过继妊娠诱导的CD4^＋CD25^＋调节性T细胞治疗不明原因复发性流产  

Adoptive Pregnancy-induced CD4^＋CD25^＋ Regulatory T Cells Therapy in Unexplained Recurrent Spontaneous Abortion

作　　者：印永祥[1] 韩晓枫[2] 赵益明 何扬 

机构地区：[1]南京医科大学附属无锡市妇幼保健院病理科,江苏无锡214002 [2]南京医科大学基础医学院,江苏南京210029 [3]江苏省血液研究所,江苏苏州215006

出　　处：《中国血液流变学杂志》2009年第3期355-358,389,共5页Chinese Journal of Hemorheology

基　　金：2008年度无锡市卫生系统指令性科研项目（XMO802）

摘　　要：目的观察过继转输正常妊娠诱导的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 X雄性BALB／c为正常妊娠模型，以雌性CBA／J X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细胞并流式细胞术（flow cytometry，FCM）分析细胞纯度。将未孕CD4^＋CD25^＋Treg细胞和正常妊娠孕14d的CD4^＋CD25^＋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并测定宿主外周血和母胎界面组织体外培养上清液中Th1Th2／Th3型细胞因子（IFN-γ／TGF-β1／IL-10）水平。结果与对照组比较，过继转输正常妊娠诱导的CD4^＋CD25^＋Treg细胞的宿主孕鼠的胚胎吸收率（11．32％）显著下降；外周血TGF-β1／IFN-γ中位数（438）和IL-10／IFN-γ中位数（14）显著增加；母胎界面上TGF-β1中位数（5899．02pg／mL）和IL-10中位数（158．85pg／mL）艘著增加，IFN-γ中位数（18．46pg／mL）显著下降。结论孕早期过继转输正常妊娠诱导的CD4^＋CD25^＋Treg细胞疗法能诱导宿丰母胎免疫耐受并调节细胞因子表达，有利于妊娠维持的Th2、Th3应答偏移。Objective To investigate the mechanism and the effect of the adoptive transfer of CD4＋CD25＋ regulatory T cells（CD4＋CD25＋Treg） from the normal pregnancy mice on pregnant outcome of the abortion-prone mice.Methods The experiments were performed in following two groups：using female CBA/J x male BALB/c matings as the normal pregnancy model and female CBA/J x male DBA/2J matings as the abortion-prone model. Mouse CD4＋CD25＋Treg were isolated by MACS （magnetic cell sorting）,FCM （flow cytometry） is to analyse the purity of CD4^＋CD25^＋Treg.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）.The expression of Th 1,Th2 and Th3 cytokines （IFN-γ ,TGF-β 1,1L-10 respectively） at maternal-fetal interface and peripheral blood of the recipient mice was analysed,then the embryo resorption rate was counted at day 14 of gestation.Results Compared with control group,the embryo resorption rate（11.32%） after adoptive transfer of normal pregnancy-induced CD4^＋CD25^＋Treg decreased significantly.Median number of TGF-β 1/IFN-γ （438）and IL- 10/IFN-γ （14）at peripheral blood increased significantly.Median number of IL- 10（158.85 pg/mL） and TGF- 13 1 （5899.02pg/mL） at maternal-fetal interface increased significantly.Median number of IFN-γ （18.46 pg/mL） at maternal-fetal interface decreased significantly after adoptive transfer of pregnancy-induced CD4^＋CD25^＋ Treg from CBA/J x BALB/c matings. Conclusion The adoptive transfer of pregnancy-induced CD4^＋CD25^＋ regulatory T cells therapy can induce the maternal-fetal immuno-tolerance and regulate the expression ofcytokines to offset Th2,Th3 of maintaining pregnancy.

关 键 词：小鼠妊娠模型 CD4^+CD25^+调节性T细胞 过继转输 胚胎吸收率 细胞因子 

分 类 号：R392.9[医药卫生—免疫学]