Jin Li Ping, Li Da Jin, Wang Ming Yan, Zhu Xiao Yong, Zhu Ying, Ming Yi, Yuan Min Min
Laboratory for Reproductive Immunology, Institute of Obstetrics and Gynecology, Fudan University, Shanghai 200011, China.
Shi Yan Sheng Wu Xue Bao. 2005 Jun;38(3):199-204.
To study the effect of adoptive transfer of paternal antigen-tolerant T cells on recipient 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 proliferation of recipient splenocytes in response to DBA/2 stimulator cells and resulted in lower frequency of cells positive for IL-2, IFN-gamma, 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 transferred in recipient not only function as antigen-specific suppresser cells but also disable the recipient reactive T cells, which co-suppresses maternal rejection to the allogeneic fetus, thus resulting in the decrease of the embryo resorption rate of the abortion-prone mice to that of the normal pregnancy mice.
为研究父本抗原耐受T细胞的过继转移对受体反应性T细胞的影响,选用CBA/J×DBA/2交配作为易流产模型,CBA/J×BALB/c交配作为成功妊娠模型。妊娠第4天(着床时间),将易流产的CBA/J雌鼠与DBA/2雄鼠交配后,腹腔注射大鼠抗小鼠CD80和CD86单克隆抗体或大鼠同型IgG。妊娠第9天,用磁珠从妊娠CBA/J小鼠脾脏中获取纯化的T细胞,并在体外进行CFSE标记。妊娠第4天,将CFSE标记的T细胞静脉注射到其他与DBA/2雄鼠交配的CBA/J雌鼠体内。妊娠第9天,体外评估受体脾细胞对DBA/2刺激细胞的增殖反应,并用流式细胞术分析CFSE+/-T细胞中细胞内细胞因子和共刺激分子的表达。结果显示,与同型IgG处理的CBA/J小鼠的T细胞过继转移相比,父本抗原耐受T细胞或来自与BALB/c交配的CBA/J小鼠的T细胞的过继转移均显著抑制了受体脾细胞对DBA/2刺激细胞的增殖反应,并导致CFSE+CD3+群体和CFSE-CD3+群体中产生IL-2、IFN-γ、CD28的阳性细胞频率降低,产生IL-10、CTLA-4的细胞频率升高,而产生IL-4的细胞频率未出现显著变化。我们的研究结果表明,在受体中转移的父本抗原耐受T细胞不仅作为抗原特异性抑制细胞发挥作用,还使受体反应性T细胞失活,共同抑制母体对同种异体胎儿的排斥反应,从而导致易流产小鼠的胚胎吸收率降低至正常妊娠小鼠的水平。