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

Effect of adoptive transfer of 5Aza D- induced regulator T cells on outcomes of gestation in abortion-prone murine matings

目的:研究过继转输5-氮-2'-脱氧胞苷(5-aza-2'-deoxycytidine,5Aza D)诱导的CD4+CD25+Foxp3+调节性T细胞(inducible regulatory T cells,i Tregs)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5Aza D对CD4+CD25-T细胞进行体外处理得到i Tregs,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i Tregs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5Aza D修饰后,i Tregs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i Tregs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5Aza D诱导的i Treg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.