IL-17/IL-22在结核耐热性抗原和磷酸化抗原扩增极化后的表达及意义

Expression and significance of IL-17 and IL-22 in tuberculosis heat-resistant antigen and phospho-antigen stimulated cell amplified polarization

目的γδT细胞在结核耐热性抗原(Mtb-HAg)和磷酸化抗原(HDMAPP)培养扩增后,阐明Mtb-HAg和HDMAPP在细胞因子(ck:IL-1β、TGF-β和IL-23)极化状态下,IL-17和IL-22表达情况。方法密度梯度离心法获取外周血单个核细胞(PBMC),用RPMI1640培养基培养。实验组:分Mtb-HAg、HDMAPP、全菌抗原3组,各组分别加入(IL-1β、TGF-β)培养3 d加入rIL-2继续培养,第9天加入IL-23培养至12 d;对照组:分Mtb-HAg、HDMAPP、全菌抗原3组,培养3 d加入rIL-2继续培养至12 d。各组最终扩增为富含效应性γδT细胞的细胞群,收集细胞PMA/Ionomycin和Monensin刺激培养6 h,检测Tγδ17和Tγδ22细胞亚群进行分析。结果各组中γδT细胞均能产生IL-17和IL-22,Mtb-HAg+ck组较Mtb-HAg组及HDMAPP+ck组较HDMAPP组中γδT细胞产生IL-17细胞比例上升(均P<0.05),且IL-17主要由Vδ2亚群产生,而各极化实验组中γδT细胞产生IL-22细胞比例比较差异无统计学意义(均P>0.05)。结论 Mtb-HAg、HDMAPP刺激培养PBMC,在IL-1β、TGF-β和IL-23极化状态下,可诱导γδT细胞产生更多IL-17,而对诱导IL-22无效。

Objective To clarified the expression of IL-17 and IL-22 in tuberculosis heat-resistant antigen(Mtb-Hag)and phosphorylated antigen(HDMAPP)under the polarization state of IL-1β,TGF-βand IL-23 Happening.Methods Peripheral blood mononuclear cells(PBMC)were obtained by density gradient centrifugation and cultured in RPMI1640 medium.The experimental group was divided into three groups:Mtb-HAg,HDMAPP,and whole bacterial antigens.Each group was cultured with(IL-1β,TGF-β)for 3 days,rIL-2 for further culture,and IL-23 for 12 days on the 9 th day.The control group was divided into three groups:Mtb-HAg,HDMAPP,and whole cell antigen.After 3 days of culture,rIL-2 was added to continue to culture until 12 days.The experimental group and the control group finally expanded into a cell population rich in effectorγδT cells,then the cells were collected and stimulated with PMA/Ionomycin and Monensin for 6 hours,and Tγδ17 and Tγδ22 were detected and analyzed.The proportion of cell subpopulations was compared and analyzed.Results In all groups,γδT cells could produce IL-17 and IL-22.The percentage of IL-17 cells produced byγδT cells in the Mtb-Hag+ck group was significantly higher than that in the Mtb-HAg group,and the HDMAPP+ck group was higher than the HDMAPP group(all P<0.05).IL-17 was mainly produced by Vδ2 subpopulations,but there was no significant difference in the proportion of IL-22 cells produced byγδT cells in different polarization groups.Conclusion Under the polarization of IL-1β,TGF-βand IL-23,the cultured PBMC stimulated by Mtb-Hag and HDMAPP can induce more IL-17 production byγδT cells,but not IL-22.