Using 2-dimensional electrophoresis and ion-pair chromatography, we have identified elements of
proliferating cell nuclear antigen (
PCNA)
multiprotein complexes that are reactive to
antibodies in sera from patients with
systemic lupus erythematosus. Among the various elements of the complexes, a 37 kDa
protein (PI 8.5) that specifically reacted with SLE sera, but not with sera from patients with other
connective tissue diseases, was identified as
glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Immunoblot analysis showed that SLE sera reactive with the 37 kDa
protein specifically reacted with GAPDH, as did anti-GAPDH mAbs. The purified
autoantibodies to GAPDH from lupus serum showed both nuclear speckled and cytoplasmic staining patterns in immunofluorescence on Hep-2 cells. In addition,
enzyme-linked
immunosorbent assay (ELISA) revealed the presence of anti-GAPDH
autoantibodies in 47% of lupus patients. Longitudinal analysis of the reactivity of lupus sera to
PCNA complexes showed the autoimmune response to spread from GAPDH to other elements of
PCNA complexes, and the presence of anti-GAPDH
antibodies was significantly correlated with increased levels of serum
PCNA. Taken together, these findings suggest that GAPDH interacting with
PCNA in association with its cellular function is a novel
autoantigen recognized by lupus sera, and that GAPDH thus plays an important role in the induction of autoimmune responses against the
PCNA complex.