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目的探讨结核分枝杆菌耐热抗原(MTB-HAg)和抗T细胞受体γδ单克隆抗体(TCRγδm Ab)在刺激人γδT细胞诱导分化产生IL-17中的作用。方法健康人外周血单个核细胞经过免疫磁珠分选得到纯化的γδT细胞后,加入MTB-HAg或包被的TCRγδm Ab,再加或不加CD28 m Ab进行刺激,同时加或不加IL-17极化细胞因子组合(CK)(IL-1β、IL-6、TGF-β和IL-23),极化诱导培养10~12 d。然后收集诱导极化培养的γδT细胞,经佛波醇酯和离子霉素再刺激6 h后,用ELISPOT法检测产生IL-17的细胞数。结果经刺激和诱导分化培养的γδT细胞用ELISPOT法检测产生IL-17的细胞数发现,TCRγδm Ab联合CK组与TCRγδm Ab组相比明显增加,TCRγδm Ab联合CD28 m Ab组与TCRγδm Ab同时联合CD28 m Ab和CK组相比明显减少。TCRγδm Ab同时联合CD28 m Ab和CK组与TCRγδm Ab联合CK组相比明显增多。MTB-Hag同时联合CD28 m Ab和CK组与TCRγδm Ab同时联合CD28 m Ab和CK组相比显著减少,但与MTB-HAg联合CD28 m Ab组相比明显增多。结论诱导Th17分化的CK(IL-1β、IL-6、TGF-β和IL-23)也能诱导人γδT细胞分化产生IL-17,TCRγδm Ab刺激γδT细胞增殖后诱导分化产生IL-17的活性比MTB-HAg更强。另外,CD28在TCRγδm Ab激活γδT细胞诱导产生IL-17的过程中也有重要作用。
Objective To investigate the role of MTB-HAg and TCRγδm Ab in stimulating the differentiation of human γδT cells into IL-17. Methods The purified human γδT cells were obtained from the peripheral blood mononuclear cells of healthy people by immunomagnetic bead sorting and then stimulated with MTB-HAg or coated TCRγδm Ab with or without CD28 m Ab, with or without IL- Polarized cytokine combination (CK) (IL-1β, IL-6, TGF-β, and IL-23) was induced by polarization for 10-12 days. After that, γδT cells induced by polarized culture were collected and restimulated with phorbol ester and ionomycin for 6 hours. The number of IL-17 producing cells was detected by ELISPOT. Results The number of IL-17-producing cells was detected by ELISPOT. The results showed that TCRγδm Ab combined with CK increased significantly compared with TCRγδm Ab group. TCRγδm Ab combined with CD28 m Ab group and TCRγδm Ab combined with CD28 m Ab significantly reduced compared with CK group. TCRγδm Ab combined with CD28 m Ab and CK groups increased significantly compared with TCRγδm Ab combined with CK group. MTB-Hag combined with CD28 m Ab and CK groups and TCRγδm Ab combined with CD28 m Ab significantly decreased compared with CK group, but significantly increased compared with MTB-HAg and CD28 m Ab group. CONCLUSION: Th17-differentiated CK (IL-1β, IL-6, TGF-β and IL-23) can induce the differentiation of human γδT cells into IL-17 and TCRγδm Ab to stimulate the proliferation of γδT cells and induce IL-17 production Stronger than MTB-HAg. In addition, CD28 also plays an important role in the induction of IL-17 production by γδT cells by TCRγδm Ab.