathway of reversible aggregation, augmented by staurosporine, were inhibited by okadaic acid in a dose- and time-dependent manner. These results provide evidence that a phosphatase-dependent step is involved in each of these two distinct pathways that regulate neutrophil aggregation mediated by beta 2 integrin activation
— id: 73563, year: 1994, vol: 71, page: 216, stat: Journal Article,

Intravascular neutrophil activation in systemic lupus erythematosus (SLE): dissociation between increased expression of CD11b/CD18 and diminished expression of L-selectin on neutrophils from patients with active SLE
Molad Y; Buyon J; Anderson DC; Abramson SB; Cronstein BN
1994 Jun;71(3):281-286, Clinical immunology & immunopathology
Previous studies have shown that neutrophils in the circulation of patients with active systemic lupus erythematosus (SLE) are activated as judged by their increased surface expression of the beta 2-integrin CD11b/CD18. Since activation of neutrophils leads to altered expression of another adhesion molecule, L-selectin (LS), we examined neutrophils from patients with SLE for changes in the expression of CD11b/CD18 and LS by cytofluorographic analysis of immunofluorescent-labeled cells. Overall there was no difference between surface expression of CD11b/CD18 on neutrophils from SLE patients or controls [mean fluorescence 225 +/- 26 vs 225 +/- 13 relative fluorescence units (RFU), respectively]. However, as previously reported, neutrophils from patients with more active disease (activity score > or = 3, UCH Middlesex activity score) expressed greater CD11b/CD18 than neutrophils from controls (319 +/- 40 RFU, P < 0.03, n = 9) or from patients with less active disease (193 +/- 10 RFU, P < 0.006). Indeed, CD11b/CD18 expression correlated directly with disease activity (r = 0.54, P < 0.02). Stimulation of neutrophils ex vivo with the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (100 nM) induced up-regulation of CD11b/CD18 in cells from both SLE patients and controls (205 +/- 12% vs 239 +/- 15% of basal, respectively), but neutrophils from the most active patients (score > or = 3) increased CD11b/CD18 expression less than controls (175 +/- 12% of basal, P < 0.003, n = 9). The magnitude of the stimulated increment in expression of CD11b/CD18 on neutrophils correlated inversely with SLE activity (r = -0.64, P < 0.003, n = 20). Surprisingly, we observed no change in LS expression on neutrophils from SLE patients compared to controls (143 +/- 14 vs 141 +/- 16 RFU, respectively) even in patients with the highest activity indices (154 +/- 21 RFU). In contrast to CD11b/CD18, there was no correlation between LS expression and disease activity (r = 0.12, P = NS). Stimulation of neutrophils reduced the expression of LS similarly in both controls and SLE patients (67 +/- 3% vs 58 +/- 4% reduction, respectively) and did not correlate with disease activity (r = 0.07, P = NS, n = 20). These results show, for the first time, that changes in CD11b/CD18 expression do not correlate with LS expression on neutrophils from patients with active SLE.(ABSTRACT TRUNCATED AT 400 WORDS)
— id: 56565, year: 1994, vol: 71, page: 281, stat: Journal Article,

Immunocomplexes stimulate different signalling events to chemoattractants in the neutrophil and regulate L-selectin and beta 2-integrin expression differently
Molad Y; Haines KA; Anderson DC; Buyon JP; Cronstein BN
1994 May 1;299(Pt 3):881-887, Biochemical journal
Neutrophils express receptors for numerous phlogistons which, when occupied, trigger distinct signal-transduction pathways. Previous studies have shown that stimulation of neutrophils with chemoattractants induces shedding of the adhesive molecule L-selectin and increased expression of the beta 2-integrin CD11b/CD18. We determined the effect of ligation of classic, G-protein-linked chemoattractant receptors [C5a, interleukin-8 (IL-8), formylmethionyl-leucylphenylalanine (FMLP) and substance P], receptors for the Fc portion of IgG (Fc gamma receptors) and receptors for transforming growth factor beta (TGF beta) on expression of adhesive molecules by neutrophils and the stimulus-transduction mechanisms thought to mediate these changes. We were surprised to observe that occupancy of Fc gamma receptors by immunocomplexes (BSA-anti-BSA) stimulated increased expression by neutrophils of CD11b/CD18 at concentrations which did not affect L-selectin expression (EC50 9 micrograms/ml versus 350 micrograms/ml respectively, P < 0.00001, n = 5). In contrast, similar to previous studies, recombinant C5a, recombinant IL-8 and FMLP all stimulated increased expression of CD11b/CD18 (170-260% of basal, P < 0.001, n = 5) and shedding of L-selectin (56-75% reduction from basal, P < 0.001, n = 5) at similar concentrations and with similar potencies (EC50 = 2, 5, and 3 nM respectively). In contrast, neither TGF beta 1 nor, surprisingly, substance P affected expression of CD11b/CD18 or L-selectin. The regulation of expression of CD11b/CD18 or L-selectin in response to FMLP or immunocomplexes was unaffected by cytochalasin B (5 micrograms/ml) or the tyrosine kinase inhibitor tyrphostin-25 (25 microM). Although occupancy of both chemoattractant (FMLP) and Fc gamma receptors stimulated increments in the second messenger diacylglycerol, disruption of actin microfilaments by cytochalasin B enhanced diacylglycerol generation in response to FMLP but not in response to ligation of Fc gamma receptors. Moreover, both FMLP and immune aggregates provoked fluxes of intracellular Ca2+ concentration which differed with respect to both magnitude and kinetics and did not correlate well with regulation of adhesive-molecule expression. As upregulation of CD11b/CD18 is tightly linked to exocytosis of specific granules, these results suggest that shedding of L-selectin by activated neutrophils is not linked to exocytosis. These studies provide further evidence that receptors for chemoattractants and immunocomplexes on the neutrophil are linked to multiple signalling pathways
— id: 6462, year: 1994, vol: 299, page: 881, stat: Journal Article,

POSTNATAL TREATMENT OF COMPLETE CONGENITAL HEART-BLOCK WITH GLUCOCORTICOIDS
RIDER, LG; BUYON, JP; SHERRY, DD
1994 JUN ;37(6):R17-R17, Arthritis & rheumatism
— id: 52434, year: 1994, vol: 37, page: R17, stat: Journal Article,

The role of nitric oxide in the pathogenesis of preeclampsia
Seligman SP; Buyon JP; Clancy RM; Young BK; Abramson SB
1994 Oct;171(4):944-948, American journal of obstetrics & gynecology
OBJECTIVE: Nitric oxide, a potent vasodilator released by endothelial cells, inhibits platelet aggregation and adhesion to vascular endothelial surfaces. Because endothelial cell damage is considered pivotal in the pathogenesis of preeclampsia, this study was initiated to determine whether nitric oxide production is decreased in patients with preeclampsia. STUDY DESIGN: Twenty-six patients with preeclampsia (as defined by a blood pressure > or = 140 mm Hg systolic or 90 mm Hg diastolic plus proteinuria, > or = 300 mg per 24 hours or > or = 2+ by dipstick, both occurring on two occasions > or = 4 hours apart) and 26 normotensive women with singleton gestations in the third trimester were studied. Because nitric oxide is spontaneously oxidized to both nitrite and nitrate, two analytic assays were used serially. Serum nitrite levels were initially determined with the Greiss reagent and subsequently analyzed with Escherichia coli nitrate reductase. RESULTS: With the Greiss reagent alone the mean +/- SEM of serum nitrite level in 26 patients with preeclampsia was significantly decreased compared with 26 normotensive patients (3.46 +/- 1.43 mumol/L vs 4.65 +/- 0.85 mumol/L, p = 0.02). With the addition of the nitrate reductase enzyme of Escherichia coli the mean +/- SEM of serum nitrite level in 26 preeclamptic patients was again significantly decreased compared with 26 normotensive patients (20.04 +/- 1.25 mumol/L vs 27.38 +/- 2.23 mumol/L, p = 0.02). One patient with the syndrome of hemolysis, elevated liver enzymes, and low platelets demonstrated a concurrent decrease in serum nitrite over a 2-week period, emphasizing the relationship of nitric oxide to the pathophysiologic features of the syndrome. CONCLUSIONS: Circulating levels of nitrite are decreased in patients with preeclampsia. These data support the concept that diminished nitric oxide synthesis contributes to the pathophysiologic changes seen in preeclampsia
— id: 6747, year: 1994, vol: 171, page: 944, stat: Journal Article,

Autoantibody-associated congenital heart block: outcome in mothers and children
Waltuck J; Buyon JP
1994 Apr 1;120(7):544-551, Annals of internal medicine
OBJECTIVE: To determine the initial clinical status and the long-term outcome of mothers and their children with autoantibody-associated congenital heart block. DESIGN: Dynamic, longitudinal cohort study. PATIENTS: 55 children with isolated congenital heart block, their 52 mothers, and 5 other women currently carrying fetuses with congenital heart block. All maternal sera contained antibodies to SSA/Ro alone or to both SSA/Ro and SSB/La. MEASUREMENTS: Clinical information obtained from mailed questionnaires, telephone interviews, primary physicians, and chart reviews. RESULTS: When congenital heart block was identified in the children, 23 women were asymptomatic, 15 had systemic lupus erythematosus, 8 had the Sjogren syndrome, and 11 had an undifferentiated autoimmune syndrome. Follow-up ranged from 1 week to 20 years (median, 3.7 years). Eleven (48%) of the 23 initially asymptomatic mothers developed symptoms of a rheumatic disease (0.15 status changes/patient-year of follow-up; 6 (26%) developed an undifferentiated autoimmune syndrome, 2 (9%) developed the Sjogren syndrome, and 3 (13%) developed systemic lupus erythematosus. One mother with the Sjogren syndrome progressed to systemic lupus erythematosus. Four (16%) of 25 subsequent pregnancies in 22 women were complicated by heart block. Seventeen affected children died, 12 within 1 month of birth. Pacemakers were implanted in 37 (67%) of the 55 children, 27 within 3 months after birth. CONCLUSION: The development of rheumatic disease in asymptomatic mothers identified by the birth of a child with congenital heart block is common but not universal. The risk for congenital heart block in subsequent pregnancies is low. One third of the children with autoantibody-associated congenital heart block die in the early neonatal period and, of those who survive, most require pacemakers
— id: 12979, year: 1994, vol: 120, page: 544, stat: Journal Article,

UP-REGULATION OF ENDOTHELIAL-CELL ADHESION MOLECULES IN SYSTEMIC LUPUS-ERYTHEMATOSUS - THE SHWARTZMAN PHENOMENON REVISITED
BELMONT, HM; BUYON, J; GIORNO, R; ABRAMSON, SB
1993 APR ;41(2):A317-A317, Clinical research
— id: 54279, year: 1993, vol: 41, page: A317, stat: Journal Article,

NYU HJD LUPUS STUDY-GROUP CYCLOPHOSPHAMIDE (CY) EXPERIENCE - RETROSPECTIVE STUDY OF 51 PATIENTS
BELMONT, HM; STORCH, M; BUYON, J; ABRAMSON, S
1993 SEP ;36(9):S229-S229, Arthritis & rheumatism
— id: 52192, year: 1993, vol: 36, page: S229, stat: Journal Article,

Congenital complete heart block
Buyon JP
1993 Oct;2(5):291-295, Lupus
— id: 13067, year: 1993, vol: 2, page: 291, stat: Journal Article,

Identification of mothers at risk for congenital heart block and other neonatal lupus syndromes in their children. Comparison of enzyme-linked immunosorbent assay and immunoblot for measurement of anti-SS-A/Ro and anti-SS-B/La antibodies
Buyon JP; Winchester RJ; Slade SG; Arnett F; Copel J; Friedman D; Lockshin MD
1993 Sep;36(9):1263-1273, Arthritis & rheumatism
OBJECTIVE. To identify the fine specificity patterns of maternal anti-SS-A/Ro and anti-SS-B/La antibodies that are associated with the birth of a child with transient or permanent manifestations of neonatal lupus syndromes, and to suggest a predictor algorithm for use in counseling. METHODS. Sera were obtained from 4 groups of mothers: 57 whose children had congenital heart block, 12 whose children had transient dermatologic or hepatic manifestations of neonatal lupus but no detectable cardiac involvement, 152 with systemic lupus erythematosus and related autoimmune diseases, who gave birth to healthy infants, and 30 with autoimmune diseases whose pregnancy resulted in miscarriage, fetal death, or early postpartum death unrelated to neonatal lupus. Antibodies to SS-A/Ro and SS-B/La were assessed by enzyme-linked immunosorbent assay (ELISA) and by sodium dodecyl sulfate (SDS)-immunoblot. RESULTS. Anti-SS-A/Ro antibodies were identified by ELISA in 100%, 91%, 47%, and 43% of the mothers of infants with heart block, with transient neonatal lupus, healthy infants, and fetal death, respectively. High titers of anti-SS-A/Ro antibodies were present more often in mothers of children with cardiac disease or transient neonatal lupus than in either of the other 2 groups. Maternal antibodies to SS-B/La were detected by ELISA in 76% of the heart block group, 73% of the cutaneous neonatal lupus group, 15% of the group with healthy children, and 7% of the fetal death group. On SDS-immunoblot, sera from 91% of the heart block group mothers who had antibodies to SS-A/Ro but not to SS-B/La recognized at least 1 SS-A/Ro antigen, with significantly greater reactivity against the 52-kd component. In contrast, only 62% of the anti-SS-A/Ro positive, anti-SS-B/La negative responders in the healthy group recognized the 52-kd and/or the 60-kd component. Although there was no profile of anti-SS-A/Ro response unique to the mothers of children with heart block or cutaneous manifestations of neonatal lupus, only 1% of the healthy infants were born to mothers with antibodies directed to both the 52-kd SS-A/Ro and 48-kd SS-B/La antigens and not to the 60-kd SS-A/Ro antigen. CONCLUSION. Women with antibodies to both SS-A/Ro and SS-B/La have an increased risk of giving birth to children with neonatal lupus, especially if the anti-SS-A/Ro response identifies the 52-kd component on SDS-immunoblot. Women whose sera contain only anti-SS-A/Ro antibodies in low titer and only recognize determinants that are altered by conditions of SDS-immunoblot have a low risk for giving birth to a child with neonatal lupus. Specific antibody profiles do not distinguish among the manifestations of the neonatal lupus syndromes
— id: 57439, year: 1993, vol: 36, page: 1263, stat: Journal Article,

First international conference on rheumatic diseases in pregnancy
Buyon, J P; Yaron, M; Lockshin, M D
1993 Jan;36(1):59-64, Arthritis & rheumatism
— id: 73566, year: 1993, vol: 36, page: 59, stat: Journal Article,

PAST AND PRESENT USAGE OF ORAL-CONTRACE
BUYON, JP; ABRAMSON, S; BELMONT, HM
1993 SEP ;36(9):S229-S229, Arthritis & rheumatism
— id: 52193, year: 1993, vol: 36, page: S229, stat: Journal Article,

INTEGRIN CD11B/CD18 IS DIFFERENTIALLY PHOSPHORYLATED IN PLASMA VERSUS SPECIFIC GRANULE MEMBRANES OF NEUTROPHILS
BUYON, JP; PHILIPS, MR; SLADE, SG; LESZCZYNSKAPIZIAK, J; ABRAMSON, SB
1993 APR ;41(2):A377-A377, Clinical research
— id: 54285, year: 1993, vol: 41, page: A377, stat: Journal Article,

IDENTIFICATION OF 2 ANTIGENIC DETERMINANTS ON THE 52KD RO PROTEIN
BUYON, JP; SLADE, SG; REVEILLE, JD; HAMEL, J
1993 APR ;41(2):A317-A317, Clinical research
— id: 54278, year: 1993, vol: 41, page: A317, stat: Journal Article,

IMMUNOPRECIPITATION INCREASES IDENTIFICATION OF ANTI-LA ANTIBODIES IN ANTI-RO SERA
CALDWELL, KE; WALTUCK, J; CHAN, EKL; BUYON, JP
1993 SEP ;36(9):S239-S239, Arthritis & rheumatism
— id: 52194, year: 1993, vol: 36, page: S239, stat: Journal Article,

ONTOGENY OF COMPLEMENT REGULATORY PROTEINS - VARIED EXPRESSION IN DIFFERENT FETAL TISSUES
GORELICK, A; OGLESBY, T; RASHBAUM, W; SLADE, S; ATKINSON, J; BUVON, JP
1993 SEP ;36(9):S257-S257, Arthritis & rheumatism
— id: 52197, year: 1993, vol: 36, page: S257, stat: Journal Article,

IMMUNE-COMPLEXES (IC) AND CHEMOATTRACTANTS (IL-8, C5A AND FMLP) DIFFERENTIALLY MODULATE INTEGRIN AND SELECTIN EXPRESSION ON NEUTROPHILS (PMNS)
HAINES, KA; MOLAD, Y; ANDERSON, DC; BUYON, JP; CRONSTEIN, BN
1993 APR ;41(2):A278-A278, Clinical research
— id: 54273, year: 1993, vol: 41, page: A278, stat: Journal Article,

Treatment of neonatal lupus: case report and review of the literature
Rider, L G; Buyon, J P; Rutledge, J; Sherry, D D
1993 Jul;20(7):1208-1211, Journal of rheumatology
A critically ill infant with congenital complete heart block and neonatal lupus was treated with pulse steroids, exchange transfusion and intravenous gammaglobulin. Transient improvement in clinical status and laboratory results occurred, although the infant died. Based on this report and review of prior experience with immunosuppressive therapy, prenatal treatment of the mother with betamethasone or dexamethasone has been successful in resolving the cardiomyopathy/myocarditis associated with neonatal lupus. Postnatal treatment of the infant with steroids appears to be beneficial for persistent hepatic and hematologic manifestations. The evidence for exchange transfusion is less certain
— id: 73565, year: 1993, vol: 20, page: 1208, stat: Journal Article,

Neonatal hemochromatosis associated with maternal autoantibodies against Ro/SS-A and La/SS-B ribonucleoproteins
Schoenlebe, J; Buyon, J P; Zitelli, B J; Friedman, D; Greco, M A; Knisely, A S
1993 Oct;147(10):1072-1075, American journal of diseases of children
Liver disease in the fetus and neonate may be associated with maternal Sjogren's syndrome (neonatal lupus erythematosus). The infant of a mother with Sjogren's syndrome and high anti-Ro/SS-A and anti-La/SS-B antibody titers presented at 2 1/2 weeks of age with overwhelming hepatic insufficiency and died at 6 weeks of age. Autopsy confirmed the antemortem diagnosis of neonatal hemochromatosis. We discuss the possibility of a relationship between these two conditions
— id: 73564, year: 1993, vol: 147, page: 1072, stat: Journal Article,

APPROACH TO FETAL IDENTIFICATION AND THERAPY OF CONGENITAL HEART-BLOCK (CHB)
WALTUCK, J; COPEL, J; KLEINMAN, C; BUYON, JP
1993 SEP ;36(9):S97-S97, Arthritis & rheumatism
— id: 52190, year: 1993, vol: 36, page: S97, stat: Journal Article,

Activation of the complement pathway: comparison of normal pregnancy, preeclampsia, and systemic lupus erythematosus during pregnancy
Abramson SB; Buyon JP
1992 Oct-Dec;28(3-4):183-187, American Journal of Reproductive Immunology (1989)
To evaluate a flare of systemic lupus erythematosus (SLE) during pregnancy and to differentiate it from diseases of pregnancy, serological parameters are often utilized. However, there are conflicting reports regarding the merit of conventional measurements of complement and activation products. While in normal pregnancy the levels of serum C3, C4, and CH50 gradually rise, a decline in these levels occurs during the course of pregnancy in selected SLE patients. There is controversy regarding whether such falls represent decreases in the overall synthesis of complement or activation, the former theory being supported by a report of normal levels of the C1s-C1 inhibitor complex. During normal pregnancies, increases of complement split products, such as plasma C3a, may occur, and these correlate positively with elevations of C3. In pregnancies complicated by lupus, increases of C3a are often accompanied by a decline in total C3 and CH50. In a minority of non-SLE patients, preeclampsia has been associated with elevations of a variety of complement split products. Ba, C3a, C4d, SC5b-9, indicating activation of both the classical and alternative pathways. The CH50 levels tend to remain normal in these patients. In contrast, elevations of complement split products frequently accompany disease flares in patients with SLE. A high ratio of CH50/Ba may differentiate patients with preeclampsia from those with active SLE. A decline in conventional measures of C3, C4, or CH50 which is accompanied by elevations of complement split products appears to differentiate a lupus flare from non-SLE diseases of pregnancy
— id: 9744, year: 1992, vol: 28, page: 183, stat: Journal Article,

Anti-Ro/SS-A antibodies in the pathophysiology of congenital heart block in neonatal lupus syndrome, an experimental model. In vitro electrophysiologic and immunocytochemical studies
Alexander, E; Buyon, J P; Provost, T T; Guarnieri, T
1992 Feb;35(2):176-189, Arthritis & rheumatism
OBJECTIVE. To determine whether anti-Ro/SS-A antibodies selectively bind to neonatal cardiac cells and alter membrane repolarization. METHODS. An in vitro electrophysiologic and immunocytochemical experimental model contrasting neonatal and rabbit cardiac tissue was employed. RESULTS. Sera and IgG-enriched fractions from anti-Ro/SS-A antibody-positive mothers of infants with neonatal lupus erythematosus and congenital heart block bind to neonatal, rather than adult, rabbit cardiac tissue and alter the transmembrane action potential (i.e., inhibit repolarization). The additional presence of anti-La/SS-B antibodies was not additive or synergistic for these immunocytochemical and electrophysiologic features. Sera containing other antibody specificities (i.e., anti-native DNA, cardiolipin, Sm, and nuclear RNP) failed to stain the neonatal cardiac tissue or produced alterations in membrane repolarization. CONCLUSION. Anti-Ro/SS-A antibodies may play a pathophysiologic role in the development of congenital heart block in neonatal lupus
— id: 73568, year: 1992, vol: 35, page: 176, stat: Journal Article,

THE SHWARTZMAN PHENOMENON IN SYSTEMIC LUPUS-ERYTHEMATOSUS - A NOVEL MECHANISM OF VASCULAR INJURY
BELMONT, HM; BUYON, J; GIORNO, R; ABRAMSON, S
1992 SEP ;35(9):S211-S211, Arthritis & rheumatism
— id: 51851, year: 1992, vol: 35, page: S211, stat: Journal Article,

Neonatal lupus syndromes
Buyon JP
1992 Oct-Dec;28(3-4):259-263, American Journal of Reproductive Immunology (1989)
Neonatal lupus is a model of passively acquired autoimmunity in that immune abnormalities in the mother lead to the production of antibodies that cross the placenta and injure the developing fetus. Congenital complete heart block (CCHB), a permanent manifestation of neonatal lupus, is detectable after 18 wk gestation. Transient manifestations include cutaneous, hepatic, and hematologic abnormalities that occur at variable frequency. To date, there is a universal association of CCHB with maternal antibodies to SSA/Ro-SSB/La ribonucleoproteins, detectable by high ratio monomer:crosslinker SDS-immunoblot. Intriguingly, cardiac disease and often other manifestations are not present in the mother, raising the hypothesis that there is differential expression and/or accessibility of SSA/Ro-SSB/La antigens in fetal vs. adult tissues. CCHB may be a final consequence of a more widespread inflammatory response in the heart, including the existence of an associated myocarditis. In contrast to the in utero onset of CCHB, skin lesions generally become apparent after birth. Ultraviolet exposure may be an initiating factor and exacerbate an existing rash. Several studies have documented the predominance of DR3 alleles in mothers of affected offspring, frequently associated with the extended haplotype A1,B8. Available evidence suggests that fetal genetic differences in the major histocompatibility complex (MHC) do not influence susceptibility. The recommended clinical approach includes obstetric and rheumatologic management of both the fetus identified with CCHB and the fetus with a normal heart beat but at high risk of developing CCHB. Fetal echocardiogram is essential in diagnosing and following disease and may suggest the presence of an associated myocarditis.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 13434, year: 1992, vol: 28, page: 259, stat: Journal Article,

Assessment of disease activity and impending flare in patients with systemic lupus erythematosus. Comparison of the use of complement split products and conventional measurements of complement
Buyon JP; Tamerius J; Belmont HM; Abramson SB
1992 Sep;35(9):1028-1037, Arthritis & rheumatism
OBJECTIVE. To determine whether increased levels of the complement split products generated in the activation of the alternative or classical pathway accompany more severe disease activity in patients with systemic lupus erythematosus (SLE) and whether these measurements are useful in predicting flares of disease. METHODS. Levels of Ba, Bb, SC5b-9, and C4d were measured in 380 plasma samples obtained from 86 SLE patients who were prospectively followed up for 15 months. RESULTS. In the 20 patients who had inactive disease at the initiation of the study, the mean values of all of the complement split products at entry were within the normal range. In the 47 patients with stable or moderate disease activity, levels of Ba were significantly increased, while the mean values for Bb, SC5b-9, and C4d did not differ significantly from those in patients with inactive disease. The mean entry value of each analyte was highest in the group of 19 patients who had the most severe disease activity at initial evaluation. Traditional measurements of complement, i.e., C3, C4, and CH50, followed similar trends, but did not discriminate between the 3 groups of patients as well as did measurements of the split products. Analysis of the disease course in the patients with inactive or stable/moderate disease revealed that an elevated level of C4d had the most sensitivity with regard to subsequent flare, while an elevated Bb level had the highest specificity and the greatest predictive value. CONCLUSION. These data suggest that elevated levels of complement split products, particularly products of alternative and terminal pathway activation, more accurately reflect disease activity than do conventional measurements of complement in SLE and may be useful in the prediction of impending disease flares
— id: 9745, year: 1992, vol: 35, page: 1028, stat: Journal Article,

Activation of the alternative complement pathway accompanies disease flares in systemic lupus erythematosus during pregnancy
Buyon JP; Tamerius J; Ordorica S; Young B; Abramson SB
1992 Jan;35(1):55-61, Arthritis & rheumatism
OBJECTIVE. To assess the activity of systemic lupus erythematosus (SLE) during pregnancy and to distinguish it from preeclampsia. METHODS. We prospectively measured the complement activation products Ba, Bb, SC5b-9, and C4d, as well as the conventional complement determinants C3, C4, and CH50, during pregnancy in 14 patients with SLE and 10 women with preeclampsia. RESULTS. Four of the 14 SLE patients were considered to have disease flares, 3 occurring in the second trimester and 1 postpartum. In these patients, significant abnormalities of Ba, Bb, SC5b-9, and CH50 were noted. In contrast, measures of C4d did not distinguish between pregnant patients who had flares and those whose SLE remained stable. Although decreased values of C3 were rarely seen in the patients with stable disease, normal values of C3 during lupus pregnancy were not reliably associated with stable disease. Three of 10 non-SLE patients with preeclampsia had elevated levels of Ba; however, in each case, the CH50 level was close to or within the normal range. This was in sharp contrast to the findings observed in the 4 patients with active SLE, in whom high levels of plasma Ba were always associated with low CH50 values. Moreover, the ratio of CH50 to Ba was significantly lower in the patients with lupus flares than in the non-SLE patients with preeclampsia. CONCLUSION. While a decline in the CH50 level alone could otherwise be attributed to decreased synthesis of complement components, these data demonstrate that ongoing activation of the alternative complement pathway can accompany disease flares in pregnant women with SLE
— id: 9752, year: 1992, vol: 35, page: 55, stat: Journal Article,

DISSECTING THE ANTI-52KD RO RESPONSE IN COMPLETE HEART-BLOCK (CHB)
BUYON, JP; SLADE, S; REVEILLE, J; CHAN, EKL
1992 SEP ;35(9):S69-S69, Arthritis & rheumatism
— id: 51843, year: 1992, vol: 35, page: S69, stat: Journal Article,

MESSENGER-RNA AND PROTEIN EXPRESSION OF RO/LA IN HUMAN FETAL AND ADULT ORGANS
BUYON, JP; SLADE, SG; CHAN, EKL; WINCHESTER, RJ
1992 APR ;40(2):A222-A222, Clinical research
— id: 51982, year: 1992, vol: 40, page: A222, stat: Journal Article,

DIFFERENTIAL PHOSPHORYLATION OF CD11B/CD18 IN NEUTROPHIL SPECIFIC GRANULE AND PLASMA-MEMBRANES
BUYON, JP; SLADE, SG; LESZCYNSKAPIZIAK, J; PHILIPS, M; ABRAMSON, SB
1992 SEP ;35(9):S96-S96, Arthritis & rheumatism
— id: 51845, year: 1992, vol: 35, page: S96, stat: Journal Article,

COMPLETE HEART-BLOCK (CHB) - TITERS OF ANTI-RO/LA ANTIBODIES IN MATERNAL INFANT PAIRS
CRAWFORD, B; SLADE, S; COPEL, J; CHAN, EKL; BUYON, JP
1992 SEP ;35(9):S136-S136, Arthritis & rheumatism
— id: 51848, year: 1992, vol: 35, page: S136, stat: Journal Article,

PENTOXIFYLLINE AND PHOSPHATASE INHIBITORS MODULATE NEUTROPHIL AGGREGATION VIA DIFFERENT PATHWAYS
MERRILL, J; WINCHESTER, R; BUYON, JP
1992 APR ;40(2):A235-A235, Clinical research
— id: 51985, year: 1992, vol: 40, page: A235, stat: Journal Article,

INFLAMMATION AND NEUTROPHIL-ENDOTHELIAL INTERACTIONS - CHEMOATTRACTANTS AND IMMUNE-COMPLEXES DIFFERENTIALLY REGULATE EXPRESSION OF ADHESIVE MOLECULES ON NEUTROPHILS (PMNS)
MOLAD, Y; ANDERSON, DC; BUYON, JP; CRONSTEIN, BN
1992 SEP ;35(9):S96-S96, Arthritis & rheumatism
— id: 51844, year: 1992, vol: 35, page: S96, stat: Journal Article,

INTRAVASCULAR NEUTROPHIL (PMN) ACTIVATION IN SLE - DISSOCIATION BETWEEN INCREASED CD11B/CD18 (CR3) AND DECREASED L-SELECTIN (LS) EXPRESSION ON NEUTROPHILS (PMNS)
MOLAD, Y; BUYON, J; ANDERSON, DC; ABRAMSON, SB; CRONSTEIN, BN
1992 SEP ;35(9):S192-S192, Arthritis & rheumatism
— id: 51849, year: 1992, vol: 35, page: S192, stat: Journal Article,

Quantitative electroencephalography. A new approach to the diagnosis of cerebral dysfunction in systemic lupus erythematosus
Ritchlin CT; Chabot RJ; Alper K; Buyon J; Belmont HM; Roubey R; Abramson SB
1992 Nov;35(11):1330-1342, Arthritis & rheumatism
OBJECTIVE. Neuropsychiatric manifestations are common in patients with systemic lupus erythematosus (SLE), but accurate diagnosis is often difficult. We conducted a prospective study to determine the utility of neurometric quantitative electroencephalography (QEEG) as an indicator of cerebral dysfunction in SLE patients. METHODS. Fifty-two SLE patients were divided into 4 groups based on the results of neuropsychiatric evaluations. These included patients with objective evidence of neuropsychiatric SLE (NPSLE), patients with neuropsychiatric symptoms, patients with no evidence of NPSLE, and patients with a prior history of NPSLE. All QEEG findings were compared with data in an age-regressed normative database and with findings in an independent sample of normal subjects. RESULTS. QEEG sensitivity was 87%, and specificity was 75%. QEEG results were abnormal in 74% of the SLE patients with neuropsychiatric symptoms and in 28% of the patients with no evidence of active NPSLE. QEEG profiles varied as a function of the severity and type of neuropsychiatric manifestation present. Within this patient population, QEEG was more sensitive than magnetic resonance imaging, computed tomography scanning, or conventional EEG. CONCLUSION. Neurometric QEEG may be a sensitive indicator of cerebral dysfunction in patients with NPSLE and can differentiate patients with diverse neuropsychiatric manifestations. When combined with a careful clinical history and evaluation, QEEG provides information that may be useful for the early detection of NPSLE and for serial evaluation of disease activity and treatment efficacy
— id: 9743, year: 1992, vol: 35, page: 1330, stat: Journal Article,

Lupus anticoagulant activity of autoimmune antiphospholipid antibodies is dependent upon beta 2-glycoprotein I
Roubey, R A; Pratt, C W; Buyon, J P; Winfield, J B
1992 Sep;90(3):1100-1104, Journal of clinical investigation
It has been reported that antiphospholipid autoantibodies do not recognize phospholipid alone, but rather the plasma protein beta 2-glycoprotein I (beta 2GPI), or a beta 2GPI-phospholipid complex. In vitro beta 2GPI binds to anionic phospholipids and inhibits the prothrombinase activity of procoagulant membranes. In light of the fact that lupus anticoagulants, a type of antiphospholipid antibody, have similar anticoagulant properties, the relationship of beta 2GPI to lupus anticoagulant activity was investigated. IgG from patients with autoimmune diseases or syphilis were tested for anticardiolipin reactivity and lupus anticoagulant activity in the presence and absence of beta 2GPI. As expected, anti-cardiolipin reactivity associated with autoimmune disease was beta 2GPI dependent. In contrast, IgG from a patient with syphilis recognized cardiolipin alone and binding was inhibited by beta 2GPI. Autoimmune antiphospholipid antibodies prolonged the dilute Russell viper venom time of normal plasma, but had no effect on beta 2GPI-depleted plasma. Antiphospholipid antibodies associated with syphilis had no anticoagulant effect. RP-1, an anti-beta 2GPI mAb, had anticoagulant effects similar to those of autoimmune antiphospholipid antibodies. These data demonstrate that antiphospholipid autoantibodies exert lupus anticoagulant activity via an interaction with beta 2GPI. These antibodies and RP-1 appear to amplify the anticoagulant effect of beta 2GPI itself
— id: 73567, year: 1992, vol: 90, page: 1100, stat: Journal Article,

LUPUS ANTICOAGULANT ACTIVITY OF AUTOIMMUNE ANTIPHOSPHOLIPID ANTIBODIES IS DEPENDENT UPON BETA-2-GLYCOPROTEIN-I
ROUBEY, RAS; PRATT, CW; BUYON, JP; WINFIELD, JB
1992 APR ;40(2):A199-A199, Clinical research
— id: 51977, year: 1992, vol: 40, page: A199, stat: Journal Article,

NEONATAL AND MATERNAL OUTCOME IN COMPLETE HEART-BLOCK (CHB)
WALTUCK, J; BUYON, JP
1992 SEP ;35(9):S60-S60, Arthritis & rheumatism
— id: 51842, year: 1992, vol: 35, page: S60, stat: Journal Article,

Molecular definition and sequence motifs of the 52-kD component of human SS-A/Ro autoantigen
Chan EK; Hamel JC; Buyon JP; Tan EM
1991 Jan;87(1):68-76, Journal of clinical investigation
Serum SS-A/Ro autoantibodies are commonly found in patients with Sjogren's syndrome, systemic lupus erythematosus, neonatal lupus, and subacute cutaneous lupus. Two proteins of 60 and 52 kD have been described as targets for these autoantibodies. To define the 52-kD component unambiguously, cDNA clones were isolated from human HepG2 and MOLT-4 cell cDNA libraries. The identity of cDNA was established by (a) the specificity of the antibody affinity purified from the recombinant protein, (b) the reactivity of the purified recombinant protein with prototype SS-A/Ro sera in immunoblot and ELISA, and (c) two-dimensional gel comigration of MOLT-4 cell 52-kD protein and the recombinant protein. A 1.9-kb cDNA encoded the complete 52-kD protein containing 475 amino acids (Mr 54,082). Putative zinc-finger domains and a leucine zipper motif were identified in the amino-terminal half of the 52-kD protein, implicating its possible association with DNA/RNA. Sequence homology detected between the 52-kD protein and human ret transforming protein, and mouse T cell gene expression down-regulatory protein rpt-1, may provide leads to the functional role of the 52-kD protein in addition to the possibility that these proteins might constitute members of a subfamily of finger proteins
— id: 18583, year: 1991, vol: 87, page: 68, stat: Journal Article,

BETA-CHAIN PHOSPHORYLATION OF EACH NEUTROPHIL BETA-2 INTEGRIN IS INFLUENCED BY ITS ALPHA-CHAIN - OKADAIC ACID INDUCES INTEGRIN PHOSPHORYLATION BUT NOT LEUKOCYTE AGGREGATION
MERRILL, J; WINCHESTER, R; BUYON, JP
1991 APR ;39(2):A155-A155, Clinical research
— id: 51603, year: 1991, vol: 39, page: A155, stat: Journal Article,

Staurosporine inhibits neutrophil phagocytosis but not iC3b binding mediated by CR3 (CD11b/CD18)
Roubey, R A; Ross, G D; Merrill, J T; Walton, F; Reed, W; Winchester, R J; Buyon, J P
1991 May 15;146(10):3557-3562, Journal of immunology
C receptor CR3 (iC3b-receptor, CD11b/CD18) plays an essential role in several phagocytic and adhesive neutrophil functions. Recent evidence suggests that stimulus-induced phosphorylation of the CR3 beta-chain, CD18, may mediate certain neutrophil functions by transiently converting the molecule to an activated state. Staurosporine, a protein kinase C inhibitor that blocks PMA-induced CD18 phosphorylation, was used to study the functional relevance of this event. Neutrophils adhered to glass were assayed for binding and phagocytosis of iC3b-opsonized sheep E (EC3bi) in the presence or absence of PMA and/or staurosporine. Binding of EC3bi was markedly increased, not only by PMA, but also by staurosporine and by a combination of both agents (three- to sevenfold). The enhancement of rosetting by staurosporine was likely caused by increased surface expression of CR3 via exocytosis of specific granular contents. In contrast, staurosporine alone did not stimulate phagocytosis of EC3bi and markedly inhibited PMA-induced phagocytosis. Staurosporine also inhibited phagocytosis of yeast beta glucan particles, a CR3 ligand that, in contrast to EC3bi, is bound and ingested without additional prior treatment with PMA. beta glucan phagocytosis was associated with a low level of CD18 phosphorylation. Staurosporine did not block phagocytosis in general, because this agent had relatively little effect on FcR-mediated phagocytosis. These data demonstrate that phagocytosis mediated by CR3 requires activation of CR3 via a staurosporine-sensitive pathway. Increased binding of EC3bi, a function of increased surface expression of CR3, does not require activation of CR3 by such a pathway, confirming previous evidence for the independence of these two phenomena. A direct role for CD18 phosphorylation in the activation of CR3 for phagocytosis is consistent with these data
— id: 73569, year: 1991, vol: 146, page: 3557, stat: Journal Article,

Effective separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide by altering conditions of polyacrylamide gel electrophoresis
Buyon JP; Slade SG; Chan EK; Tan EM; Winchester R
1990 May 25;129(2):207-210, Journal of immunological methods
A method is described for the separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide. These two proteins anomalously co-migrate with the same relative mass under conditions of conventional sodium dodecyl sulfate polyacrylamide gel electrophoresis according to Laemmli using a stock solution of acrylamide: bisacrylamide 30:0.8, ratio = 37.5. A higher ratio of monomer to cross-linker, ratio = 172.4 used in a 15% acrylamide solution, readily separates the two peptide chains. This method facilitates the detection of the 52 kDa SSA/Ro component which otherwise might have been incorrectly assigned as a 48 kDa SSB/La polypeptide
— id: 18581, year: 1990, vol: 129, page: 207, stat: Journal Article,

Constitutive and induced phosphorylation of the alpha- and beta-chains of the CD11/CD18 leukocyte integrin family. Relationship to adhesion-dependent functions
Buyon JP; Slade SG; Reibman J; Abramson SB; Philips MR; Weissmann G; Winchester R
1990 Jan 1;144(1):191-197, Journal of immunology
We sought to determine whether the activation event which renders the CD11/CD18 leukocyte integrin/Leu-CAM glycoproteins capable of promoting cell to cell adhesion was associated with the induced posttranslational modification of phosphorylation. In neutrophils, two species of alpha-chains, a predominant CD11b 165-kDa subunit and a minor 150-kDa CD11c subunit were found to be constitutively phosphorylated. However, the 95-kDa CD18 common beta-chain was not phosphorylated in resting cells but became strongly phosphorylated in cells incubated with PMA. The beta-chain was phosphorylated in a dose-related manner within 1 min of the addition of PMA, reached maximal intensity between 4 to 10 min, and remained fully phosphorylated at 30 min. The similarities of the kinetics of homotypic aggregation induced by PMA to the time course of phosphorylation suggest that phosporylation may be relevant to at least this type of Leu-CAM-dependent adhesion. In contrast, in the presence of FMLP which induces aggregation with different kinetics than PMA, no phosphorylation of the common beta-chain was observed over a time interval of from 30 s to 10 min further emphasizing the apparent differences in the two modes of activation to an adhesive state. The phosphorylated species on neutrophils were readily detected by immunoprecipitation with each CD18 mAb and most but not all CD11b mAb which otherwise precipitated 125I-labeled CD11b species suggesting that the CD11b alpha-chain labelled with 32P may differ slightly from the 125I-labeled species in terms of its recognition by certain CD11b mAb. In mononuclear cells, similar constitutive phosphorylation of the CD11a and CD11c alpha-chains was observed that remained unchanged in the presence of either FMLP or PMA. As was demonstrated in neutrophils, phosphorylation of the CD18 beta-chains of mononuclear cells was not constitutive but was induced in the presence of PMA and not FMLP. Taken together these data suggest the existence of specific recognition sites on beta-chains for a regulatory kinase-phosphatase system
— id: 9762, year: 1990, vol: 144, page: 191, stat: Journal Article,

Systemic lupus erythematosus and the maternal-fetal dyad
Buyon, J P
1990 Apr;4(1):85-103, Bailliere's clinical rheumatology
Since systemic lupus erythematosus most frequently affects women of childbearing years, the management of patients during pregnancy is an important and common problem facing the clinician. This review concerns the effects of pregnancy on the course of maternal disease and fetal well-being. On the maternal side are the problems of renal disease which may exacerbate and be difficult to differentiate from pre-eclampsia especially when occurring in the third trimester. An active urinary sediment, falling C3 and CH50 and elevated complement split products of the alternative pathway and terminal attack complex may serve as useful parameters of lupus activity. In general, maternal disease is not an imposing threat and prospective studies suggest that the exacerbation rate is not significantly greater in the pregnant lupus patient than in the non-pregnant patient. On the fetal side are the problems of placental insufficiency and in utero attack on developing organs. Maternal antibodies such as those reactive with negatively charged phospholipids are associated with second trimester miscarriages and suggested, but not firmly established, thrombosis of placental vessels. The placental transfer of maternal antibodies against components of the rapidly expanding group of SSA/Ro-SSB/La ribonucleoproteins is strongly implicated in the transient and permanent manifestations of neonatal lupus. Using various techniques for defining the specificity of the antibody response most associated with heart block, the data suggest that mothers whose sera contain antibodies which recognize antigens of SSA/Ro-SSB/La on SDS-immunoblot are at greatest risk. In the absence of antibodies to SSB/La, mothers whose sera contain antibodies reactive only to bovine SSA/Ro by ELISA do not appear to be at high risk. A rational approach to in utero treatment of autoantibody mediated fetal myocarditis includes plasmapheresis and the use of dexamethasone. Finally, the safety of the commonly used medications for the treatment of lupus such as the nonsteroidal anti-inflammatory agents, glucocorticoids and anti-malarials during gestation and breast feeding, is addressed
— id: 73572, year: 1990, vol: 4, page: 85, stat: Journal Article,

Congenital complete heart block. A human model of passively acquired autoimmune injury
Buyon, J P; Winchester, R
1990 May;33(5):609-614, Arthritis & rheumatism
— id: 73570, year: 1990, vol: 33, page: 609, stat: Journal Article,

Molecular characterization and cloning of the 52 kDa SS-A/Ro protein
Chan EK; Hamel JC; Peebles CL; Buyon JP; Tan EM
1990 ;14(2-3):53-53, Molecular biology reports
— id: 18584, year: 1990, vol: 14, page: 53, stat: Journal Article,

Two pathways of CD11b/CD18-mediated neutrophil aggregation with different involvement of protein kinase C-dependent phosphorylation
Merrill JT; Slade SG; Weissmann G; Winchester R; Buyon JP
1990 Oct 15;145(8):2608-2615, Journal of immunology
The characteristics of homotypic neutrophil aggregation, mediated by the adhesion molecule CD11b/CD18, differ according to whether activation takes place via intracellular protein kinase C(PKC) inducers or chemoattractants. In response to diacylglycerol (DAG) analogues such as PMA and 1,2-dioctanoyl-sn-glycerol, a prolonged cellular aggregation occurs that is associated with intense phosphorylation of the CD18 beta-chain. In response to the chemoattractant FMLP, a more transient aggregation event results that is associated with minimal beta-chain phosphorylation. By using the PKC inhibitor staurosporine, we now show that these differences are likely to reflect two different pathways of activation. Both aggregation and phosphorylation induced by DAG analogues are completely abolished by staurosporine in a parallel dose-dependent manner. Conversely, FMLP-induced aggregation is enhanced and prolonged by staurosporine whereas the associated minimal phosphorylation event is further diminished by staurosporine. Accordingly, activation of neutrophil aggregation by DAG analogues is associated with and presumably due to phosphorylation of the CD18 beta-chain. This intense phosphorylation occurs via a staurosporine-sensitive kinase such as PKC. FMLP, on the other hand, appears to activate CD11b/CD18 by a distinct mechanism. This latter mechanism does not seem to be dependent on what may be a minor PKC-induced phosphorylation of the beta-chain, and indeed is enhanced by inhibition of PKC. Of note, staurosporine was also found to cause selective release of specific granules with concomitant increase in surface display of CD11b/CD18. These data further support previous observations that up-regulation of this adhesive molecule is not the primary event in the induction of cellular adhesiveness
— id: 14314, year: 1990, vol: 145, page: 2608, stat: Journal Article,

2 PATHWAYS OF CD11B/CD18 MEDIATED NEUTROPHIL AGGREGATION WITH DIFFERENT INVOLVEMENT OF PROTEIN KINASE-C (PKC) DEPENDENT PHOSPHORYLATION
Merrill, JT; Slade, SG; Winchester, R; Buyon, JP
1990 Apr;38(2):A364-A364, Clinical research
— id: 31961, year: 1990, vol: 38, page: A364, stat: Journal Article,

NEUTROPHIL CR3-MEDIATED PHAGOCYTOSIS BUT NOT IC3B BINDING REQUIRES PROTEIN-KINASE C-DEPENDENT PHOSPHORYLATION
Roubey, RAS; Ross, GD; Winchester, RJ; Buyon, JP
1990 Apr;38(2):A232-A232, Clinical research
— id: 31951, year: 1990, vol: 38, page: A232, stat: Journal Article,

Serum activity that confers acid lability to alpha-interferon in systemic lupus erythematosus: its association with disease activity and its independence from circulating alpha-interferon
Yee, A M; Yip, Y K; Fischer, H D; Buyon, J P
1990 Apr;33(4):563-568, Arthritis & rheumatism
We conducted a longitudinal evaluation of a patient with systemic lupus erythematosus who constitutively exhibited elevated levels of circulating alpha-interferon (alpha-IFN). This study demonstrated that the serum levels of an activity that renders the endogenous alpha-IFN acid labile are positively correlated with disease activity. This IFN acid lability-inducing activity can also be found in the sera of systemic lupus erythematosus patients who have active disease but who do not have circulating alpha-IFN
— id: 73571, year: 1990, vol: 33, page: 563, stat: Journal Article,

Anti-SS-A/Ro SS-B/La antibodies bind to neonatal rabbit cardiac cells and preferentially inhibit in vitro cardiac repolarization
Alexander, E L; Buyon, J P; Lane, J; Lafond-Walker, A; Provost, T T; Guarnieri, T
1989 Aug;2(4):463-469, Journal of autoimmunity
The neonatal lupus syndrome is the most common cause of isolated congenital heart block. There is, at present, little information about the putative role of anti-SS-A/Ro SS-B/La antibodies in the pathogenesis of congenital heart block in the neonatal lupus syndrome. Using an in vitro experimental model, the present study was designed to test the hypothesis that IgG antibodies in the sera of anti-SS-A/Ro SS-B/La-positive mothers of infants with isolated congenital heart block bind to and affect the transmembrane action potential of rabbit cardiac tissue. The results demonstrate a preferential inhibition of membrane repolarization (ADP-50 and ADP-90) and staining of cardiac cells within the neonatal, in contrast to the adult, rabbit heart by sera and IgG-enriched fractions from anti-SS-A/Ro SS-B/La-positive individuals. The results of the electrophysiologic studies demonstrate a pathophysiologic role for the IgG fraction of anti-SS-A/Ro SS-B/La-positive maternal sera in inhibiting neonatal rabbit cardiac repolarization. It is possible that antibodies to similar determinants expressed on the cell membrane of cardiac-conducting cells also may play a pathophysiologic role in the development of idiopathic congenital heart block in humans
— id: 73573, year: 1989, vol: 2, page: 463, stat: Journal Article,

Neonatal lupus and congenital complete heart block: manifestations of passively acquired autoimmunity
Buyon JP
1989 Sep-Oct;7 Suppl 3:S199-S203, Clinical & experimental rheumatology
The neonatal lupus syndromes, which comprise transient hematologic and cutaneous disorders as well as the permanent manifestation of heart block, are considered to result from injury by passively acquired maternal autoantibodies. The active placental transport of maternal IgG antibodies becomes operative late in the second trimester coincident with the time at which bradycardia and myocarditis become evident. Surprisingly there are no clinically detectable abnormalities in the maternal hearts. The recognition that antibodies to the SSA/Ro-SSB/La ribonucleoprotein complex were found in 85% of sera from mothers of offspring with neonatal lupus was an important advance and directed attention to these antigens as potential candidates despite their intracellular location. In the present review we describe an experimental approach to the treatment of a fetus diagnosed by in utero echocardiogram to have congenital complete heart block and to the prevention of this condition in an at-risk pregnancy. In an attempt to more specifically define the relevant antigen-antibody systems involved in the pathogenesis of neonatal lupus we have utilized the technique of immunoblot to evaluate sera from mothers of offspring with permanent manifestations of neonatal lupus including heart block and hepatic fibrosis.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 10504, year: 1989, vol: 7 Suppl 3, page: S199, stat: Journal Article,

Acquired congenital heart block. Pattern of maternal antibody response to biochemically defined antigens of the SSA/Ro-SSB/La system in neonatal lupus
Buyon JP; Ben-Chetrit E; Karp S; Roubey RA; Pompeo L; Reeves WH; Tan EM; Winchester R
1989 Aug;84(2):627-634, Journal of clinical investigation
The molecular basis of autoantibody reactivity with components of the SSA/Ro-SSB/La particle exhibited by sera of mothers of infants with severe and permanent manifestations of neonatal lupus (NLE) was investigated using immunoblotting and immunoprecipitation. The characteristics of NLE that were studied included congenital complete heart block (CCHB), second degree heart block, and hepatic fibrosis. Antibodies specific for one or more components of the SSA/Ro-SSB/La particle were found in sera from all 20 mothers of permanently affected infants. However, no antibody specific for a single peptide of this particle was common to all sera. Using tissue extracts from a human cell substrate, 80% of these sera had antibodies to one or more components of the SSA/Ro particle demonstrable by immunoblotting. The predominant antibody response in the NLE group was to the newly recognized 52-kD SSA/Ro peptide component. In contrast, antibodies to the 60-kD SSA/Ro component although present, were the least represented and not significantly increased in frequency among mothers of these infants, compared with a group of 31 mothers with autoimmune diseases such as systemic lupus erythromatosus (SLE) but who had healthy offspring. Antibodies directed to the 48-kD SSB/La antigen were demonstrated in 90% of the NLE mothers often accompanying antibodies against the 52-kD SSA/Ro component. The combination of antibodies to 48- and 52-kD structures was significantly increased in the NLE group, with an odds ratio of 35. The type of cell or tissue substrate was shown to influence detectability of antibodies. The 52-kD SSA/Ro peptide and the 48-kD SSB/La peptide were abundant in cardiac tissues from fetuses aged 18-24 wk, further supporting the possible relevance of these peptides to heart block
— id: 10526, year: 1989, vol: 84, page: 627, stat: Journal Article,

ASSOCIATION OF CONGENITAL COMPLETE HEART-BLOCK WITH MATERNAL ANTIBODY-RESPONSE TO THE NOVEL 52KD SSA/RO
Buyon, JP; Benchetrit, E; Karp, S; Roubey, RAS; Pompeo, L; Reeves, WH; Tan, EM; Winchester, RJ
1989 Apr;37(2):A506-A506, Clinical research
— id: 31706, year: 1989, vol: 37, page: A506, stat: Journal Article,

PHOSPHORYLATION OF THE ALPHA-CHAINS AND BETA-CHAINS OF THE CD11/CD18 INTEGRIN FAMILY - CONSTITUTIVE AND ACTIVATED STATES
BUYON, JP; REIBMAN, J; ABRAMSON, SB; PHILIPS, M; SLADE, SG; WEISSMANN, G; WINCHESTER, RJ
1989 APR ;37(2):A425-A425, Clinical research
— id: 51439, year: 1989, vol: 37, page: A425, stat: Journal Article,

Interferon alpha associated with systemic lupus erythematosus is not intrinsically acid labile
Yee, A M; Buyon, J P; Yip, Y K
1989 Mar 1;169(3):987-993, Journal of experimental medicine
The physicochemical properties of apparently acid-labile IFN-alpha from patients with SLE have been studied. The antigenicity, apparent molecular size, and isoelectric point of SLE IFN-alpha are indistinguishable from those of conventional, previously characterized, acid-stable subspecies of IFN-alpha. However, after partial purification by anion-exchange chromatography, SLE IFN-alpha no longer exhibits acid lability, suggesting that other plasma factor(s) are responsible for the acid lability of SLE IFN-alpha. Addition of SLE plasma, but not normal plasma, to conventional acid-stable IFN-alpha renders the exogenous IFN-alpha acid labile. Preliminary results demonstrate that an acid-dependent IFN-inactivating activity can be partially purified from SLE plasma by anion-exchange chromatography
— id: 73574, year: 1989, vol: 169, page: 987, stat: Journal Article,

Complete congenital heart block: risk of occurrence and therapeutic approach to prevention
Buyon J; Roubey R; Swersky S; Pompeo L; Parke A; Baxi L; Winchester R
1988 Jul;15(7):1104-1108, Journal of rheumatology
A retrospective literature review revealed that in 41% of cases of complete congenital heart block (CCHB) there was at least one other affected sibling, emphasizing the considerable risk of carrying a second affected fetus with CCHB. Therefore an aggressive approach was taken to prevent CCHB in a fetus with a high risk for the condition as defined by (1) presence of DR3 and high titers of antibodies to SSA(Ro) and SSB(La) in the mother (2) previous history of CCHB in a sibling. In a feasibility study, thrice weekly plasmapheresis was initiated in the 19th week of gestation to remove antibodies from the maternal circulation in advance of major placental transport to the developing fetus. Prednisone was also administered to decrease antibody synthesis. The concentration of total maternal IgG and antibodies to SSA(Ro) and SSB(La) were decreased by greater than 60% during the course of therapy. Planned delivery of a healthy baby was done at 36 weeks
— id: 11041, year: 1988, vol: 15, page: 1104, stat: Journal Article,

Dissociation between increased surface expression of gp165/95 and homotypic neutrophil aggregation
Buyon JP; Abramson SB; Philips MR; Slade SG; Ross GD; Weissmann G; Winchester RJ
1988 May 1;140(9):3156-3160, Journal of immunology
Whether homotypic neutrophil aggregation depends on the quantitative increase of gp165/95 molecules (Mac 1, CR3) recruited to the cell surface during activation was studied using mAb of the CD11b group that recognize distinct epitopes encoded by the alpha-subunit of this glycoprotein. After the addition of antibody MN41, neutrophils did not aggregate in response to a chemoattractant, FMLP. Blockade of preexisting surface gp165/95 by mAb MN41, followed by removal of the excess antibody from the mixture, was used to show that the molecules of gp165/95 newly expressed in response to stimulation by a chemoattractant were incapable of effectively mediating the induced cell-cell interactions of aggregation. Flow cytometry studies confirmed that binding of unlabeled antibody MN41 did not block further increases in surface expression of gp165/95 after stimulation with FMLP. These data suggest that molecules of gp165/95 exhibit two functionally distinct forms, one, present on the surface of freshly isolated neutrophils, that becomes competent to mediate the aggregation response upon activation by a stimulus and a second form that can be translocated to the cell surface by the stimulus but is greatly diminished if not lacking in the ability to participate in that aggregation event
— id: 9768, year: 1988, vol: 140, page: 3156, stat: Journal Article,

Surface expression of Gp 165/95, the complement receptor CR3, as a marker of disease activity in systemic Lupus erythematosus
Buyon JP; Shadick N; Berkman R; Hopkins P; Dalton J; Weissmann G; Winchester R; Abramson SB
1988 Jan;46(1):141-149, Clinical immunology & immunopathology
Complement-derived peptides capable of activating neutrophils appear in plasma during flares of systemic lupus erythematosus (SLE). One possible consequence of such activation is an increased expression of the surface adhesion promoting heterodimer gp165/95 (the complement receptor CR3). The quantity of gp165/95 was measured by indirect immunofluorescence using a monoclonal antibody of the CD11b group. Mol, directed to the alpha chain. Eighty-three percent of 26 patients with SLE expressed gp165/95 on their neutrophil surface to a greater extent than normals. The highest levels of surface gp165/95 were found in patients with the most severe disease, who also had the highest levels of the circulating anaphylatoxin C3a (mean = 560 ng/ml versus 147 ng/ml in controls). There was a negative correlation between expression of gp165/95 and absolute neutrophil count. Five individuals followed serially demonstrated an increase in surface gp165/95 during disease flares which returned to normal with clinical improvement. These data support the hypothesis that the neutrophils of patients with active SLE recruit increased numbers of gp165/95 molecules to their surface in respose to complement activation; these activated neutrophils bearing increased numbers of adhesion promoting gp165/95 may contribute to endothelial injury in SLE
— id: 9770, year: 1988, vol: 46, page: 141, stat: Journal Article,

DETERMINATION OF PLASMA COMPLEMENT SPLIT PRODUCTS (CSP) IN LUPUS IS THE MOST SENSITIVE PREDICTOR OF DISEASE-ACTIVITY
Buyon, J; Tamerius, J; Kolb, W; Morrow, P; Winchester, R; Weissmann, G; Abramson, S
1988 Apr;36(3):A531-A531, Clinical research
— id: 31505, year: 1988, vol: 36, page: A531, stat: Journal Article,

DIFFERENTIAL PHOSPHORYLATION OF THE ALPHA-CHAINS AND BETA- CHAINS OF THE LFA FAMILY GP165/95, P150/95 AND LFA-1
Buyon, JP; Abramson, SB; Slade, SG; Philips, M; Reibman, J; Weissmann, G; Winchester, RJ
1988 Sep;36(5):A799-A799, Clinical research
— id: 31447, year: 1988, vol: 36, page: A799, stat: Journal Article,

RECOGNITION OF A 48KDA COMPONENT BY SERA FROM MOTHERS OF OFFSPRING WITH CONGENITAL COMPLETE HEART-BLOCK IN WESTERN BLOT ANALYSIS
Buyon, JP; Karp, SW; Roubey, RAS; Winchester, RJ
1988 Apr;36(3):A531-A531, Clinical research
— id: 31504, year: 1988, vol: 36, page: A531, stat: Journal Article,

Increased levels of plasma anaphylatoxins in systemic lupus erythematosus predict flares of the disease and may elicit vascular injury in lupus cerebritis
Hopkins P; Belmont HM; Buyon J; Philips M; Weissmann G; Abramson SB
1988 May;31(5):632-641, Arthritis & rheumatism
We measured levels of complement anaphylatoxin split products, C3a and C5a, in the circulation of patients with systemic lupus erythematosus (SLE). In 23 SLE patients who were followed serially, the mean C3a value was 179 ng/ml during stable disease and 550 ng/ml during a disease flare. In 10 patients, C3a levels predicted disease activity, with the C3a value rising from a mean of 183 ng/ml at a time of stable disease to a mean of 242 ng/ml 1-2 months prior to a clinical exacerbation of disease. The mean C3a level in 5 patients with acute dysfunction of the central nervous system (CNS) was 1,297 ng/ml, which is significantly higher than that observed in patients with active disease but without CNS involvement (P less than 0.01). C5a levels were also significantly elevated in 4 patients with acute CNS disease. Pathologic specimens from 2 patients who died during an acute lupus flare revealed neutrophils occluding the cerebral and intestinal vessels. Fluorescein angiography in a patient with CNS lupus revealed vasoocclusive retinopathy. In 5 of 7 SLE patients who were pregnant, C3a levels were elevated, with a group mean value of 310 ng/ml. There was a negative correlation (r = -0.59) between C3a and C3 levels in pregnant patients with SLE, and this finding is consistent with complement activation as the cause of decreasing C3 levels. We suggest that serial measurements of C3a can predict flares of disease in lupus patients and can demonstrate complement activation during pregnancy in women with SLE. In addition, release of C3a and C5a (mediators of inflammation) into the circulation may elicit vascular injury, particularly in patients with lupus cerebritis
— id: 9769, year: 1988, vol: 31, page: 632, stat: Journal Article,

Up-regulation of the iC3b receptor (CR3) is neither necessary nor sufficient to promote neutrophil aggregation
Philips MR; Buyon JP; Winchester R; Weissmann G; Abramson SB
1988 Aug;82(2):495-501, Journal of clinical investigation
The iC3b receptor (CR3) is required for neutrophil adhesive functions, including homotypic aggregation. Because stimuli that enhance neutrophil adhesion also induce up-regulation of surface CR3, it is widely held that these two responses are causally related. We have dissociated CR3 display (immunofluorescence) from CR3 function (aggregation). Neutrophils isolated at 4 degrees C and rewarmed to 37 degrees C up-regulated surface CR3 twofold, but did not aggregate. The kinetics of FMLP-induced CR3 up-regulation were discordant with those of aggregation. In the absence of extracellular divalent cations, CR3 expression increased twofold after exposure to FMLP, but neutrophils did not aggregate. FMLP elicited 3.5-fold more aggregation than the ionophore A23187, yet less than one-half as much CR3 up-regulation. 3 mM sodium salicylate inhibited aggregation 55 +/- 4%, but had no effect on CR3 up-regulation. Conversely, 1 mM tetracaine completely inhibited CR3 up-regulation, while significantly enhancing aggregation. Neutroplasts expressed CR3, but did not up-regulate the receptor; in contrast, FMLP induced CR3-dependent aggregation of neutroplasts. We conclude that, although constitutive surface CR3 is required for neutrophil aggregation, the up-regulation of CR3 is neither necessary nor sufficient to promote cell-cell adhesion
— id: 9767, year: 1988, vol: 82, page: 495, stat: Journal Article,

Complement activation and vascular injury in systemic lupus erythematosus
Abramson S; Belmont HM; Hopkins P; Buyon J; Winchester R; Weissmann G
1987 Jun;14 Suppl 13:43-46, Journal of rheumatology. Supplement
The deposition of immune complexes within blood vessel walls results in the potential for complement activation and the release of chemotactic factors, such as fragments of C5 (C5fr). The generation of C5fr results in the intravascular aggregation of neutrophils with subsequent leukostatic occlusion of the pulmonary arterioles. The generation of C5fr may contribute to the pathogenesis of adult respiratory distress syndrome and other diseases. Studies were undertaken to determine the role of circulating complement derived peptides and intravascular neutrophil activation in systemic lupus erythematosus
— id: 59685, year: 1987, vol: 14 Suppl 13, page: 43, stat: Journal Article,

Intrauterine therapy for presumptive fetal myocarditis with acquired heart block due to systemic lupus erythematosus. Experience in a mother with a predominance of SS-B (La) antibodies
Buyon, J P; Swersky, S H; Fox, H E; Bierman, F Z; Winchester, R J
1987 Jan;30(1):44-49, Arthritis & rheumatism
An experimental therapeutic regimen for congenital lupus erythematosus, which consists of treating the mother with high doses of dexamethasone and performing plasmapheresis (3 times per week), was developed. This regimen was administered to a woman whose pregnancy was complicated by the abrupt onset of fetal pericarditis, myocarditis, and complete heart block in the twenty-fourth week of gestation. The effectiveness of plasmapheresis was evidenced by a decrease in the very high titer of SS-B (La) antibodies in the mother. After 5 weeks of therapy, the cardiac disease was ameliorated. The infant tolerated delivery at week 31, and the heart block has persisted. The rationale for this therapy for fetal disease is discussed, with emphasis on 2 goals: to diminish the quantity of maternal antibody that is transmitted to the fetus, and to suppress the manifestations of fetal carditis with the use of glucocorticoids that are not inactivated by the placenta. Tentative recommendations for more intensive prenatal monitoring of maternal and fetal status for those at risk are outlined
— id: 73575, year: 1987, vol: 30, page: 44, stat: Journal Article,

DISSOCIATION BETWEEN THE PHENOMENON OF GP165/95 (CR3) MEDIATED GRANULOCYTE AGGREGATION AND THE QUANTITY OF GP165/95 EXPRESSED ON THE CELL-SURFACE - EVIDENCE FOR 2 FUNCTIONALLY DISTINCT FORMS OF GP165/95
Buyon, JP; Phillips, M; Dalton, J; Abramson, S; Weissmann, G; Winchester, R
1987 Apr;35(3):A469-A469, Clinical research
— id: 31369, year: 1987, vol: 35, page: A469, stat: Journal Article,

AGGREGATION OF HUMAN-NEUTROPHILS IN RESPONSE TO CHEMOATTRACTANT BEARS NO RELATIONSHIP TO DISPLAY OF COMPLEMENT RECEPTOR (CR3)
Philips, M; Buyon, J; Hopkins, P; Gude, D; Winchester, R; Abramson, S; Weissmann, G
1987 Mar 1;46(3):405-405, Federation Proceedings (Federation of American Societies for Experimental Biology)
— id: 31260, year: 1987, vol: 46, page: 405, stat: Journal Article,

UP-REGULATION OF IC3B RECEPTORS IS NEITHER NECESSARY NOR SUFFICIENT FOR AGGREGATION OF NEUTROPHILS BY A CHEMOATTRACTANT
Philips, M; Buyon, J; Winchester, R; Weissmann, G; Abramson, S
1987 Apr;35(3):A486-A486, Clinical research
— id: 31193, year: 1987, vol: 35, page: A486, stat: Journal Article,

SURFACE EXPRESSION OF CR3 ON NEUTROPHILS (PMN) AS A MARKER OF DISEASE-ACTIVITY IN SYSTEMIC LUPUS-ERYTHEMATOSUS (SLE)
ABRAMSON, S; BUYON, J; SHADDICK, N; BERKMAN, R; HOPKINS, P; DALTON, J; WEISSMANN, G; WINCHESTER, R
1986 APR ;34(2):A614-A614, Clinical research
— id: 41405, year: 1986, vol: 34, page: A614, stat: Journal Article,

Serum complement values (C3 and C4) to differentiate between systemic lupus activity and pre-eclampsia
Buyon JP; Cronstein BN; Morris M; Tanner M; Weissmann G
1986 Aug;81(2):194-200, American journal of medicine
It is often difficult to differentiate between an exacerbation of systemic lupus erythematosus (SLE) and intercurrent pre-eclampsia in a patient with SL