Volume 3 Supplement 2
21st European Workshop for Rheumatology Research
Expression of citrullin-containing antigens in RA synovium
© BioMed Central Ltd 2001
Received: 15 January 2001
Published: 26 January 2001
The presence of autoantibodies directed to citrulli-nated antigens in serum is highly specific for RA. The aim of this study was to compare anti-CCP concentrations in paired serum/SF samples of patients with RA and to investigate whether this is associated with the expression of citrullinated antigens in RA synovium and to study the nature of these antigens.
A recombinant single-chain variable fragment (scFv) mon-oclonal antibody was selected against a cyclic citrullinated peptide (CCP) from a patient antibody-fragment phage-display library. This scFv and patient antibodies affinity purified with CCP, were both used for immunohistochemical staining of synovial cryostat sections of RA (30) and control patients (OA (13), ReA (9), and other arthri-tides (28)). In addition, rabbit anti-citrullin antibodies (Biogenesis) were used for immunohistochemistry of synovial cryostat sections of RA (14), and control patients (OA (10), ReA (7), and other arthri-tides (23)). IgG anti CCP titers were calculated with the quantitative Rapscan RA ELISA kit (Eurodiagnostica). Total IgG concentrations were determined on a cobas Fara-2 centrifugal analyzer.
Citrullin containing antigens were observed in synovial cryostat sections of anti-CCP positive and negative patients. Staining with ScFv monoclonal antibody was noted in synovial lining cells and in (peri)vascular areas in 13/30 RA patients, 7/13 OA patients, 5/9 ReA patients, and 12/28 other arthritis patients. CCP positivity was on average similar in all diagnostic groups. Staining was absent in the negative controls using a control scFv antibody. Staining with rabbit anti-citrullin polyclonal antibody was noted in 8/14 RA patients, 3/10 OA patients, 2/7 ReA patients, and 6/23 other arthri-tides. However, controls using irrelevant rabbit antibodies were also positive in some patients in all groups. Anti-CCP concentrations (expressed in Units per mg total IgG) were on average 1.34 times higher in SF compared to serum (n = 20, P < 0.05) or 1.37 when only positive samples were included (n = 11, P < 0.05)
Citrullinated antigens are present in the synovia of both RA and control patients with similar prevalence. The presence of anti-CCP autoantibodies in serum is not associated with the expression of citrullinated antigens in the synovium.
The identity of the citrullinated antigens and potential differences between RA and control synovia remain to be identified.