雷帕霉素诱导Balb/c小鼠CD4~+ CD25~+ foxp3~+调节性T细胞增殖

目的探讨雷帕霉素对Balb/c小鼠CD4+ CD25+ foxp3+调节性T细胞的作用。方法取8wk龄的SPF级Balb/c小鼠30只,随机分为两组,实验组每只灌胃雷帕霉素0.4mg.d-1,对照组灌胃每天予等体积无菌水,共3wk。无菌条件下肝素抗凝心脏采血,分离脾脏,制备单细胞悬液。采用流式细胞仪检测小鼠外周血和脾细胞CD4+CD25+T细胞,实时定量PCR检测小鼠脾细胞foxp3 mRNA的表达。结果实验组小鼠外周血和脾细胞中CD4+CD25+T细胞的比例分别为(9.95±4.65)%和(24.13±10.06)%,对照组小鼠外周血和脾细胞中CD4+CD25+T细胞的比例分别为(5.01±1.49)%和(8.48±3.19)%,差异均有显著性(P<0.01)。实验组小鼠脾细胞foxp3 mRNA的表达水平明显高于对照组,是对照组的6.029倍,差异有显著性(P<0.01)。结论雷帕霉素能够明显诱导Balb/c小鼠体内CD4+CD25+T细胞的增殖,并能提高foxp3+ mRNA的表达。

Rapamycin induces Balb/c murine CD4+CD25+foxp3+T cells proliferations

Aim To investigate rapamycin(RAPA)induce Balb/c murine CD4+CD25+ foxp3+T cells proliferations.Methods Balb/c mice aging 8~10 weeks were used. RAPA was given to Balb/c murine 0.4 mg/day intragastric administration. The mice were given with sterile water as the control group, all mice were kept under specific pathogen-free conditions. drinking water and food were steriled. After three weeks, peripheral blood was collected and spleen cells were prepared, CD4+CD25+T cells were detected with FCM(CD4-pcy5, CD25-FITC), the relative levels of foxp3 mRNA were determined by Real-time quantitative RT-PCR in total splenocytes.Results The CD4+CD25+T cell of peripheral blood of experimental group and control group was (9.95±4.65)% and (5.01±1.49)%, respectively (P<0.01),and CD4+CD25+T cell of experimental mice splenocytes was (24.13±10.06)%,while control group was (8.48±3.19)% (P<0.01).Real-time quantitative RT-PCR showed that the levels of foxp3 mRNA of experimental mice splenocytes was 6.029 folds than that of control group(P<0.01).Conclusion RAPA can induce Balb/c murine CD4+CD25+T cells proliferation and enhanced foxp3+ mRNA in vivo,which suggested that RAPA could increase CD4+CD25+foxp3+T cells on the autoimmune disease and graft versus host disease treatment.