抗肿瘤坏死因子-α单克隆抗体治疗实验性自身免疫性葡萄膜视网膜炎的研究

Treatment of experimental autoimmune uveoretinitis with anti-twnor necrosis factor-α monoclonal antibody

目的 观察抗肿瘤坏死因子-α单克隆抗体（TNF—αMCAb）对于实验性自身免疫性葡萄膜视网膜炎（EAU）的治疗作用。方法合成光感受器间维生素A类结合蛋白（IRBP）R16多肽片段，联合免疫佐剂诱导EAU动物模型。按照注射次数不同将大鼠分为2组，分别于IRBPR16免疫后的第6天和第4、6、8天自大鼠尾静脉注入TNF—α MCAb，裂隙灯显微镜观察其眼部临床表现，同时设未治疗组大鼠作为对照。于IRBPR16免疫后第13天测量迟发型超敏反应（DTH），并于第14天处死大鼠，取眼球行组织病理检查。应用酶联免疫吸附试验（ELISA）检测抗体注射后14d时大鼠血清中Th1类细胞因子γ-干扰素（IFN-γ）、Th2类细胞因子白细胞介素-4（IL-4）水平以及房水中IFN-γ水平。测量引流淋巴结细胞的抗原特异性淋巴细胞增生反应。结果TNF—α MCAb治疗组大鼠的眼部炎症较未治疗组明显减轻，病理分级下降；房水和血清中IFN-γ水平降低，血清中IL-4增高；DTH反应下降；引流淋巴结细胞对IRBPR16多肽刺激的增生反应下降。差异均有统计学意义（P〈0．01）。与单次注射相比，3次注射的治疗效果更显著（P〈0．05）。结论TNF-αMCAb能够有效减轻EAU大鼠眼部的炎症反应和特异性细胞免疫反应，改变Th1／Th2细胞因子平衡；多次应用作用更显著。

Objective To observe the efficacy of the anti-tumor necrosis factor-α monoclonal antibody （TNF-α MCAb） in the treatment of experimental autoimmune uveoretinitis （EAU）. Methods EAU animal models were induced by interphotoreceptor retinoid-binding protein （IRBP） R16 peptide with immunization. The rats were divided into 2 groups according to the injection times. TNF-α MCAb was administered intravenously on day 6 or 4, 6 and 8 post-immunization respectively, and then to observe the clinical expression by slit lamp microscope. Meanwhile, take the rats which did not accept TNF-α MCAb as control group. Delayed type hypersensitivity （DTH） responses were measured on day 13 post- immunization of IRBP R16 the rats were killed on day 14 post-immunization of IRBP R16, and then enucleated the eyes for histopathological examination. To detect the cytokine level of IFN-γ, IL-4 in serum and IFN-γ in aqueous humor by enzyme liked immunosorbent assay （ELISA） on day 14 postinjection. The hyperplasia responses of antigen specific lymphocyte of draining lymph node ceils were detected. Results The TNF-α MCAb group had mitigated ocular inflammation and decreased pathological grades compared with the control group; the IFN-γ concentrations in aqueous humor and serum were decreased, IL-4 was increased in serum; DTH responses were decreased; the hyperplasia responses of draining lymphocytes to IRBP R16 peptide were decreased, all the differences were statistically significant （P〈0. 01）. The rats accepted TNF-a MCAb thrice had much better curative effect than the rats injected once （P 〈0.05）. Conclusions Injection of TNF-α MCAb can inhibit ocular inflammation and specific immune cells of EAU remarkably and change the Th1/Th2 balance. Many times injections of TNF-α MCAb were more effective than once.