Anti-Ro antibodies are found in a large proportion of patients with systemic lupus erythematosus and primary Sjögren's syndrome. These antibodies also characterize neonatal lupus, subacute cutaneous lupus erythematosus, and vasculitis associated with Sjögren's syndrome and rheumatoid arthritis. Anti-Ro-positive sera may contain either or both of 2 sets of antibodies, recognizing either a 60-kd or a 52-kd polypeptide component of the Ro particle. We found in this study that the immune response to the 60-kd Ro antigen is heterogeneous. Some sera specifically recognize the native Ro antigen but fail to bind the corresponding denatured polypeptides. In addition, after immunodepletion using the denatured 60-kd Ro polypeptide, all anti-Ro-positive sera tested still contained high titers of antibodies recognizing conformational determinants on the Ro antigen. The frequent immunodominance of anti-Ro antibodies targeted to conformational determinants suggests that native autoantigens may directly drive the autoimmune response.