3种分枝杆菌耐热蛋白抗原对人γδT细胞的刺激活性研究  

Comparative study on the stimulatory effect of three different heat-treated protein antigens of Mycobacteriumon human γδT lymphocytes

作　　者：陈勇[1] 张涛[1] 刘勇[1] 管俊昌[1] 夏惠[1] 

机构地区：[1]蚌埠医学院病原生物学教研室,感染与免疫安徽省重点实验室,安徽蚌埠233030

出　　处：《中国病原生物学杂志》2016年第11期986-990,共5页Journal of Pathogen Biology

基　　金：感染与免疫安徽省重点实验室(蚌埠医学院)开放课题(No.BYKL1301)

摘　　要：目的研究3种分枝杆菌耐热蛋白抗原体外对人γδT细胞的刺激活性。方法分离健康人外周血单个核细胞(PBMC),分别使用人型结核分枝杆菌H37Ra耐热蛋白抗原(H37Ra-HAg)、牛型结核分枝杆菌BCG耐热蛋白抗原(BCG-HAg)和耻垢分枝杆菌耐热蛋白抗原(MS-HAg)进行刺激并持续培养12d,采用流式细胞仪检测增殖细胞中γδT细胞数量及CD4+T细胞所占比例。结果 H37Ra-HAg刺激组扩增细胞中γδT细胞的比例和数量均显著高于BCGHAg、MS-HAg刺激组和IL-2对照组[((50.52±5.40)%v.s(7.31±3.15)%、(5.82±3.42)%、(5.56±2.68)%);(9.63±1.64)×106v.s(0.69±0.36)×106、(0.57±0.22)×106、(0.34±0.18)×106))]。BCG-HAg刺激组扩增细胞中CD4+T细胞的比例和数量均显著高于H37Ra-HAg刺激组和IL-2对照组[(73.56±7.12%、66.93±7.65)%v.s(23.18±6.29)%、(52.61±5.37)%;(4.75±0.86)×106、(3.28±0.59)×106 v.s(2.71±0.65)×106、(1.63±0.45)×106]。而BCG-HAg刺激组和MS-HAg刺激组扩增细胞中γδT细胞的比例和数量和IL-2对照组相比并无显著性差异。结论 H37Ra-HAg具有促进γδT细胞增殖活性,BCG-HAg和MS-HAg具有促进CD4+T细胞增殖活性,BCG-HAg和MS-HAg无刺激γδT细胞增殖活性。Objective To investigate the stimulatory effect of three different heat-treated protein antigens of Mycobacteriumon humanγδT lymphocytes in vitro. Methods Peripheral blood mononuclear cells（PBMCs）isolated from normal subjects were stimulated with three different heat-treated protein antigens from the H37 Ra strain of Mycobacteriumtuberculosis（H37Ra-HAg）,the BCG strain of M.tuberculosis（BCG-HAg）,and Mycobacteriumsmegmatis（MS-HAg）for12days.The phenotype of expanded lymphocytes was detected with flow cytometry,and the number of expandedγδT cells was counted and the proportion of CD4＋T cells was analyzed. Results The proportion ofγδT cells and the cell count were significantly higher in the H37Ra-HAg group than in the BCG-HAg group,the MS-HAg group,and the IL-2group（50.52±5.40% vs.7.31±3.15%,5.82±3.42%,and 5.56±2.68%;9.63±1.64×106 vs.0.69±0.36×106,0.57±0.22×106,and 0.34±0.18×106）.The proportion of CD4＋T cells and the cell count were markedly higher in the BCG-HAg group and the MS-HAg group than in the H37Ra-HAg group and the IL-2group（73.56±7.12% and66.93±7.65% vs.23.18±6.29% and 52.61±5.37%;4.75±0.86×106 and 3.28±0.59×106 vs.2.71±0.65×106and 1.63±0.45×106）.The proportion ofγδT cells and the cell count did not difference among the BCG-HAg group,the MS-HAg group,and the IL-2group. Conclusion H37Ra-HAg stimulates humanγδT cells,BCG-HAg and MS-HAg stimulate human CD4＋T cells,and BCG-HAg and MS-HAg do not stimulate humanγδT cells.

关 键 词：结核分枝杆菌 卡介苗 耻垢分枝杆菌 ΓΔT细胞 增殖 流式细胞术 

分 类 号：R378.911[医药卫生—病原生物学]