过继转输TSA诱导的CD4^＋CD25^＋调节性T细胞对不明原因复发性流产的治疗作用

目的观察过继转输TSA诱导的CD4^＋CD25^＋调节性T细胞（CD4＋CD25＋Treg）对不明原因流产的作用机制及妊娠预后的影响。方法以雌性CBA/J×雄性BALB/c为正常妊娠模型,以雌性CBA/J×雄性DBA/2J为自然流产模型,使用免疫磁珠方法分选雌性CBA/J小鼠脾脏CD4^＋CD25^＋Treg细胞,并使用流式细胞术检测分选纯度。采用TSA对流产孕鼠外周CD4^＋/CD25^-T细胞Foxp3基因特定位点进行表观修饰,以实现Foxp3稳定、持久的表达,并将CD4^＋Treg分别转输至流产模型孕4d（着床期）的雌性CBA/J孕鼠,于孕14d分别观察宿主孕鼠的胚胎吸收率。结果与对照组比较,过继转输TSA诱导的CD4^＋CD25^＋Treg细胞的宿主孕鼠的胚胎吸收率（11.27%）显著下降。结论孕早期过继转输TSA诱导的CD4^＋CD25^＋Treg细胞疗法能诱导宿主母胎免疫耐受,有利于妊娠的维持。

The treating function of TSA induced CD4~+CD25~+ regulatory T cells in unexplained recurrent spontaneous abortion

Objective:To investigate the mechanism and the effect of the adoptivetransfer of CD4+CD25+ regulatory T cells(CD4+CD25+ Treg)from the TSA induced mice on pregnant outcome of the abortion-prone mice.Methods: The experiments were performed in following two groups:using female CBA/J×male BALB/c matings as the normal pregnancy model and female CBA/J×male DBA/2J matings as the abortion-prone model.CD4+CD25+ Treg were isolated by MACS(magnetic cell sorting),FCM(flow cytometry)is to analyse the purity of CD4+CD25+ Treg.In order to achieve the stable and lasting expression of Foxp3,TSA is applied to the apparent heredity beautification of the special location of Gene Foxp3 in CD4+/CD25-T for miscarried mice.Purified CD4+CD25+ Treg cells were obtained from spleens of the virgin CBA/J mice and pregnant CBA/J mice at day 14 of gestation respectively,and then injected intravenously into the pregnant CBA/J mice mated with DBA/2J males at day 4 of gestation(time of implantation),then the embryo resorption rate was counted at day 14 of gestation.Results:Compare with control group,the embryo resorption rate(11.27%)after adoptive transfer of TSA induced pregnancy in CD4+CD25+Treg decreased significantly.Conclusion:The adoptive transfer of TSA-induced CD4+CD25+ regulatory T cells therapy can induce the maternal-fetal immuno-tolerance and maintaining pregnancy.