卡介苗与ESAT-6激活的γδT细胞表达抗原提呈细胞表型和功能的研究

发布时间：2019-02-26 19:31

【摘要】：目的:探讨卡介苗(BCG)与ESAT-6激活的人外周血γδT细胞是否具有抗原提呈细胞的表型和功能。方法:用Ficoll密度梯度离心法分离人外周血单个核细胞(PBMC)后,经尼龙毛柱法分离获得T细胞。将T细胞分别用卡介苗与ESAT-6刺激扩增γδT细胞后用流式细胞仪分选纯化γδT细胞,并检测其抗原提呈细胞表型。PBMC用贴壁法制备单核细胞后,诱导培养分化为成熟DC细胞,用流式细胞仪检测其成熟程度。T细胞用CFSE标记后分别按四种方法培养:①与结核分枝杆菌分泌性抗原(Mtb-Sag)共同培养;②与Mtb-Sag孵育2小时后的卡介苗激活的γδT细胞共同培养;③与Mtb-Sag孵育2小时后的ESAT-6激活的γδT细胞共同培养;④与Mtb-Sag孵育2小时后的成熟DC细胞共同培养;培养后检测T细胞及CD4+T细胞的增殖情况。结果:卡介苗与ESAT-6激活的γδT细胞CD80、CD86、HLA-DR的表达水平都增加,且后者显著高于前者。卡介苗与ESAT-6激活的γδT细胞培养组的T细胞增殖总数及CD4+T细胞的增殖有所增加;而后者显著高于前者,与成熟DC细胞培养组的增殖情况相似。结论:用卡介苗与ESAT-6激活的人外周血γδT细胞都具有抗原提呈细胞的表型和抗原提呈作用;但后者显著高于前者。
[Abstract]:Aim: to investigate the phenotype and function of antigen presenting cells in human peripheral blood 纬未 T cells activated by BCG (BCG) and ESAT-6. Methods: human peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density gradient centrifugation and T cells were isolated by nylon hair column method. 纬未 T cells were isolated and purified by flow cytometry after stimulated by BCG and ESAT-6 respectively, and their phenotypes of antigen presenting cells were detected. After monocytes were prepared by adherent method, the cells were induced to differentiate into mature DC cells, and the differentiation of 纬未 T cells into mature 纬未 T cells was observed by flow cytometry, and the differentiation of 纬未 T cells into mature PBMCs was observed. The mature degree of T cells was detected by flow cytometry. The T cells were cultured in four ways: 1) co-culture with Mycobacterium tuberculosis secretory antigen (Mtb-Sag); (2) 纬未 T cells activated by BCG were co-cultured with Mtb-Sag for 2 hours, 纬未 T cells activated by ESAT-6 were co-cultured with Mtb-Sag for 2 hours, and mature DC cells were incubated with Mtb-Sag for 2 hours. The proliferation of T cells and CD4 T cells was detected after culture. Results: both BCG and ESAT-6-activated 纬未 T cells increased the expression of CD80,CD86,HLA-DR, and the latter was significantly higher than that of the former. The total proliferation of T cells and the proliferation of CD4 T cells were increased in BCG and ESAT-6 activated 纬未 T cell culture groups, while the proliferation of CD4 T cells in the latter group was significantly higher than that in the mature DC cell culture group, which was similar to that of mature BCG T cell culture group. Conclusion: both BCG-activated 纬未 T cells and ESAT-6-activated human peripheral blood 纬未 T cells have the phenotypic and antigen presenting effects of antigen presenting cells, but the latter is significantly higher than the former.
【作者单位】：重庆医科大学附属第二医院呼吸内科;
【基金】：国家自然科学基金资助项目(No.30872261)
【分类号】：R392

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