摘要
目的体外观察雷帕霉素(Rapa)对转化生长因子-β1(TGF-β1)诱导的CD4+CD25+Treg细胞(i Treg)扩增的有效性及其相关性。方法以抗原提呈细胞(APC)、植物血凝素(PHA,10μg.L-1)、TGF-β1(7.5 ng.L-1)联合刺激诱导CD4+CD25-T细胞转化为CD4+CD25+Treg细胞,培养第5日添加不同浓度Rapa刺激培养3d作为实验组,设阴性对照组(不加Rapa)和空白对照组(RPMI 1640培养基),FCM检测各组中CD4+CD25+T细胞比例变化情况;MTT法测定各组诱导细胞的抑制功能。结果①阴性对照组CD4+CD25+T细胞比例较空白对照组增高(P<0.01),而实验组CD4+CD25+T细胞比例显著升高,Rapa(100 ng.mL-1)增殖效应显著(P<0.01),CD4+CD25+T细胞增殖比例达到42.6%;②实验组i Treg细胞能够明显抑制同型CD4+CD25-T细胞的增殖(P<0.01)且不同浓度Rapa之间存在差异,100 ng.mL-1Rapa抑制效应最明显(P<0.01)。结论在体外条件下,Rapa可以促进TGF-β1诱导分化的Treg细胞增殖,且具有免疫抑制功能,这为体外回输Treg细胞治疗疾病开辟了新途径。
Objective To investigate the effect of rapamycin on TGF-β1-induced Treg cells(iTreg) in vitro.Methods After CD4^+CD25^-T cells were stimulated with antigen-presenting cells(APC),PHA and TGF-β1 for 5 d,different levels of rapamycin were added into the cluture system(to establish an experimental group and a control group without rapamycin,while CD4^+CD25^-T cells co-cultured with RPMI 1640 were taken as a placebo group.After 3 days the stimulation percentage of CD4^+CD25^+T cells was detected by FCM and the inhibitory effect of proliferated iTreg was tested.Results ① The percentage of CD4^+CD25^+T cells in the negative control group was higher than that in the placebo control group(P〈0.01).The percentage of CD4^+CD25^+T cells in the experimental group markedly exceeded that of the negative control group(P〈0.01).After a 3-day culture,100 ng·mL-1 rapamycin was the optimum level that led to the most obvious expansion of iTreg cells(42.6%) in vitro.② Proliferation of CD4^+CD25^-T cells was suppressed by iTreg cultured with different levels of rapamycin(P〈0.01),and 100 ng·mL^-1 rapamycin had the most significant inhibitory effect.Conclusion Rapamycin can expand TGF-β1 induced Treg cells and iTregs can inhibit the proliferation of CD4^+CD25^-T cells,which is expected to open up a new way for Treg cells refusion in vitro.
出处
《中南药学》
CAS
2010年第6期437-441,共5页
Central South Pharmacy