论文部分内容阅读
目的研究雷帕霉素(Rapa)对同种移植耐受个体CD4+CD25+T细胞体内负免疫调节作用的影响。方法建立同种皮肤移植模型,受体鼠术前预输注供体鼠脾细胞,术后给予环孢素A(CsA)进行耐受诱导。移植后第14天提取耐受诱导模型鼠的T细胞,经不同浓度Rapa和/或IL-2体外处理后,混合淋巴细胞反应(MLR)确定T细胞特异增殖水平;流式细胞术(FCM)检测CD4+CD25+T细胞比例变化;RT-PCR检测Foxp3mRNA表达情况;ELISA检测细胞培养不同时间后上清中IL-10的变化。然后将Rapa和/或IL-2处理的T细胞过继转移给同种移植后的BALB/c-SCID鼠,观察移植物存活状态。结果CsA加供体脾细胞预先注射可明显延长小鼠移植皮片的存活期(P<0.05);移植耐受状态的T细胞经Rapa和/或IL-2体外处理后CD4+CD25+T细胞比例升高、增殖水平明显降低、Foxp3表达量明显增加;过继转输给同种移植SCID鼠后,其移植皮片存活时间显著延长(P<0.05)。结论Rapa可体外扩增耐受诱导模型中CD4+CD25+T细胞,使CD4+CD25+T细胞相关的Foxp3和IL-10明显升高,过继免疫后,小鼠同种移植物存活时间明显延长,而低浓度IL-2可以协同Rapa的这一作用。
Objective To investigate the effect of rapamycin on the negative immune regulation of CD4 + CD25 + T cells in allograft tolerant individuals. Methods Allogeneic skin graft model was established. Precontracted donor spleen cells were pretreated with CsA in mice. On the 14th day after transplantation, T lymphocytes were isolated from T cells of tolerance-induced model mice and treated with different concentrations of Rapa and / or IL-2 in vitro. Mixed lymphocyte reaction (MLR) was used to determine T-cell specific proliferation. Flow cytometry (FCM) The proportion of CD4 + CD25 + T cells was detected. The expression of Foxp3 mRNA was detected by RT-PCR and the level of IL-10 in supernatants was detected by ELISA. Rapa and / or IL-2-treated T cells were then adoptively transferred to allograft BALB / c-SCID mice and graft survival was observed. Results Preadventive injection of CsA plus donor spleen cells significantly prolonged the survival of transplanted skin grafts in mice (P <0.05). The T cells tolerated by transplants were treated with Rapa and / or IL-2 in vitro and the CD4 + CD25 + T cells The proportion of Foxp3 expression was significantly increased after transplanted into SCID mice. The graft survival time was significantly prolonged (P <0.05). Conclusion Rapa can expand CD4 + CD25 + T cells in tolerance-induced model in vitro and significantly increase Foxp3 and IL-10 associated with CD4 + CD25 + T cells. After adoptive immunization, the survival time of mice allografts was significantly prolonged , While low concentrations of IL-2 synergize Rapa’s role.