Higher incidences of anti-SS-A/Ro and anti-SS-B/La autoantibodies in Japanese patients with autoimmune disorders —studies of antigens and antibodies using the immunoblotting method

Summary

A specific and sensitive assay was performed to detect both anti-SS-A/Ro, and anti-SS-B/La antibodies in sera of patients with autoimmune disease, including systemic lupus erythematosus, (SLE), progressive systemic sclerosis (PSS), Sjögren's syndrome (SS), discoid lupus erythematosus (DLE), mixed connective tissue disease (MCTD), generalized morphea (GM), and dermatomyositis (DM). The SS-A/Ro and SS-B/La antigens were prepared from human spleen (HSE) and cultured human cell line (KB cells, KBE), while rabbit thymus extract (RTE) was used as the SS-B/La antigen marker. The antigens were partially purified by DEAE cellulose column chromatography. Immunoblotting showed that the SS-A/Ro antibody reacts mainly with the 58-kDa peptide of the partially purified antigen. Sera containing both the SS-A/Ro and SS-B/La antibodies reacted with the 40-kDa peptide of RTE, and the 58-kDA, 42-kDa, and 40-kDa peptides of HSE and KBE. We found that some of the SS-A/Ro antisera could further react with the 64-kDa peptide of HSE and KBE. The 58-kDa peptide is rich in its cytoplasmic fraction of KB cells, and the 40-kDa peptide in nucleoplasmic fraction. The KB cell line is a better source of the antigens than human spleen extract. The immunoblotting method clearly showed that the positivity rates of SS-A/Ro and/or SS-B/La auto-antibodies were higher in sera from Japanese patients with SLE compared with titers reported for Caucasians but not in sera from healthy volunteers.