MtbAg体外激活扩增的人γδT细胞表面L-选择素的动态变化

The dynamics of L-selectin expression on activated human γδT cells by Mycobacterium tuberculosis antigen during in vitro proliferation

目的:研究结核杆菌抗原(MtbAg)激活扩增的人外周血γδT细胞表面L 选择素(CD6 2L)的表达。方法:用MtbAg和rIL-2刺激人外周血单个核细胞(PBMC)促使γδT细胞增殖,用CD3单克隆抗体(CD3mAb)和rIL-2刺激人PBMC促使αβT细胞增殖。取新鲜PBMC和刺激后培养3～2 1天的γδT细胞和αβT细胞,用多色荧光抗体标记,在流式细胞仪上检测γδT细胞表面CD6.2L的表达,并与αβT细胞作比较。结果:新鲜未刺激PBMC中γδT细胞CD6 2L表达率为4 3.9% ,明显低于αβT细胞的76 .6 % ;用MtbAg或CD3Ab激活后培养第6天时CD6 2L在γδT细胞的表达(90 .7% )和在αβT细胞表达(87.2 % )均达到高峰;以后CD6 2L表达逐渐下降,15～2 1天γδT细胞CD6 2L的表达在低水平(16 .2 %～10 .6 % )维持,αβT细胞表面的CD6 2L表达(2 3.6 %～17.5 % )亦在低水平维持。结论:新鲜PBMC中γδT细胞CD6 2L表达明显低于αβT细胞,但在激活后增殖过程中CD6 2L在γδT细胞的表达,与αβT细胞相似,呈现先上升再下降的趋势。

Objective:To study dynamic expression of L-selctin(CD62L) in human peripheral γδT cells activated with Mycobacterium tuberculosis antigen(MtbAg) and cultured for proliferation.Methods:Peripheral blood mononuclear cells(PBMC) were stimulated with MtbAg to generate MtgAg activated T cells(MtbAT),or with CD3mAb activated αβT cells(CD3AT).Fresh isolation PBMC and cultured MtbAT or CD3AT for 3 to 21 days were stained with multicolor fluorescence mAbs.The percentage of CD62L^+ cells among fresh PBMC,MtbAT or CD3AT were measured by flow cytometry.Results:The percentage of CD62L^+ cells of γδT cells(43.9%) in fresh PBMC has significant lower than that of αβT cells(76.6%),the expression of CD62L arrived at climax on day 6 of culture in γδT cells(90.7%) and αβT cells(87.2%),after then the expression of CD72L decreased and maintained low level,the percentages of CD62L^+ cells of γδU cells and αβT cells were 16.2% to 10.6% aod 23.6% to 17.5%,respectively,from day 15 to day 21 of culture.Conclusions:The CD62L expression of γδT cells in fresh PBMC was lower than that of αβTcells,but after activated by stimulation of MtbAg or CD3Ab,the expression of CD62L in human peripheral γδT cells,like αβTcells,rose in first and then decreased,and maintained low level later.