Circulating anticentromere CENP-A and CENP-B antibodies in patients with diffuse and limited systemic sclerosis, systemic lupus erythematosus, and rheumatoid arthritis.

OBJECTIVE:To determine the disease sensitivity and specificity of testing for autoantibodies against 2 of the 3 main human centromere antigenic components, CENP-A and CENP-B (recombinant, expressed in baculovirus). METHODS:ELISA with CENP-A and CENP-B antigens were used to test 45 sera showing a centromere pattern by immunofluorescence (IFA) and sera from 96 patients with systemic sclerosis (SSc), subdivided into diffuse (dSSc) and limited (lSSc) forms. For controls, the same tests were performed on sera from 100 patients with rheumatoid arthritis (RA), 100 with systemic lupus erythematosus (SLE), and 50 random blood donors. Sera from all the patients with SSc were also tested for the presence of anti-Scl70 antibody by ELISA (bovine antigen), and for pattern and titer by IFA (HEp-2 cells). RESULTS:Of the 45 IFA positive sera, 93% were positive for anti-CENP-A and 91% for anti-CENP-B. There was a very good quantitative correlation between the antibody levels against these 2 centromere components (r = 0.597; p<0.001). Anti-CENP-A and B were found in 48% of patients with lSSc, and in 11% and 9%, respectively, of those with dSSc. The difference in the frequency of anti-CENP-A between the 2 patient groups was significant (chi-squared, p<0.001). Similar levels of anticentromere staining pattern by IFA were observed for these 2 groups. Anti-Scl70 was elevated in 8% of lSSc and 25% of dSSc patients; this difference was also significant (chi-squared, p = 0.02). Neither CENP-A nor CENP-B reacted with IgG from SSc patients containing anti-Scl70. The frequency of abnormal levels in patients with SLE and RA was, respectively, 11% and 3% for anti-CENP-A and 4% and 3% for anti-CENP-B. The reaction of IgG from SLE and RA patients with CENP-A was not inhibited by histone H3, i.e., it was not due to recognition of the histone-like domain in CENP-A. Thus, when 96 SSc patients were compared to 200 patients with RA and SLE, the disease specificity of anti-CENP-A and B was 93% and 96.5%, respectively. CONCLUSION:In addition to IFA, ELISA tests for CENP-A and CENP-B yield results with similar sensitivity and specificity for the diagnosis of SSc. CENP-A and CENP-B are primarily associated with lSSc. In SSc the autoantibody response is directed simultaneously and with similar amplitude against these 2 components of the centromere structure, whereas in other autoimmune diseases the response is directed mainly against one of the 2 components.

Russo K ，Hoch S ，Dima C ，Varga J ，Teodorescu M ... -  《JOURNAL OF RHEUMATOLOGY》

Anti-CENP-B response in sera of uranium miners exposed to quartz dust and patients with possible development of systemic sclerosis (scleroderma).

Conrad K ，Stahnke G ，Liedvogel B ，Mehlhorn J ，Barth J ，Blasum C ，Altmeyer P ，Sönnichsen N ，Frank KH ... -  《JOURNAL OF RHEUMATOLOGY》

[Clinical significance of anti-centromere antibody and anti-CENP-B antibody in sera of patients with primary biliary cirrhosis].

Onozuka Y ，Shibata M ，Yonezawa H ，Terauti K ，Miyachi K ，Ueno Y ... -  《-》

Anticentromere autoantibodies in patients without Raynaud's disease or systemic sclerosis.

Anticentromere autoantibodies (ACA) are associated with Raynaud's disease and systemic sclerosis (SSc). ACA usually bind at least one of three major centromere proteins (CENPs), particularly CENP-B. We identified 16 patients with ACA who do not have Raynaud's disease or SSc. The objective of this study was to determine whether these 16 ACA differ in antigenic specificity from the ACA found in patients with Raynaud's disease or SSc. Binding of these serum ACA was tested using competition experiments with recombinant CENP-B, and native centromere proteins from HEp-2 cells and HeLa nuclear extracts in ELISAs, immunoblots, and indirect immunofluorescence assays. The ACA from these 16 patients are strikingly different from those obtained from patients who have Raynaud's disease or SSc. Only 5 of the 16 index sera (31.25%) bound CENP-B from two or more different sources by at least two methods. Six of these 16 sera (37.5%) did not bind CENP-B on ELISA, and 8 of 16 (43.75%) did not bind CENP-B on immunoblots. Three sera did not bind CENP-B either by ELISA or immunoblots. Of the 13 sera that bound CENP-B, their patterns of binding to CENP-B strongly suggested that they bind different epitopes within the CENP-B antigen. Independently of their binding to CENP-B, these sera reacted mainly with minor CENP antigens detected by HeLa nuclear extracts. We have identified unusual ACA not associated with Raynaud's disease or SSc.

Vázquez-Abad D ，Grodzicky T ，Senécal JL 《CLINICAL IMMUNOLOGY》

Anti-centromere antibodies in a large cohort of systemic sclerosis patients: comparison between immunofluorescence, CENP-A and CENP-B ELISA.

Anti-centromere antibodies (ACA) are useful biomarkers in the diagnosis of systemic sclerosis (SSc) where they are found in 20-40% of patients and, albeit with lower prevalence, in patients with other systemic autoimmune rheumatic diseases. Historically, ACA were detected by indirect immunofluorescence (IIF) on HEp-2 cells and confirmed by immunoassays using recombinant CENP-B. During the last few years, to accommodate high throughput diagnostics, a number of laboratories changed from IIF to ELISA assays. The objective of this study was to compare the detection of ACA by IIF to CENP-A and a CENP-B ELISA in a large cohort of SSc patients. Sera collected from SSc patients (n=834) were tested for ACA by IIF on HEp-2 cells (ImmunoConcepts, Sacramento, CA) and CENP-A and CENP-B ELISA (both Dr. Fooke Laboratorien GmbH, Neuss, Germany). Furthermore, other autoantibodies were determined by QUANTA-Plex(TM) SLE 8 profile (INOVA, San Diego, CA), QUANTA Lite RNA Pol III (INOVA) and PM1-Alpha ELISA (Dr. Fooke). The prevalence of ACA was 35.0% by IIF, 41.6% by CENP-A and 57.8% by CENP-B ELISA. When the CENP-A and the CENP-B ELISA results were compared to the IIF, the area under the curve value derived from receiver operating characteristic analysis was 0.98 for both assays. ACA and anti-topoisomerase I antibodies co-occurred in 1.2% (ACA by IIF), in 3.5% (by CENP-A ELISA) and in 7.4% (by CENP-B ELISA). Anti-CENP-A antibodies were negatively associated with anti-Scl-70, anti-RNA Pol III, (both p<0.0001), anti-U1-RNP (p=0.008) and anti-PM1-Alpha antibodies (p=0.0337). The degree of association was dependent on the cut-off value used. Although we found good agreement between IIF and ELISA for the detection of ACA in SSc, a significant portion of CENP ELISA positive sera did not show the typical ACA staining pattern. Based on these findings, we conclude that an IIF ACA negative result might not rule out the presence of ACA. In addition, new CENP ELISA kits are reliable for the detection of anti-CENP in SSc sera.

Mahler M ，You D ，Baron M ，Taillefer SS ，Hudson M ，Canadian Scleroderma Research Group(CSRG) ，Fritzler MJ ... -  《-》