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目的探讨Foxp3基因转染对大鼠CD4+CD25-T淋巴细胞体外增殖能力的影响。方法以逆转录病毒载体介导Foxp3基因进入大鼠CD4+CD25-T淋巴细胞,以TGF-β诱导Foxp3基因表达。通过观察报告基因的表达水平及流式细胞仪检测评价转染效率,将转染Foxp3基因的大鼠CD4+CD25-T淋巴细胞、CD4+CD25+T淋巴细胞、大鼠脾脏单个核细胞分别与同系大鼠单个核细胞混合培养,液体闪烁法测定各组细胞增殖水平。结果Foxp3组增殖水平(3 354.18±203.73)略高于调节性T细胞组(2 919.25±137.91),但明显低于空白对照组(12 706.06±967.47)。结论经逆转录病毒介导Foxp3基因可在CD4+CD25-T淋巴细胞高表达,并使该细胞获得与CD4+CD25+T淋巴细胞相似的免疫无能及免疫抑制特性。
Objective To investigate the effect of Foxp3 gene transfection on the proliferation of CD4 + CD25-T lymphocytes in vitro. Methods The Foxp3 gene was transduced into rat CD4 + CD25-T lymphocytes by retroviral vector to induce Foxp3 gene expression by TGF-β. By observing the expression level of reporter gene and evaluating the transfection efficiency by flow cytometry, the expression of Foxp3 gene in rat CD4 + CD25-T lymphocytes, CD4 + CD25 + T lymphocytes and rat spleen mononuclear cells were compared with Homologous rat mononuclear cells were mixed culture, and the levels of cell proliferation were measured by liquid scintillation method. Results The proliferation of Foxp3 group (3 354.18 ± 203.73) was slightly higher than that of the control T cell group (2919.25 ± 137.91), but significantly lower than that of the blank control group (12 706.06 ± 967.47). Conclusion Retrovirus-mediated Foxp3 gene can be highly expressed on CD4 + CD25-T lymphocytes, and make the cells acquire immune incompetence and immunosuppressive properties similar to those of CD4 + CD25 + T lymphocytes.