Proteomic Surveillance of Autoantigens in Relapsing Polychondritis

• Published in: Microbiology and immunology 2006-01, Vol.50 (2), p.117-126
• Main Authors: Tanaka, Yasuhiko, Nakamura, Manabu, Matsui, Toshihiro, Iizuka, Nobuko, Kondo, Hirobumi, Tohma, Shigeto, Masuko, Kayo, Yudoh, Kazuo, Nakamura, Hiroshi, Nishioka, Kusuki, Koizuka, Izumi, Kato, Tomohiro
• Format: Article
• Language: English
• Subjects: Autoantibodies - blood; autoantibody; autoantigen; Autoantigens - immunology; Autoantigens - isolation & purification; Biomarkers, Tumor - immunology; Biomarkers, Tumor - isolation & purification; Blotting, Western; Calreticulin - immunology; Calreticulin - isolation & purification; Carrier Proteins - immunology; Carrier Proteins - isolation & purification; DNA-Binding Proteins - immunology; DNA-Binding Proteins - isolation & purification; Electrophoresis, Gel, Two-Dimensional; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Middle Aged; Phosphopyruvate Hydratase - immunology; Phosphopyruvate Hydratase - isolation & purification; Polychondritis, Relapsing - immunology; proteomics; Proteomics - methods; Recombinant Proteins - blood; Recombinant Proteins - immunology; relapsing polychondritis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tubulin - analysis; Tubulin - immunology; Tumor Suppressor Proteins - immunology; Tumor Suppressor Proteins - isolation & purification; Vimentin - immunology; Vimentin - isolation & purification
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.2006.tb03776.x

Relapsing polychondritis (RP) is a systemic inflammatory disease, in which autoimmunity to cartilage‐related components is thought to be involved in its pathogenesis. However, the autoimmune profile in RP has not been studied fully. We therefore investigated autoantibodies/autoantigens in RP comprehensively, by 2‐dimensional electrophoresis (2DE), subsequent western blotting (WB) and mass spectrometry, using cell‐extracted proteins as the antigen source. As a result, we detected 15 autoantigens on 2DE‐WB, and further identified five of them. On average, one RP serum recognized approximately 8 out of the 15 autoantigens. Frequencies of the autoantibodies to the 5 identified antigens of tubulin alpha ubiquitous/6, vimentin, alpha enolase, calreticulin, and colligin‐1/‐2 were 91%, 46%, 36%, 82%, and 36%, respectively. ELISA using recombinant proteins for them revealed that frequencies of the autoantibodies to tubulin alpha ubiquitous, vimentin, alpha enolase, calreticulin, and colligin‐1 were 36%, 64%, 46%, 27%, and 18%, respectively. Our data demonstrated that the autoimmune reaction was not restricted to cartilage‐related components, rather a variety of autoimmune responses occurred in patients with RP, which may be involved in the pathophysiology of RP. In addition, the proteomic approach using cell‐extracted proteins would be a powerful way to investigate autoantigens.