IRBPR16多肽的合成及其致葡萄膜视网膜炎活性的探讨

目的 探讨光感受器间维生素A类结合蛋白（IRBP）的R16多肽片段的致葡萄膜视网膜炎活性。设计实验研究。研究对象36只Lewis大鼠。方法应用Fmoc法合成并纯化牛IRBPR16多肽片段，以诱导实验性自身免疫性葡萄膜视网膜炎（EAU）模型，并对该模型进行临床观察和组织学检查。培养EAU大鼠的引流淋巴结细胞，测定淋巴细胞增殖反应。各实验同时建立单纯弗式完全佐剂（CFA）免疫组和空白对照组。主要指标多肽分析，视网膜形态学，淋巴细胞增殖反应。结果合成的IRBPR16多肽片段纯度为95．6％。应用IRBPR16多肽片段作为抗原免疫Lewis大鼠，可成功诱导出EAU模型。EAU的临床分级为（3．33±0．52）级，病理分级为（3．67±0．92）级；CFA组和空白对照组大鼠眼部均无异常改变。EAU组大鼠引流淋巴结中抗原特异性淋巴细胞增殖反应增强，为（33．27±7．24）×10^cpm，显著高于CFA组（1．91±1．16）×10^3cpm]和空白对照组（1．23±0．51）×10^3cpm]（P〈0．05）。结论IRBPR16多肽片段具有较强的致葡萄膜视网膜炎活性，引流淋巴结抗原特异性淋巴细胞增殖反应增强。IRBPR16多肽诱导的EAU为研究人类葡萄膜视网膜炎提供了一个重要的动物模型。

Synthesis of IRBP R16 peptide and its uveitogenic activity

Objective To investigate the uveitogenesity of R16 peptide from bovine interphotoreceptor retinoid-binding protein （IRBP）. Design Experimental study. Participates 36 Lewis rats. Methods IRBP R16 peptide was synthesized using Fmoc procedure to induce experimental autoimmune uveoretinitis （EAU） model. The clinical scores and pathological grades of EAU were recorded ac- cording to slit-lamp microscopy and histological examination. Lymphoproliferation assay （LPA） was performed using the draining lymph node cells in EAU rats. The Lewis rats received complete Freund＇s adjuvant （CFA） immunization served as CFA controls and the normal ones as negative controls. Main Outcome Measures Peptide analysis, retinal morphology, lymphocyte proliferation response. Results The purity of synthesized IRBP R16 peptide was 95.6 %. EAU model in Lewis rats was successfully induced with IRBP R16 peptide. The clinical scores and pathological grades of EAU were 3.33±0.52 and 3.67＋0.92, respectively. No abnormal signs were observed in either CFA controls or negative controls. LPA showed that the level of 3H-thymidine incorporation of the draining lymph node cells in EAU rats was （33.27± 7.24）×10^3 cpm , which was much higher than that in the CFA group（（ 1.91±1.16）×10^3 epm）and control group （（ 1.23 ×0.51 ）×10^3 cpm）（P〈0.05 ）. Conclusions IRBP R16 peptide is an immunogenic and highly uveitogenic epitope. The response of lymphocyte proliferation in draining lymph node cells is increased in EAU rats induced by R16 peptide. This provides us an important animal model for the study of human uveitis.