To consider the hypothesis that autoimmune mechanisms may contribute to the pathology of equine joint diseases, 3 autoimmune responses were assayed in sera and synovial fluids. IgM-rheumatoid factor and antibodies to heat shock protein 65 kDa were determined by ELISA; anti-nuclear antibodies were assayed by indirect immunofluorescence to whole cell nuclear components. All parameters showed only modest increases, if any and not in a pattern related to disease, although some statistically significant increases were detected. Group analysis showed significantly elevated synovial fluid IgM-rheumatoid factor (IgM-RF) in horses with OA (P < 0.01), traumatised joints (P < 0.01) and articular fractured bones (P < 0.001). There was no significantly increased IgM-RF in the sera of horses with joint disorders compared to control horses. Significantly raised anti-heat shock protein molecular weight 65 kDa (HSP65) antibodies were only found in the synovial fluids of the horses with traumatised joints (P < 0.001). No correlations were found between the synovial fluid and sera levels of IgM-RF or anti-HSP65 antibodies. Synovial fluid anti-HSP65 antibody and IgM-RF levels positively correlated in the OCD (P < 0.001), fracture (P < 0.01) and synovitis (P < 0.05) groups. As antibodies to HSP65 correlated with IgG concentrations in synovial fluids, it is not possible to draw conclusions on HSP roles in joint disease pathogenesis. No serum anti-nuclear antibodies (ANA) were detected by immunofluorescence using rat liver and a human epithelial cell line (HEp-2) as substrates.(ABSTRACT TRUNCATED AT 250 WORDS)