上海交通大学学报(医学版)

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过继性转移TGF-β诱导CD4+CD25+调节性T细胞对自然流产模型小鼠胚胎丢失率的影响

邱 添,滕银成,徐 亮   

  1. 上海交通大学附属第六人民医院妇产科, 上海 200233
  • 出版日期:2014-08-28 发布日期:2014-09-02
  • 通讯作者: 徐 亮, 电子信箱: xuliang19770323@126.com。
  • 作者简介:邱 添(1983—), 女, 住院医师, 硕士; 电子信箱: t_princessonly@hotmail.com。
  • 基金资助:

    国家自然科学基金青年项目(81100474)

Effects of CD4+CD25+ regulatory T cells induced by adoptive transfer of TGF-β on rate of embryo loss of spontaneous abortion mice model

QIU Tian, TENG Yin-cheng, XU Liang   

  1. Department of Obstetrics and Gynecology, the Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China
  • Online:2014-08-28 Published:2014-09-02
  • Supported by:

    National Natural Science Foundation of China, 81100474

摘要:

目的 观察过继性转移转化生长因子-β (TGF-β) 诱导生成CD4+CD25+调节性T细胞对自然流产模型小鼠胚胎丢失率的影响。方法 从正常妊娠CBA/J孕鼠脾脏中分选
CD4+CD25T细胞,体外经TGF-β诱导转化为CD4+CD25+调节性T细胞,流式细胞仪检测其对自身反应性T细胞增殖的抑制作用。实验小鼠随机分为正常妊娠组和自然流产模型组,后者根据干预方式的不同分为空白组(自然流产模型未干预)、CD4+CD25+T细胞组(自然流产模型过继性转移天然CD4+CD25+调节性T细胞)、TGF-β诱导CD4+CD25+T细胞组(自然流产模型过继性转移TGF-β诱导CD4+CD25+调节性T细胞)和对照组(自然流产模型注射CD4+CD25T细胞)。于孕第14日采集血样后处死小鼠。 ELISA法检测血清白介素-10 (IL-10)、TGF-β1和γ干扰素(IFN-γ)水平,Western blotting和Real-time PCR检测蜕膜组织Foxp3 mRNA和蛋白的表达。计算各组孕鼠胚胎丢失率。结果 与空白组和对照组比较,天然CD4+CD25+T细胞组和TGF-β诱导CD4+CD25+T细胞组血清IL-10、TGF-β1水平升高,IFN-γ水平降低;蜕膜组织中Foxp3 mRNA和蛋白的表达上调;胚胎丢失率显著降低(P<0.05)。结论 向自然流产模型小鼠过继性转移TGF-β诱导生成CD4+CD25+调节性T细胞,可诱导小鼠体内Th1/Th2免疫反应向Th2优势偏移,降低胚胎丢失率。

关键词: 自然流产, 过继性转移, 转化生长因子-β, CD4+CD25+调节性T细胞, Foxp3

Abstract:

Objective To observe the effects of CD4+CD25+ regulatory T (Treg) cells induced by adoptive transfer of transforming growth factor-β (TGF-β) on the rate of embryo loss of mice of the spontaneous abortion model. Methods CD4+CD25- cells were isolated from the spleen of normal pregnant CBA/J mice and induced to CD4+CD25+ Treg cells by TGF-β in vitro. The inhibit effect of TGF-β induced Treg cells on the proliferation of self-reactive T cells was measured by the flow cytometry. The CBA/J mice were randomly divided into the normal pregnancy group and spontaneous abortion group. The latter was further divided into the blank group (spontaneous abortion model without intervention), innate CD4+CD25+T cells group (spontaneous abortion model with adoptive transfer of innate CD4+CD25+ Treg cells), TGF-β induced CD4+CD25+ T cells group (spontaneous abortion model with adoptive transfer of CD4+CD25+ Treg cells induced by TGF-β), and control group (spontaneous abortion model with injection of CD4+CD25- T cells) according to the intervention method. On day 14 of pregnancy, the mice were sacrificed and the blood samples were gathered. The levels of serum IL-10, TGF-β1, and IFN-γ were measured by the ELISA. The expressions of Foxp3 mRNA and protein in deciduas tissues were measured by the Western blotting and Realtime PCR. The rate of embryo loss of each group was calculated. Results Compared to the blank group and control group, the levels of serum IL-10 and TGF-β1 of the innate CD4+CD25+T cells group and TGF-β induced CD4+CD25+T cells group increased; the level of serum IFN-γ decreased; the expressions of Foxp3 mRNA and protein in deciduas tissues increased; and the rate of embryo loss significantly decreased (P<0.05). Conclusion CD4+CD25+ Treg cells induced by adoptive transfer of TGF-β for mice of the spontaneous abortion model can induce Th1 and Th2 immune responses towards Th2 and decrease the rate of embryo loss.

Key words: spontaneous abortion, adoptive transfer, TGF-β, CD4+CD25+regulatory T cell, Foxp3