Foxp3基因转染小鼠CD4+CD25-T细胞对同种T细胞增殖反应的影响

目的观察Foxp3基因转染的小鼠CD4 CD25-T细胞对同种T细胞增殖反应的影响。方法以免疫磁珠法分离小鼠脾细胞中的CD4 CD25 和CD4 CD25-T细胞,用流式细胞术检测细胞纯度,以电穿孔法将质粒PMFOXP3-IRES2-EGFP转染小鼠CD4 CD25-T细胞,用荧光显微镜观察转染细胞中Foxp3的表达,转染细胞和CD4 CD25 T细胞与同种T细胞做混合淋巴细胞培养,以3H-TdR掺入测定CPM值。结果在质粒PMFOXP3-IRES2-EGFP转染CD4 CD25-T细胞24h后,转染细胞中可检测到高水平的Foxp3;转染细胞和CD4 CD25 T细胞CPM值分别为3 927.0±674.7和3 895.3±504.6(P>0.05)。结论以质粒PMFOXP3-IRES2-EGFP转染的CD4 CD25-T细胞对同种T细胞增殖反应具有显著抑制作用,与CD4 CD25 T细胞的抑制作用等同。

Influence of Foxp3 gene-transferred CD4+CD25- T cells on proliferation of allo T cells

Objective To observe the immunosuppressive efficacy of mice CD4~ CD25~-T cells transfected with Foxp3 gene.Methods Mice CD4~ CD25~-and CD4~ CD25~ regulatory T cells were isolated by magnetic beads from the spleen.The expression of CD4 and CD25 on the cells was determined by means of flowcytometry.The isolated CD4~ CD25~-T cells were transferred with plasmid PMFOXP3-IRES2-EGFP by electroporation.The expression of Foxp3 was determined with immune fluoresce nt microscopy.The function of Foxp3 gene transferred T cells was tested by MLC.Results A high expression of Foxp3 was foune in Foxp3 gene-transferred T cells 24h after transferring.The MLC was suppresed significantly by adding of Foxp3 gene-transferred T cells.Conclusion The CD4~ CD25~-T cells transferred with plasmid PMFOXP3-IRES2-EGFP have the same immunosuppressive efficacy with CD4~ CD25~ regulatory T cells.