Proteomic surveillance of autoantigens in relapsing polychondritis.
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 cartilagerelated 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.
Institute of Medical Science, St. Marianna University School of Medicine, Kanagawa, Japan.
Yasuhiko
Y
Tanaka
Manabu
M
Nakamura
Toshihiro
T
Matsui
Nobuko
N
Iizuka
Hirobumi
H
Kondo
Shigeto
S
Tohma
Kayo
K
Masuko
Kazuo
K
Yudoh
Hiroshi
H
Nakamura
Kusuki
K
Nishioka
Izumi
I
Koizuka
Tomohiro
T
Kato
D001323:Autoantibodies / Q000097:blood; D001324:Autoantigens / Q000276:immunology* / Q000302:isolation & purification; D014408:Biomarkers, Tumor / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D037282:Calreticulin / Q000276:immunology / Q000302:isolation & purification; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000276:immunology / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins; D006801:Humans; D008297:Male; D008875:Middle Aged; D010751:Phosphopyruvate Hydratase / Q000276:immunology / Q000302:isolation & purification; D011081:Polychondritis, Relapsing / Q000276:immunology*; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000097:blood / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014404:Tubulin / Q000032:analysis / Q000276:immunology; D025521:Tumor Suppressor Proteins / Q000276:immunology / Q000302:isolation & purification; D014746:Vimentin / Q000276:immunology / Q000302:isolation & purification
D001323:Autoantibodies; D001324:Autoantigens; D014408:Biomarkers, Tumor; D037282:Calreticulin; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D011994:Recombinant Proteins; D014404:Tubulin; D025521:Tumor Suppressor Proteins; D014746:Vimentin; C046289:colligin; C095157:ENO1 protein, human; D010751:Phosphopyruvate Hydratase