The aim of the present study was to detect antigenic proteins that react specifically with cerebrospinal fluid (CSF)-IgG from oligoclonal IgG bands (OB)-positive multiple sclerosis (MS) patients. To identify such antigenic proteins, we developed a rat brain proteome map using 2-DE and applied it to the immunoscreening of brain proteins that react with CSF-IgG but not with serum-IgG in OB-positive MS patients. After sequential MALDI-TOF mass spectrometry, eight proteins [two neuronal proteins (tubulin β-2 and γ enolase-2), HSP-1, Tpi-1 protein and cellular enzymes (creatine kinase, phosphopyruvate hydratase, triosephosphate isomerase and phosphoglycerate kinase-1)] were identified as candidate antigens in seven MS patients. Reactivity to tubulin was seen in Western blotting in four patients, and CSF-specific anti-tubulin IgG was detected in one patient. In addition, CSF-specific anti-gamma enolase IgG was found in another patient. These findings suggest that intrathecal immune responses may occur against a broad range of proteins in MS.
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