雷帕霉素对诱导nave小鼠效应性T细胞转化为调节性T细胞的影响

Effect of Rapamycin in Inducing Nave Murine Effector T Cell Convert to Regulatory T Cell

目的研究雷帕霉素对诱导naive小鼠效应性T细胞（Teff）体外转化为调节性T细胞（Treg）的影响。方法取6-8周龄C57BL/6小鼠的脾及淋巴结,分离提纯叉头蛋白3（FoxP3）阴性的Teff,分为与成熟树突状细胞（mDC）、B细胞和抗CD3抗体（Anti-CD3）共培养3组,前两组6 d后收获细胞,Anti-CD3组4 d后收获细胞。每组设对照组与实验组,对照组未加入雷帕霉素,实验组加入雷帕霉素,使其浓度分别为1、10、50、100 nmol/L,流式细胞仪检测收获细胞中FoxP3阳性的Treg比率。结果FoxP3阳性的Treg细胞比率：在mDC组中对照组为0.01%,实验组分别为0.39%、0.47%、0.34%、0.26%;在B细胞组中对照组为0.01%,实验组分别为5.56%、5.89%、7.15%、4.72%;在Anti-CD3组中对照组为0.93%,实验组分别为1.35%、1.07%、1.02%、1.19%。mDC组及Anti-CD3组各实验组与对照组相比差异无显著性,B细胞组中各实验组与对照组相比差异具有显著性（P〈0.01）。结论在体外以B细胞作为抗原呈递细胞,雷帕霉素能够诱导naive小鼠Teff转化为FoxP3阳性的Treg。

Objective To study the effect of rapamycin in inducing naive murine effector T cell（Teff） convert to regulatory T cell（Treg） in vitro.Methods The forkhead box protein 3（FoxP3） negative Teff were isolated and purified from the spleen and lymph node of C57BL/6 murines aged 6-8 weeks,then Teff were cultured in three groups with mature dendritic cells（mDC）,B cells,and plate coated Anti-CD3.In addition,the control wells and the test wells were prepared in each group,rapamycin were not added in the control wells but added in the test wells with concentrations of 1,10,50,and 100 nmol/L.Percentages of FoxP3 positiveTreg were examined by flow cytometry after 4 days in Anti-CD3 group and after 6 days in the other two groups.Results As shown by the flow cytometry,the percentages of Fox P3 positive Treg were as follows in three group： in the mDC group,it was 0.01% in the control well and 0.39%,0.47%,0.34%,and 0.26% in test wells;in B cell group,it was 0.01% in the control wells and 5.56%,5.89%,7.15%,and 4.72% in the test wells;in Anti-CD3 group,it was 0.93% in the control wells and 1.35%,1.07%,1.02%,and 1.19% in test wells.No significant difference was found between the test wells and control wells in the mDC group and Anti-CD3 group;however,the percentages of FoxP3 positive Treg was significantly different between the test wells and control wells in the B cell group（P〈0.01）.Conclusion When B cell is acted as the antigen-presenting cell,rapamycin can effectively induce Teff convert to Treg in vitro.