荧光染料CFSE在实验性葡萄膜炎T细胞增殖研究中的应用

Tracking T Lymphocytes Proliferation with Vital Dye CFSE in Experimental Autoimmune Uveitis

目的探讨荧光染料CFSE在实验性葡萄膜炎(EAU)抗原特异T细胞增殖研究中的应用价值。方法以人类光感受器间维生素A类结合蛋白(IRBP)的多肽片段IRBP161-180为抗原,免疫B10RⅢ小鼠使其产生EAU。用CFSE荧光染料标记细胞,结合荧光单抗和流式细胞术,检测T细胞及其亚群在特异抗原刺激下,不同时间点细胞增殖的情况。结果IRBP161-180刺激4 d后出现T淋巴细胞增殖分裂,表现为CFSE荧光强度的系列减半。使用荧光单抗双标记发现CD4+T和CD8+T细胞增殖反应不同步,CD8+T细胞较CD4+T细胞增殖分裂更活跃,亲代细胞百分率下降更明显(P<0.01)。结论CFSE荧光标记技术是分析EAU抗原特异性T细胞及亚群增殖分裂的有力工具。

Objective To track T lymphocytes proliferation with vital dye CFSE in experimental autoimmune uveitis（EAU）.Methods B10RIII mice were immunized with synthetic peptide of interphotoreceptor retinal-binding protein（IRBP161-180） to develop EAU.The proliferation of IRBP-specific T cell subsets was detected by CFSE staining,fluorescent antibody labeling,and flow cytometry.Results The IRBP-specific T cells divided after 4 days of stimulation with IRBP161-180,halving the fluorescence intensity.The proliferations of CD4 ＋ and CD8 ＋ T cells were asynchronous,with CD8 ＋ T cells dividing more vigorously and having more drop in percentage of parent cells（P〈0.01）.Conclusion CFSE-Labeling can detect the proliferation of autoreactive T cells and their subsets in EAU effectively.