摘要
目的:建立MSCV-Foxp3感染法和TGF-β诱导法获取诱导性调节性T淋巴细胞(iTreg)的方法,并初步探讨两种iTreg的作用机制异同。方法:分别用MSCV-Foxp3感染和TGF-β诱导获取iTreg,流式细胞术和PCR检测两种iTreg的产率和Foxp3mRNA表达,CFSE染色观察iTreg的免疫抑制作用。ELISA检测iTreg的IL-10和TGF-β分泌,PCR检测iTreg颗粒酶A和B的mRNA水平。结果:感染法和诱导法iTreg产率分别可达49.12%和41.08%,但前者Foxp3 mRNA表达高于后者(P<0.05)。两种iTreg均具有一定的免疫抑制活性,IL-10分泌水平较原始CD4+T细胞上升(P<0.05),但颗粒酶B在诱导组iTreg的表达显著高于感染组(P<0.05)。结论:病毒感染法与TGF-β诱导法均可成功获得具有免疫抑制功能的iTreg,但前者Foxp3 mRNA表达高于后者。两种iTreg发挥免疫抑制功能可能都有IL-10参与,而颗粒酶B可能只与诱导组iTreg有关。
Objective： To establish the MSCV-Foxp3 infection method and TGF-β induced method to obtain induced regulatory T cells （iTreg）, and to investigate the similarities and differences between the two iTreg mechanisms. Methods： The iTreg eells were obtained by MSCV-Foxp3 and TGF-β respectively. Flow cytometry and PCR were used to detect iTreg yield and Foxp3 mRNA expression and CFSE staining was used to observe iTreg cell immunosuppression. ELISA was used to detect iTreg IL-10 and TGF-13 secretion levels. The iTreg granzyme A and B mRNA levels were detected by PCR. Results： The iTreg yield by infection method was up to 49.12%, and induced method up to 41.08%. The Foxp3 mRNA of the infected iTreg were markedly higher than those of the induced iTreg （P〈0.05）. Both kinds of iTreg couht significantly inhibit CD4＋ T cell proliferation and IL-10 secretion compared with non-Treg cells （P〈0.05）. Granzyme B expres- sion of the induced iTreg was significantly higher than that of the infected iTreg （P〈0.05）. Conclusion： MSCV-Foxp3 infection and TGF-β induction can be both used to obtain iTreg successfully, with the iTreg Foxp3 mRNA expression of the former being higher than that of the latter. 1 10 may be involved in both iTreg immunosuppression functions while granzvme B may only be involved in the TGF-β induced iTreg.
出处
《天津医科大学学报》
2013年第5期368-372,共5页
Journal of Tianjin Medical University
基金
高校博士点基金资助项目(20101202110001)
天津市应用基础与前沿计划重点项目(09JCZDJC19700)
973国家重点基础研究发展计划(2009CB918903)
广东省教育部产学研结合项目(2009B090300430)
