摘要
背景以往研究证明葡萄膜炎的发病机制与γδT细胞相关，18T细胞在实验性自身免疫性葡萄膜炎（EAU）中的作用尚不完全清楚。目的研究γδT细胞在EAU小鼠脾脏中的动态变化，探讨γδT细胞在EAU病程进展中的作用机制。方法应用随机数字表法将45只C57BL／6（B6）小鼠随机分为正常对照组6只和模型组39只。用抗原肽光感受器间维生素A类结合蛋白1-20（IRBP1-20）及完全弗氏佐剂（CFA）的乳化液在B6小鼠的足垫、尾根部及躯干部均匀注射6个点，用Genesis-D动物眼部照相机观察并记录正常小鼠及免疫后不同时间点（4、8、12、16、20、24、28、32和36d）EAU小鼠的发病情况，参照Thurau的评分标准进行炎症评分。颈椎脱臼法处死小鼠后摘取右侧眼球，切片后行组织病理学评分。同时在相同时间点分离模型小鼠脾脏淋巴细胞，流式细胞仪检测18T细胞数量和激活状态。免疫磁珠分选18T细胞，并用流式细胞仪检测其细胞内表达白细胞介素-17A（IL-17A）的变化。向EAU小鼠回输激活的18T细胞，观察炎症变化。结果经IRBP1-20及CFA乳化液免疫后小鼠于第12天可见轻度葡萄膜炎症，炎症反应在免疫后第16-20d达峰，至第28天炎症明显减轻。组织病理学观察发现，免疫小鼠视网膜外核层皱褶，玻璃体、视网膜全层炎性细胞浸润及内界膜层增厚。造模后不同时间点的炎症症状评分和病理学炎症评分的总体比较，差异均有统计学意义（F=51．399，P=0．000；F=47．342，P=0．000）。流式细胞仪检测发现，EAU发病高峰期小鼠的脾脏中γδT细胞数量增加，造模后16d和20d分别为（5．67±0．49）％和（5．78±0．55）％，与造模前的（1．53±0．14）％比较，差异均有统计学意义（均P〈0．05），且呈活化状态。EAU发病高峰期小鼠的脾脏中18T细胞分泌的IL-17A明显增多，造模后16d和20d分别为（13．40±0．50）％�
Background Previous studies showed that the pathogenesis of uveitis is related to γδ T cells. However,it remains unclear that how these cells are involved in experimental autoimmune uveitis （EAU）. Objective This study aimed to observe the dynamic changes of γδ T cells in EAU and explore the role of γδ T cells in the pathological process of EAU. Methods Forty-five C57BL/6 （B6） mice were assigned to the normal control group （six mice） and EAU model group （thirty-nine mice）. The mice were immunized subcutaneously at 6 spots on the footpads, tail base, and flank with emulsion containing human interphotoreceptor retinoid binding protein1-20 （IRBP1-20） emulsified in complete Freund＇s adjuvant. After immunization,the mice were examined for clinical signs of EAU by using a Genesis-D camera. The changes of histopathology were compared by hematoxylin and eosin staining. Mouse lymphocytes were isolated and purified from the spleens of IRBP1-20-immunized or normal B6 mice by using a γδ T-cell isolation kit. Flow cytometry was used to detect the changes of intracellular expression of interleukin- 17A （IL-17A） , and then transferred the activated γδ T cells into EAU models to analyze the changes of clinical signs and histopathology of EAU. Experimental study program as well as the use and feeding of the animals were authorized by the Animal Management and Use Committee of Shandong Traditional Chinese Medicine University. Results The inflammatory symptoms in mouse eyes appeared on day 12 after modeling. The initial changes were fundal blood vessel thickening and minimal inflammatory cell infiltration. Then, muhifocal chorioretinal lesions, serious vasculitis and linear lesions were observed on days 16-20,along with abundant lymphocyte infiltration in the vitreous and retinal disorganization. The inflammation symptom scores and the pathological inflammation scores at different time points after modeling had statistically significant differences （ F = 51. 399, P = 0. 000; F = 47. 342, P = 0.
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2017年第9期793-798,共6页
Chinese Journal Of Experimental Ophthalmology
基金
国家自然科学基金项目（81373826）
