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目的检测卡介苗(BCG)刺激后,PPD+正常人外周血中分泌IFN-γ的TCRγδ亚群及其表型特征。方法分离PPD+正常人外周血单个核细胞(PBMCs),用多色流式细胞术检测TCRγδ亚群;BCG刺激PBMCs后检测γδT细胞IFN-γ分泌水平,并分析BCG特异性γδT细胞表型特征。结果依据PBMCs中TCRγδ和δ2的表达,可将γδT细胞分为3个亚群,即δ2high、δ2low和TCRγδ中非δ2T细胞(non-δ2-γδT)细胞。其中,δ2high和δ2lowT细胞几乎全部(>98%)为CD45RO+,而non-δ2-γδT细胞中只有少部分表达CD45RO,其余大部分不表达CD45RO。δ2high和δ2lowT细胞表型类似,主要为CD45RO+CD62L-CD27-,而non-δ2CD45RO+细胞主要为CD62L-CD27+,non-δ2CD45RO-细胞几乎全部为CD62L-CD27-。BCG刺激PBMCs后,可诱导γδT细胞分泌IFN-γ,且主要为δ2high和δ2low细胞,而non-δ2-γδT几乎不产生IFN-γ。进一步分析BCG特异性γδT细胞的表型特征,结果表明,δ2high-CD45RO+IFN-γ+和δ2lowCD45RO+IFN-γ+细胞主要为CD62L-CD27-和CD62L-CD27+,为效应型记忆细胞。结论 BCG刺激PPD+正常人外周血PBMCs后,主要诱导δ2T细胞分泌IFN-γ,且该细胞表现出CD45RO+CD62L-效应型记忆细胞特征,可能在预防结核早期感染中发挥重要作用。
Objective To detect the TCRγδ subunit of IFN-γ secreted by peripheral blood of PPD + normal people after BCG stimulation and its phenotypic characteristics. Methods Peripheral blood mononuclear cells (PBMCs) from PPD + normal individuals were isolated and TCRγδ subgroups were detected by multi-color flow cytometry. IFN-γ secretion of γδT cells was detected by BCG stimulation, and the phenotypic characteristics of BCG-specific γδT cells were analyzed. Results According to the expression of TCRγδ and δ2 in PBMCs, γδT cells could be divided into three subgroups: δ2high, δ2low and TCRγδ non-δ2T cells (non-δ2-γδT) cells. Among them, almost all (> 98%) of δ2high and δ2low T cells were CD45RO +, while only a small portion of non-δ2-γδT cells expressed CD45RO, while most of the remaining did not express CD45RO. The phenotypes of δ2high and δ2lowT cells were similar, mainly CD45RO + CD62L-CD27-, whereas non-δ2CD45RO + cells were predominantly CD62L-CD27 + and non-δ2CD45RO- cells were almost exclusively CD62L-CD27-. After BCG stimulated PBMCs, γδT cells were induced to secrete IFN-γ, mainly δ2high and δ2low cells, whereas non-δ2-γδT produced almost no IFN-γ. Further analysis of the phenotypic characteristics of BCG-specific γδT cells showed that δ2high-CD45RO + IFN-γ + and δ2lowCD45RO + IFN-γ + cells were predominantly CD62L-CD27- and CD62L-CD27 + and were effector memory cells. CONCLUSION: After BCG stimulates peripheral blood PBMCs from PPD + normal people, IFN-γ is mainly induced by δ2T cells, and the cells show characteristics of CD45RO + CD62L-effector memory cells, which may play an important role in preventing early TB infection.