Proteomic surveillance of retinal autoantigens in endogenous uveitis: implication of esterase D and brain-type creatine kinase as novel autoantigens
Proteomic surveillance of retinal autoantigens in endogenous uveitis: implication of esterase D and brain-type creatine kinase as novel autoantigens
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发表时间：
2008-06
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影响因子：
2.2
通讯作者：
Y. Okunuki;Y. Usui;T. Kezuka;T. Hattori;K. Masuko;Hiroshi Nakamura;K. Yudoh;H. Goto;M. Usui;K. Nishioka;Tomohiro Kato;M. Takeuchi
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作者：
Y. Okunuki;Y. Usui;T. Kezuka;T. Hattori;K. Masuko;Hiroshi Nakamura;K. Yudoh;H. Goto;M. Usui;K. Nishioka;Tomohiro Kato;M. Takeuchi
目的在组织破坏性的内源性葡萄膜炎过程中，各种视网膜蛋白质被新暴露于免疫系统。这些蛋白质中的一些可能是自身抗原，其延长了人类内源性葡萄膜炎中的眼部炎症。在这项研究中，我们的目的是调查的可能性，这种传播的自身抗原在内源性葡萄膜视网膜炎使用蛋白质组学的方法。方法用视网膜光感受器间维甲酸结合蛋白（IRBP）1-20位氨基酸组成的多肽（GPTHLLP PSLVLDMAKVLLP）诱导小鼠实验性自身免疫性葡萄膜视网膜炎（EAU）。免疫后6周，EAU小鼠视网膜蛋白的自身抗体的存在进行了检查，通过双向电泳，然后通过蛋白质印迹（2D-WB）。通过质谱（MS）鉴定由自身抗体靶向的视网膜蛋白，并通过酶联免疫吸附试验检测内源性葡萄膜炎患者（例如白塞病（BD，n=36）、Vogt-小柳-原田病（VKH，n=16）和结节病（n=17））的自身抗原性。结果2D-WD在EAU小鼠中检测到6个新的候选自身抗原，经MS鉴定为β-actin、酯酶D（EsteD）、微管蛋白β-2、脑型肌酸激酶（BB-CK）、电压依赖性阴离子选择性通道蛋白和天冬氨酸转氨酶。在内源性葡萄膜炎患者中，25%的BD和25%的VKH患者抗EsteD抗体阳性，25%的VKH和38.4%的结节病患者抗BB-CK抗体阳性。结论EAU诱导的EsteD和BB-CK自身抗体在部分内源性葡萄膜炎患者中也可检测到，提示这些蛋白可能是内源性葡萄膜视网膜炎过程中传播的自身抗原。
Purpose 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 β-actin, esterase D (EsteD), tubulin β-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.