论文部分内容阅读
为了观察体外扩增的CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg)对不明原因复发性流产(RSA)妊娠预后的影响并探讨其作用机制。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,磁珠分选雌性CBA/J小鼠脾脏CD4~+CD25~+Treg并流式细胞术(flow cytometry,FCM)分析细胞纯度。妊娠第14天的CBA/J雌鼠(与雄性DBA/2J交配)为流产组;妊娠第14天的CBA/J雌鼠(与雄性BALB/c交配)为正常妊娠组;处女型CBA/J雌鼠脾CD4~+CD25~+Treg过继转输到孕4 d(着床期)CBA/J雌鼠(与雄性DBA/2J交配)体内为对照组;经体外培养扩增4 d的脾CD4~+CD25~+Treg过继转输到孕4 d(着床期)CBA/J雌鼠(与雄性BALB/c交配)体内为治疗组。分别于孕14 d分别观察宿主孕鼠的胚胎吸收率并测定宿主血清细胞因子IFN-γ/IL-10/TGF-β1表达水平和脾细胞叉状头转录因子Foxp3 mRNA相对含量。结果与对照组比较,过继转输体外扩增的CD4~+CD25~+Treg的宿主孕鼠的胚胎吸收率(11.32%)显著下降;治疗组中血清IL-10/IFN-γ、TGF-β1/IFN-γ比例有显著升高,外周血细胞因子IFN-γ有显著升高,细胞因子IL-10、TGF-β1无显著差异;Foxp3 mRNA相对表达量显著升高。孕早期经体外扩增的CD4~+CD25~+Treg疗法能诱导妊娠免疫耐受,有利于向妊娠维持的Th2、Th3、Tr1细胞偏移,叉状头转录因子Foxp3在妊娠免疫耐受上起着重要作用。
In order to observe the effect of expanded CD4 ~ + CD25 ~ + regulatory T cells (CD4 ~ + CD25 ~ + Treg) on the prognosis of unexplained recurrent spontaneous abortion (RSA) pregnancy and to explore its mechanism. Female CBA / J × male BALB / c as normal pregnancy model, female CBA / J × male DBA / 2J as spontaneous abortion model, magnetic beads were used to separate the splenic CD4 ~ + CD25 ~ + Treg from female CBA / J mice Cell purity was analyzed by flow cytometry (FCM). CBA / J females (mated with male DBA / 2J) on day 14 of gestation were miscarried; CBA / J females (mated with male BALB / c) on day 14 of gestation were in normal pregnancy; virgin CBA / J Female mice spleen CD4 ~ + CD25 ~ + Treg adoptive transfer to pregnant 4 d (implantation) CBA / J female mice (male DBA / 2J mating) as a control group in vivo; ~ + CD25 ~ + Tregs were adoptively transferred to pregnant rats (implantation stage). CBA / J female mice (mating with male BALB / c) were treated in vivo. The embryo resorption rate of the pregnant rats were observed respectively on the 14th day of pregnancy and the relative levels of the host serum cytokines IFN-γ / IL-10 / TGF-β1 and the splenic forkhead transcription factor Foxp3 mRNA were determined. Results Compared with the control group, the embryo absorptive rate (11.32%) of the pregnant pregnant mice with adoptively transferred CD4 + CD25 + Tregs was significantly decreased. The levels of IL-10 / IFN-γ, TGF- / IFN-γ ratio was significantly increased, peripheral blood cytokine IFN-γ was significantly increased, cytokines IL-10, TGF-β1 no significant difference; Foxp3 mRNA relative expression was significantly increased. In early pregnancy, CD4 ~ + CD25 ~ + Treg therapy induced by extracorporeal expansion can induce pregnancy immune tolerance, which is conducive to the migration of Th2, Th3 and Tr1 cells maintained by pregnancy. Forkhead-like transcription factor Foxp3 plays an important role in pregnancy immune tolerance An important role.