调节性T细胞治疗实验性小鼠多发性肌炎及机制研究

Regulatory T cells in the treatment of autoimmune myositis in mice: efficacy and mechanism

目的探讨CD4+CD25+Foxp3+Treg细胞对于实验性自身免疫性肌炎(EAM)小鼠的治疗价值及机制。方法免疫磁珠分选技术从BALC/c小鼠脾脏中分选出足量CD4+CD25+Foxp3+Treg细胞以备回输,观察干预组及未干预组EAM小鼠肌肉组织病理学变化,流式细胞仪检测两组小鼠脾脏CD4+CD25+Foxp3+Treg细胞表面PD-1及CTLA-4的表达变化,双抗体夹心ELISA法检测外周血IL-10、TGF-β的变化。结果干预组小鼠较未干预组肌肉病理炎性细胞浸润明显减轻,其外周血IL-10、TGF-β含量较未干预组明显升高(P<0.05),脾脏CD4+CD25+Foxp3+Treg细胞表面PD-1及CTLA-4表达明显升高(P<0.05)。结论 CD4+CD25+Foxp3+Treg细胞回输对于EAM小鼠的治疗效果是通过增加外周血IL-10及TGF-β水平和增高脾脏CD4+CD25+Foxp3+Treg细胞表面PD-1及CTLA-4的表达发挥作用。

Objective To investigate effect of CD4＋CD25＋Foxp3＋Tregs in the treatment of autoimmune myositis （EAM） in mice and explore the possible mechanisms. Methods Mouse models of EAM were divided randomly into model group and treatment group, and the latter received infusion of CD4 ＋ CD25 ＋ Foxp3 ＋ Tregs separated from normal mouse spleen by magnetic activated cell sorting. The changes of muscle pathology was observed, and the expression of PD-1 and CTLA-4 in spleen CD4＋ CD25＋ Foxp3＋ Tregs was analyzed using flow cytometry; peripheral blood IL-10 and TGF-β levels were tested using double antibody sandwich ELISA. Results Compare with the model group, the mice in the treatment group showed significantly reduced muscular inflammatory cell infiltration, increased blood levels of IL-10 and TGF-β （P〈0.05）, and increased expression of PD-1 and CTLA-4 in spleen CD4＋CD25＋Foxp3＋Tregs （P〈0.05）. Conclusion CD4＋CD25＋Foxp3＋Tregs reinfusion produces therapeutic effect in mice with EAM by increasing peripheral blood IL-10 and TGF-βlevels and PD-1 and CTLA-4 expressions in spleen CD4＋CD25＋Foxp3＋Tregs.