Proteomic surveillance of retinal autoantigens in endogenous uveitis: implication of esterase D and brain-type creatine kinase as novel autoantigens.
OBJECTIVE
Various retinal proteins are newly exposed to immune system in a process of tissue destructive endogenous uveitis. Some of such proteins could be autoantigens that extend the ocular inflammation in human endogenous uveitis. In this study, we aimed to investigate the possibility of such spreading of autoantigens in endogenous uveoretinitis using a proteomic approach.


METHODS
Experimental autoimmune uveoretinitis (EAU) was induced in mice by inoculation with a peptide consisting of amino acids 1-20 (GPTHLFQPSLVLDMAKVLLP) of interphotoreceptor retinoid binding protein (IRBP). Six weeks after immunization, the presence of autoantibodies against the retinal proteins in mice with EAU were examined by two-dimensional electrophoresis followed by western blotting (2D-WB). Retinal proteins targeted by the autoantibodies were identified by mass spectrometry (MS) and their autoantigenicity in patients with endogenous uveitis, such as Behcet's disease (BD, n=36), Vogt-Koyanagi-Harada disease (VKH, n=16), and sarcoidosis (n=17) were examined by enzyme-linked immunosorbent assay.


RESULTS
Six new candidate autoantigens, which were detected in mice with EAU using 2D-WD were identified by MS as beta-actin, esterase D (EsteD), tubulin beta-2, brain-type creatine kinase (BB-CK), voltage-dependent anion-selective channel protein, and aspartate aminotransferase. Among the patients with endogenous uveitis, 25% of BD and 25% of VKH patients were positive for anti-EsteD antibody, and 25% of VKH and 38.4% of sarcoidosis patients were positive for anti-BB-CK antibody.


CONCLUSIONS
Autoantibodies to EsteD and BB-CK produced in EAU-induced mice were also detected in some endogenous uveitis patients, suggesting that these proteins might be autoantigens spreading in a process of endogenous uveoretinitis.
Department of Ophthalmology, Tokyo Medical University, Tokyo, Japan.
Yoko
Y
Okunuki
Yoshihiko
Y
Usui
Takeshi
T
Kezuka
Takaaki
T
Hattori
Kayo
K
Masuko
Hiroshi
H
Nakamura
Kazuo
K
Yudoh
Hiroshi
H
Goto
Masahiko
M
Usui
Kusuki
K
Nishioka
Tomohiro
T
Kato
Masaru
M
Takeuchi
D000328:Adult; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001324:Autoantigens / Q000276:immunology*; D001327:Autoimmune Diseases / Q000201:enzymology; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D043182:Carboxylesterase / Q000276:immunology*; D003402:Creatine Kinase / Q000276:immunology*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007074:Immunoglobulin G; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D040901:Proteomics; D012160:Retina / Q000201:enzymology / Q000276:immunology*; D014605:Uveitis / Q000201:enzymology* / Q000276:immunology*
D000906:Antibodies; D001324:Autoantigens; D007074:Immunoglobulin G; D003402:Creatine Kinase; D043182:Carboxylesterase; C020118:ESD protein, human