IFN-γ和IL-17在结核病患者γδ T细胞中的表达及意义

Expression and significance of IFN-γ and IL-17 in γδ T cells of tuberculosis patients

目的探讨正常人(HC)、结核病患者(TB)2组人群外周血结核杆菌耐热性抗原(Mtb-HAg)刺激后γδ T细胞产生IFN-γ和IL-17的变化及意义。方法分别选取12例HC作为对照组,12例TB作为实验组,采集2组人群外周血进行分离PBMC,经Mtb-HAg刺激扩增培养后,收集Mtb-HAg体外扩增效应性T细胞,Mtb-HAg作为刺激剂,设定阴性对照组和实验组分别刺激培养2 h,再加入莫能霉素(monensin)阻断培养4 h,收集细胞加入荧光抗体进行表面分子和胞内染色,上流式细胞仪检测对照组和实验组γδ T细胞产生IFN-γ和IL-17细胞的相对比例。结果 HC组、TB组各阴性对照组和实验组γδ T细胞均可以产生IFN-γ,用Mtb-HAg再刺激时,γδ T细胞中产生IFN-γ的细胞均显著高于未用Mtb-HAg刺激组(P <0. 001),且HC组和TB组差异有统计学意义(P <0. 001);HC组γδ T细胞中产生IL-17的细胞呈现降低趋势,与阴性对照组比较差异无统计学意义,而TB组γδ T细胞中产生IL-17的细胞也呈现降低趋势,且差异有统计学意义(P <0. 01)。结论 TB患者PBMC培养的γδ T细胞对Mtb-HAg刺激有较强的反应能力,而产生IL-17的能力降低。

Objective To investigate the effects of Mtb-HAg on the production of IFN-γ and IL-17 in γδ T cells from peripheral blood of peripheral blood mononuelear cell （PBMCs） from normal people （HC） and tuberculosis patients （TB）. Methods PBMCs were isolated from the peripheral blood of 12 HC and 12 TB patients and cuhured for 12 days. The cells in the experimental group were stimulated by using Mtb-HAg for 2 h and then further cultured for 4 h in Monensin ; while the cells in the negative control group without receiving the stimulation with Mtb-HAg. The cells were harvested and labeled with fluorescent-labeled McAb for surface and intracellular staining. The γδ T cells in both groups were assayed by flow cytometry for the production of IFN-γ and IL-17 cells relative proportions. Results The γδ T cells in both experimental group and negative control group were able to produce IFN-γ. IFN-γ produced by γδ T cells stimulated by Mtb- HAg was significantly higher than that by cells without receiving stimulation with Mtb-HAg （P 〈 0.001 ）, and there was significant difference between TB group and HC group （P 〈 0.001 ）. In HC group,the rate of γδ T cells producing IL-17 decreased in both experimental group and negative control group without statistical significance difference, however, in TB group, the same trend showed significant difference between the experimental group and negative control group. Con- clusion The γδ T ceils from PBMCs in TB patients are sensitive to the stimulation with Mtb-Hag, but its ability to pro- duce IL-17 decreases obviously.