摘要
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。
To investigate the effect and the mechanism of amplification of the CD4＋ CD25＋ regulatory T cells （CD4＋ CD25＋ Tregs） in vitro on pregnant outcome of the abortion-prone mice experiments was performed in two animal models; one of which using female CBA/J male BALB/c matings as the normal pregnancy model and the other using female CBA/J male DBA/ 2J matings as the abortion-prone model. Mouse CD4＋ CD25＋ Tregs were isolated by magnetic cell sorting（MACS） and the pu- rity of CD4＋ CD25＋ Tregs was confirmed by flow cytometry（FCM） for analysis. We set up 4 groups in this experiment： （1） the pregnant CBA/J mice mated with DBA/2J males at day 14 of gestation was used as the abortion group; （2） the pregnant CBA/J mice mated with BALB/c males at day 14 of gestation as normal pregnancy group; （3） the purified CD4＋ CD25＋ Tregs were obtained from spleens of the virgin CBA/J mice and then injected intravenously into the pregnant CBA/J mice mated with DBA/2J males at day 4 of gestation（time of implantation）. This group of mice was used the as control group; （4） the amplification of CD4＋ CD25＋ Tregs at 4 days in vitro were obtained from spleens of the virgin CBA/J mice and then injected intrave- nously into the pregnant CBA/J mice mated with DBA/2J males at day 4 of gestation（time of implantation）. This group of mice was used as therapy group. The expressions of cytokines （IFN-γ, TGF-131, IL-10）in peripheral blood of the recipient mice were analyzed and the embryo resorption rate was counted at day 14 of gestation. Reverse transcription-polymerase chain reaction（RT-PCR）was used to examine the expression of Foxp3 mRNA in the spleen cells from these 4 groups of mice. Compared with the control group, the embryo resorption rate of mice under treatment was decreased significantly; while the expressions of TGF-β1/IFN-γ and IL-10/IFN-γ in peripheral blood increased significantly. Meanwhile, the expression of IFN-γ in peripheral blood was significantly decreased but
出处
《现代免疫学》
CAS
CSCD
北大核心
2010年第2期132-137,共6页
Current Immunology
基金
2008年度无锡市卫生系统指令性科研资助项目(XM0802)
