Institute of Immunology of Shanxi Datong University, Datong, Shanxi, 037009, PR China.
Department of Hematology, The Fifth People's Hospital of Datong, Shanxi, 037009, PR China.
J Reprod Immunol. 2019 Feb;131:30-35. doi: 10.1016/j.jri.2018.12.002. Epub 2018 Dec 26.
To investigate whether epigenetic modification of CD4CD25 T-cells in vitro can make up for the inadequacy of CD4CD25Foxp3 Treg in animal model of spontaneous abortion and prevent immune response-mediated spontaneous abortion.
Trichostatin A (TSA) was applied to inhibit histone deacetylases (HDACs) and thereby to epigenetically modify the special location of Foxp3 gene in CD4CD25 T-cells of CBA/J mice. The expressions of CD25, Foxp3, CTLA-4 and PD-1 of CD4 T cells isolated from spleen of mice were characterized by flow cytometric analysis. Concentrations of transforming growth factor- β (TGF-β) and IL-10 in the supernatants of cultured Treg were measured using ELISA. The purified CD4 T cells treated with different reagents were injected into pregnant CBA/J mice mated with DBA/2J males on Day 1 and 4 of pregnancy, respectively. The embryo resorption rate was assessed on Day 14 of pregnancy.
TSA treatment significantly increased the population of CD4CD25Foxp3 iTreg. Those TSA induced Treg expressed high levels of PD-1 and CTLA-4, and secreted high levels of TGF-β and IL-10. Adoptive transfer of those iTreg at both early stage and implantation of stage of pregnancy significantly increased population of CD4CD25Foxp3 Treg in spleens of recipient miscarriage prone mice and significantly reduced resorption in those mice.
Epigenetic regulation of Foxp3 can generate functional regulatory T-cells. Adoptive transfer of TSA- induced CD4CD25Foxp3 Treg at an early stage of pregnancy can induce maternal-fetal immune tolerance and reduce embryo resorption in miscarriage prone mice.
探讨体外 CD4CD25T 细胞的表观遗传修饰是否能弥补自发性流产动物模型中 CD4CD25Foxp3Treg 的不足，从而防止免疫反应介导的自发性流产。
应用曲古抑菌素 A（TSA）抑制组蛋白去乙酰化酶（HDACs），从而对 CBA/J 小鼠 CD4CD25T 细胞中 Foxp3 基因的特定位置进行表观遗传修饰。采用流式细胞术分析分离自小鼠脾的 CD4T 细胞中 CD25、Foxp3、CTLA-4 和 PD-1 的表达。采用 ELISA 法检测培养的 Treg 上清液中转化生长因子-β（TGF-β）和白细胞介素-10（IL-10）的浓度。分别在妊娠第 1 天和第 4 天将用不同试剂处理的纯化 CD4T 细胞注射到与 DBA/2J 雄性交配的妊娠 CBA/J 小鼠体内。在妊娠第 14 天评估胚胎吸收率。
TSA 处理显著增加了 CD4CD25Foxp3iTreg 的群体。这些 TSA 诱导的 Treg 表达高水平的 PD-1 和 CTLA-4，并分泌高水平的 TGF-β和 IL-10。在妊娠早期和着床期将这些 iTreg 过继转移到易流产小鼠体内，显著增加了受体易流产小鼠脾脏中 CD4CD25Foxp3Treg 的群体，并显著降低了这些小鼠的吸收率。
Foxp3 的表观遗传调控可以产生功能性调节性 T 细胞。在妊娠早期转移 TSA 诱导的 CD4CD25Foxp3Treg 可以诱导母体-胎儿免疫耐受，减少易流产小鼠的胚胎吸收率。
