目的:研究过继转输5-氮-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5AzaD)诱导的 CD4+CD25+Foxp3+调节性T细胞(inducible regulatory T cells,iTregs)对自然流产孕鼠妊娠预后的影响。方法:以60例雌性CBA/J×雄性DBA/2J小鼠自然流产模型为研究对象,随机选取20例孕鼠经免疫磁珠分选脾脏CD4+T细胞,应用5AzaD对CD4+CD25-T细胞进行体外处理得到iTregs,通过流式细胞术检测表观修饰前后Tregs细胞中Foxp3的表达。分别向妊娠第1天和第4天的流产孕鼠体内输注iTregs作为治疗组,于妊娠14天比较各组孕鼠的胚胎吸收率。结果:经5AzaD修饰后,iTregs占CD4+T细胞的比例为(41.50±8.03)%。治疗组中妊娠第1天和第4天过继转输iTregs孕鼠的胎盘吸收率分别为10.47%和21.69%(P<0.05)。结论:对CD4+CD25-T细胞的表观修饰可能解决自然调节性T细胞来源不足的问题,在妊娠早期转输5AzaD诱导的iTreg细胞更有利于维持正常妊娠。
Objective:To investigate the effect of the adoptive transfer of CD4+CD25+Foxp3+regulatory T cells ( iTregs) induced by 5-aza-2′-deoxycytidine (5AzaD) on pregnant outcome of the abortion-prone mice.Methods:Sixty cases of female CBA/J male DBA/2J abortion-prone matings were taken as study group,the CD4+T cells from spleen of twenty female CBA/J mice were separated by magnetic activated cell sorting (MACS),5AzaD was applied to the conversion of CD4+CD25-T cells to iTregs,the expression of Foxp3 in Tregs was characterized by flow cytometry analysis before and after epigenetic modification.The purified iTregs were injected into abortion-prone mice on day 1 or 4 of pregnancy,respectively,which were used as therapy groups,and then the embryo resorption rate was counted on day 14 of pregnancy.Results:After the treatment of 5AzaD,the percentage of iTregs in CD4+T cells was (41.50±8.03)%.The embryonic absorption rates of the two therapy groups were 10.47%(on day 1 of pregnancy) and 21.69%(on day 4 of pregnancy) ,respectively ( P〈0.05 ) .Conclusion: Epigenetic modication of CD4+CD25-T cells may solve the problem of nTregs deficiency,particularly adoptive therapy of 5AzaD-induced iTregs at early stage of pregnancy can maintain normal pregnancy.