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目的研究人外周血单个核细胞中HMBPP特异性Vδ2 T细胞细胞因子的产生和表型特征。方法分离人外周血单个核细胞(PBMCs)和Vδ2 T细胞,用不同剂量的HMBPP和抗人CD28联合刺激培养不同的时间。用酶联免疫吸附法和酶联免疫斑点法检测IFN-γ、TNF-α和IL-2的产生,利用流式细胞术检测Vδ2 T细胞中IFN-γ、TNF-α和IL-2的百分比及表型特征。结果 HMBPP以剂量依赖和时间依赖的方式刺激PBMCs产生IFN-γ。流式细胞术从单个细胞水平证实PBMCs中有(2.27±0.50)%CD3+T细胞表达Vδ2,HMBPP特异性刺激Vδ2 T细胞表达IFN-γ[(38.84±2.62)%]和TNF-α[(26.65±2.05)%],其中(19.43±2.10)%Vδ2 T细胞共表达IFN-γ和TNF-α。表型分析显示,HMBPP特异性Vδ2 T细胞主要为效应型和中央型记忆细胞的表型特征,即CD45RO+CCR7+/-CD62L-。HMBPP刺激纯化的γδT细胞获得相同的结果。结论 HMBPP特异性诱导人PBMCs中Vδ2 T细胞产生细胞因子IFN-γ和TNF-α,表现为记忆细胞的表型特征,本研究为探究感染和炎症性疾病中Vδ2 T细胞的功能和免疫学特征提供了依据。
Objective To study the generation and phenotypic characteristics of HMBPP-specific Vδ2 T cell cytokines in human peripheral blood mononuclear cells. Methods Human peripheral blood mononuclear cells (PBMCs) and Vδ2 T cells were isolated and cultured with different doses of HMBPP and anti-human CD28 for different times. The production of IFN-γ, TNF-α and IL-2 were detected by enzyme-linked immunosorbent assay and enzyme-linked immunosorbent assay. The percentage of IFN-γ, TNF-α and IL-2 in Vδ2 T cells was detected by flow cytometry And phenotypic characteristics. Results HMBPP stimulated PBMCs to produce IFN-γ in a dose-dependent and time-dependent manner. (2.27 ± 0.50)% CD3 + T cells expressed Vδ2 in PBMCs by flow cytometry. HMBPP specifically stimulated Vδ2 T cells to express IFN-γ [(38.84 ± 2.62)%] and TNF-α [ 26.65 ± 2.05)%], in which (19.43 ± 2.10)% Vδ2 T cells co-expressed IFN-γ and TNF-α. Phenotypic analysis showed that HMBPP-specific Vδ2 T cells are predominantly phenotypic characteristics of effector and central memory cells, namely CD45RO + CCR7 +/- CD62L-. The same result was obtained with HMBPP stimulation of purified γδT cells. Conclusion HMBPP specifically induces the production of cytokines IFN-γ and TNF-α in Vδ2 T cells in human PBMCs, which is characterized by the phenotypic characteristics of memory cells. In this study, to investigate the functional and immunological characteristics of Vδ2 T cells in infected and inflammatory diseases Provided the basis.