嗜乳脂蛋白3A1(BTN3A1)促进结核杆菌耐热抗原诱导的人外周血Vγ9Vδ2 T细胞活化增殖  

Butyrophilin 3A1(BTN3A1)enhances activation and proliferation of human peripheral blood Vγ9Vδ2 T cells induced by MTB-HAg

作　　者：唐洁[1,2,3] 孙静[3] 查成 常见荣 胡坤[3] 方强[2] 陈雨晴 陈静 李柏青[1,2] TANG Jie;SUN Jing;ZHA Cheng;CHANG Jianrong;HU Kun;FANG Qiang;CHEN Yuqing;CHEN Jing;LI Baiqing(Department of Immunology,Bengbu Medical College,Bengbu 233000,China;Anhui Key Laboratory of Infection and Immunity,Bengbu Medical College,Bengbu 233000,China;Science Research Centre,Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院免疫学教研室,安徽蚌埠233000 [2]安徽省感染与免疫重点实验室(蚌埠医学院),安徽蚌埠233000 [3]蚌埠医学院科研中心,安徽蚌埠233000

出　　处：《细胞与分子免疫学杂志》2020年第8期680-686,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：安徽省高校自然科学研究重点项目(KJ2018A0227);安徽高校科研平台创新团队项目(2016-40);蚌埠医学院自然科学基金重点项目(BYKY1813ZD)。

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Vγ9Vδ2 T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24 h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Vγ9Vδ2 T细胞中IFN-γ和TNF-α产生细胞比例在BTN3A1阻断组显著减少,Vγ9Vδ2 T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Vγ9Vδ2 T细胞活化增殖。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-γand tumor necrosis factorα(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-γand TNF-αproducing 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.

关 键 词：嗜乳脂蛋白3A1(BTN3A1) 结核杆菌耐热抗原(MTB-HAg) 磷酸类抗原(PAg) Vγ9Vδ2 T细胞 

分 类 号：R378.911[医药卫生—病原生物学] R392.1[医药卫生—基础医学]