过继转输的父系抗原耐受T细胞诱导受体母-胎免疫耐受的机制研究  

THE MECHANISM OF THE RECIPIENT MATERNAL-FETAL IMMUNO-TOLERANCE INDUCED BY THE TRANSFERRED PATERNAL ANTIGEN-TOLERANT T CELLS

作　　者：金莉萍[1] 李大金[1] 王明雁[1] 朱晓勇[1] 朱影[1] 孟毅[1] 袁敏敏[1] 

机构地区：[1]复旦大学附属妇产科研究所生殖免疫研究室,上海200011

出　　处：《实验生物学报》2005年第3期199-204,共6页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(39770773);复旦大学"985程"项目(985B36)资助

摘　　要：本文目的是为了探讨过继转输的父系抗原耐受T细胞对受体孕鼠反应性T细胞的影响。以(?)CBA/J×(?)BALB/c为正常妊娠模型,(?)CBA/J×(?)DBA/2为自然流产模型,将自然流产模型CBA/ J孕鼠于孕第4天(着床期)分别腹腔注射大鼠抗小鼠CD80和CD86 mAb或大鼠同型IgG。于孕第9天,应用免疫磁珠阴性分选孕鼠的脾脏T细胞,将纯化的T细胞先进行CFSE体外荧光标记,再分别过继转输至孕第4天的CBA/J×DBA/2孕鼠。于受体孕鼠孕第9天,在父系抗原刺激下,采用单向混合淋巴细胞反应分析受体孕鼠脾脏免疫细胞对父系抗原的增殖能力,并用流式细胞术分析过继转输的T细胞及受体T细胞内细胞因子IL-2、IL-4、IL-10和IFN-γ及细胞表面协同刺激分子CTLA-4的表达。本文结果显示,与过继转输父系抗原非耐受性T细胞相比,转输父系抗原耐受性T细胞和转输正常妊娠模型孕鼠T细胞均使受体孕鼠脾脏免疫细胞对父系抗原的增殖能力显著下降(P<0.05);同时使受体孕鼠T细胞内细胞因子IL-10及细胞表面CTLA-4的表达显著增加,而细胞因子IL-2和IFN-γ的表达则明显下降(P<0.05),细胞因子IL-4的表达不变(P>0.05)。总之,过继转输的父系抗原耐受T细胞具有抑制性免疫调节功能,诱导受体孕鼠反应性T细胞对父系抗原亦产生免疫耐受,两者协同作用则抑制母体对胚胎的免疫排斥,明显改善了自然流产模型孕鼠的妊娠预后。To study the effect of adoptive transfer of paternal antigen-tolerant T cells on re- cipient reactive T cells, CBA/JxDBA/2 mating was recruited as an abortion-prone model, and CBA/JxBALB/c mating as a successful pregnancy model. The abortion-prone CBA/J females mated with DBA/2 males were injected intraperitoneally with rat anti-mouse CD80 and CD86 mAb or rat isotype IgG at day 4 after gestation (time of implantation). The purified T cells were obtained from spleen of the pregnant CBA/J mice using magnetic beads at day 9 after gestation and labeled with CFSE in vitro. The CFSE-labeled T cells were intravenously injected into other CBA/J females mated with DBA/2 males at day 4 after gestation. The proliferation of recipient splenocytes in response to DBA/2 stimulator cells was evaluated at day 9 after gestation in vitro, and the expressions of intracellular cytokines and costimulatory molecules in CFSE^(+/-)T cells were analyzed by flow cytometry. The results showed that adoptive transfer of either paternal antigen- tolerant T cells or T cells from BALB/c-mated CBA/J mice significantly suppressed the prolifera- tion of recipient splenocytes in response to DBA/2 stimulator cells and resulted in lower frequen- cy of cells positive for IL-2, IFN-γ, CD28 and higher frequency of IL-10, CTLA-4-producing cells in both CFSE^+ CD3^+ population and CFSE^- CD3^+ population compared with adoptive transfer of T cells from isotype IgG-treated CBA/J mice, whereas the frequency of IL-4-producing cells did not appear significant change. Our findings suggest that paternal antigen-tolerant T cells trans- ferred in recipient not only function as antigen-specific suppresser cells but also disable the re- cipient reactive T cells, which co-suppresses maternal rejection to the allogeneic fetus, thus result- ing in the decrease of the embryo resorption rate of the abortion-prone mice to that of the nor- mal pregnancy mice.

关 键 词：T细胞 免疫耐受 抗原 父系 受体 机制研究 诱导 CTLA-4 IFN-γ IL-10 细胞因子 BALB/c 自然流产 免疫调节功能 增殖能力 免疫细胞 IL-2 IL-4 细胞表面 流式细胞术 CBA CD80 腹腔注射 免疫磁珠 荧光标记 反应分析 淋巴细胞 

分 类 号：R392[医药卫生—免疫学]