【摘要】 目的初步探讨嗜乳脂蛋白3A1(BTN3A1)在结核杆菌耐热抗原(MTB-HAg)诱导的人外周血Vγ9Vδ2T细胞活化增殖中的作用。方法人外周血单个核细胞(PBMC)先加BTN3A1阻断性抗体作用3 h,再分别用MTB-HAg和磷酸类抗原(PAg)刺激培养24h后收集细胞,用流式细胞术检测Vγ9Vδ2 T细胞中CD69表达;或刺激20 h后收集细胞,检测Vγ9Vδ2 T细胞中1型辅助性T(Th1)细胞因子γ干扰素(IFN-γ)和肿瘤坏死因子α(TNF-α)产生细胞比例;或刺激后,在含白细胞介素2(IL-2)培养液中培养10 d后,收集细胞检测Vγ9Vδ2 T细胞扩增比例及增殖活性。结果 MTB-HAg刺激后,Vγ9Vδ2 T细胞中CD69平均荧光强度和阳性细胞比例在BTN3A1阻断组均明显下降(为刺激组的13.84%和43.00%);而PAg阻断组的CD69平均荧光强度和阳性细胞比例显著被抑制(为刺激组的3.10%, 4.47%和9.53%, 10.91%)。MTB-HAg刺激后Vγ9Vδ2T细胞中IFN-γ和TNF-α产生细胞比例在BTN3A1阻断组显著减少, Vγ9Vδ2T细胞扩增数量和细胞增殖活性在BTN3A1阻断组也显著减少或降低。与PAg阻断组明显被抑制的结果相似。结论 BTN3A1促进MTB-HAg诱导的外周血Vγ9Vδ2T细胞活化增殖。更多还原

【Abstract】 Objective To investigate the role of butyrophilin 3A1(BTN3A1) in the activation and proliferation of human peripheral blood Vγ9Vδ2 T cells induced by M. tuberculosis heat resistant antigen(MTB-HAg). Methods Human peripheral blood mononuclear cells(PBMCs) were treated with BTN3A1 blocking antibody for 3 hours and then stimulated with MTB-HAg or phosphoantigen(PAg). At 24 hours of stimulation, the cells were collected to detect the expression of CD69 in Vγ9Vδ2 T cells by flow cytometry. At 20 hours of stimulation, the cells were collected to detect the proportions of cells producing helper T cell type I(Th1) cytokines IFN-γ and tumor necrosis factor α(TNF-α) in the Vγ9Vδ2 T cells. The PBMCs were also stimulated and cultured in IL-2-containing medium for 10 days, and the expansion and proliferation activity of Vγ9Vδ2 T cells were detected. Results After stimulated with MTB-HAg, the average fluorescence intensity of CD69 and the proportion of CD69 positive cells in Vγ9Vδ2 T cells decreased significantly in BTN3A1 blocked group, being 13.84% and 43.00% of those in the stimulated group, respectively. However, the average fluorescence intensity of CD69 molecules and the proportion of positive cells in PAg blocked group were significantly inhibited(3.10%, 4.47% and 9.53%, 10.91% of those in the stimulated group). The proportions of IFN-γ and TNF-α producing Vγ9Vδ2 T cells stimulated with MTB-HAg decreased significantly in the BTN3A1 blocked group, and the expansion number and cell proliferation activity of Vγ9Vδ2 T cells were also reduced significantly in the BTN3A1 blocked group. The results were similar to those of the PAg blocked group. Conclusion BTN3A1 promotes activation and proliferation of peripheral blood Vγ9Vδ2 T cells induced by MTB-HAg.更多还原