5－氮－2′－脱氧胞苷诱导的调节性T细胞转输对自然流产孕鼠妊娠结局的影响

摘要

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.