八色流式细胞术检测人外周血BCG特异性效应型记忆CD4^+ T细胞的表型特征

Phenotypic analysis of BCG-specific effector memory CD4^+ T cells in PBMCs by eight-color flow cytometry

目的:检测BCG刺激后,PPD+正常人外周血中细胞因子产生及其亚群。方法:分离PPD+正常人外周血单个核细胞(PBMC),BCG刺激后检测CD4+和CD8+ T细胞细胞因子分泌,并用八色流式细胞术分析BCG特异性T细胞亚群。结果:BCG刺激PBMC后,主要是CD4+T细胞分泌Th1细胞因子(IFN-γ、IL-2和TNF-α),而CD8+T细胞几乎不产生细胞因子。进一步分析分泌细胞因子的细胞亚群,主要是CD4+CD45RO+CD62L-CD27-和CD4+CD45RO+CD62L-CD27+分泌细胞因子。结论:BCG刺激PPD+正常人外周血PBMC后,主要诱导CD4+ T细胞分泌细胞因子,且该细胞表现出CD4+CD45RO+CD62L-效应型记忆细胞特征,可能在预防结核感染中发挥重要作用。

AIM： To evaluate cytokine production and subsets in PBMCs from PPD＋ normal donors after stimulation with BCG.METHODS： PBMCs were isolated from PPD＋ normal individuals,and cytokine production and BCG-specific T cell subsets were assessed by eight-color flow cytometry.RESULTS： Following stimulation with BCG,CD4＋ but not CD8＋ T cells expressed IFN-γ,IL-2 and TNF-α.Phenotypic analysis indicated that cytokine-producing cells were CD4＋CD45RO＋CD62L-CD27-and CD4＋CD45RO＋CD62L-CD27＋.CONCLUSION： BCG predominantly induced CD4＋ T cells to produce cytokines following stimulation with BCG.Further analysis indicated that these cells are CD4＋CD45RO＋CD62L-effector memory cells,suggesting that these cells probably played essential role in preventing TB infection.