Expression and significance of IL-17 and IL-22 in tuberculosis heat-resistant antigen and phospho-antigen stimulated cell amplified polarization
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摘要:
目的 γδ T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γδ T细胞的细胞群，收集细胞PMA/Ionomycin和Monensin刺激培养6 h，检测Tγδ 17和Tγδ 22细胞亚群进行分析。 结果 各组中γδ T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γδ T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γδ T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γδ T细胞产生更多IL-17，而对诱导IL-22无效。 Abstract: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-β and 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 9th 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 effector γδ T cells, then the cells were collected and stimulated with PMA/Ionomycin and Monensin for 6 hours, and Tγδ 17 and Tγδ 22 were detected and analyzed. The proportion of cell subpopulations was compared and analyzed. Results In all groups, γδ T cells could produce IL-17 and IL-22. The percentage of IL-17 cells produced by γδ T 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 by γ δ T cells in different polarization groups. Conclusion Under the polarization of IL-1 β, TGF-β and IL-23, the cultured PBMC stimulated by Mtb-Hag and HDMAPP can induce more IL-17 production by γδ T cells, but not IL-22. -
Key words:
- Tuberculosis /
- Mtb-HAg /
- HDMAPP /
- IL-17 /
- IL-22
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图 1 各组γδT细胞分泌IL-17和IL-22细胞的比例情况
注：Mtb-HAg+ck组与Mtb-HAg组比较，aP < 0.05；HDMAPP+ck组与HDMAPP组比较，bP < 0.05。
表 1 Mtb-HAg组与其极化组Vδ1和Vδ2亚群产生IL-17细胞的比较(x ±s, %)
组别 Vδ1亚群 Vδ2亚群 Mtb-HAg 0.11±0.07 1.10±0.82 Mtb-HAg+ck 0.16±0.05 2.97±1.52 t值 -1.479 -2.416 P值 0.170 0.042 
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表 2 HDMAPP组与其极化组Vδ1和Vδ2亚群产生IL-17细胞的比较(x ±s, %)
组别 Vδ1亚群 Vδ2亚群 HDMAPP 0.13±0.04 0.47±0.37 HDMAPP+ck 0.21±0.08 3.12±1.32 t值 -2.112 -5.769 P值 0.071 0.014
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