正常人外周血Th17细胞的频率、表型及其功能特征

摘要

目的:研究正常人外周血Th17细胞的频率、表型及其功能特征.方法:分离正常人外周血单个核细胞(PBMCs),加入刺激剂PMA和离子霉素,同时加入蛋白转运抑制剂BFA,细胞置于37℃、50 mL/LCO2培养箱培养4～5h.收集细胞,以荧光素-mAb标记后,用流式细胞仪检测.结果:正常人PBMCs经PMA+ Ionomycin刺激后,Th17细胞的频率为(1.63±0.72)％,细胞表达CD45RO (99.32±0.79)％、CCR6(97.37±3.04)％和CCR4 (96.27±3.49)％；约15％的Th17细胞表达IFN-γ,超过80％的Th17细胞表达IL-2,但Th17细胞几乎不表达IL-10或IL-4.结论:正常人外周血中存在记忆性Th17细胞,该细胞表达趋化因子受体CCR6和CCR4,且具有不同于Th1和Th2细胞的功能特征.%Objective To analyze the frequency,phenotype and functional characterization of peripheral blood Th17 cells in healthy individuals.Methods PBMCs were isolated from normal health human peripheral blood and cultured with PMA and ionomycin in the presence of Brefeldin A for 4 ～ 5 h at 37 ℃ in a 5％ CO2humidified atmosphere.The frequency of subpopulation of IL-17-producing cells,the expression of IL-17 with Th1 and Th2 cytokines and the phenotypic characterization of IL-17-producing cells were assessed by flow cytometry.Results The frequency of Th17 cells was (1.63 ± 0.72)％ following stimulation with PMA plus ionomycin.Th17 cells express CD45RO,CCR6 and CCR4.About fifteen percentage of Th17 cells showed a double IL-17+ IFN-γ+ phenotype,more than eighty percentage of Th17 cells showed a double IL-17+IL-2+phenotype,whereas cells producing both IL-17 and Th2 cytokines were almost undetectable.Conclusion Memory Th17 cells with the expression of CCR6 and CCR4 are apparent in healthy individual PBMCs and Th17 cells are different from Th1/Th2 cells.