Specificity of autoantibodies to SS-A/Ro on a transfected and overexpressed human 60 kDa Ro autoantigen substrate.

The objective of this study was to analyze apparently discrepant results that arose during the use of an indirect immunofluorescence (IIF) assay using transfected HEp-2 cells to detect anti-SS-A/Ro autoantibodies in human sera. Fourteen sera that had SS-A/Ro antibodies as detected on this commercial substrate, but did not have antibodies to SS-A/Ro as determined by double immunodiffusion (ID) or enzyme-linked immunosorbent assay (ELISA), were studied by immunoprecipitation (IP) of radiolabeled cell extracts and full-length recombinant SS-A/Ro. A multi-antigen strip immunoblotting (IB) assay containing both the 52- and 60-kDa antigens was included in the analysis. We confirmed that 12 of 14 of the sera under study had antibodies to SS-A/Ro protein antigens as determined by at least one other immunoassay. One serum had antibodies to hyRNA but no detectable reactivity with the 52- or 60-kDa antigens. One serum remained negative in all assays for SS-A/Ro autoantibodies.