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目的:狼疮方对体外扩增小鼠CD4+CD25+调节性T细胞数量的影响。方法:免疫磁珠法分离狼疮方治疗组与未治疗组小鼠CD4+CD2+5T细胞;通过流式细胞仪检测CD4+CD25+T细胞Foxp3表达情况;用抗CD3和抗CD28包被的微珠来进行刺激,体外扩增小鼠的CD4+CD25+调节性T细胞;用CFSE法检测CD4+CD25+调节性T细胞体外抑制CD4+CD2-5T细胞的增殖情况。结果:经抗CD3和抗CD28包被的微珠刺激后,在TGF-β1和IL-2存在的环境中CD4+CD25+T细胞数量明显升高,扩增后的CD4+CD25+T细胞高表达Foxp3。与此同时,经狼疮方处理后的小鼠CD4+CD25+T细胞扩增后数量增加133.3±18.9倍,未经狼疮方处理的对照细胞数量增加104.7±10.8倍,两者比较有显著性差异(P<0.05)。狼疮方治疗组与未治疗组中分离的CD4+CD2+5T细胞和扩增的CD4+CD25+T细胞对CD4+CD2-5T细胞的增殖皆有明显抑制作用,荧光强度分别为32.1±4.8,33.1±4.9,30.1±3.8,32.9±4.2。结论:狼疮方有助于提高小鼠CD4+CD25+调节性T细胞的扩增效率,扩增的CD4+CD25+调节性T细胞表达Foxp3,具有体外免疫抑制功能。
Objective: To investigate the effect of lupus prescription on the number of CD4 + CD25 + regulatory T cells in vitro. Methods: CD4 + CD25 + T cells were isolated from mouse lupus group and untreated group by immunomagnetic beads method. The expression of Foxp3 in CD4 + CD25 + T cells was detected by flow cytometry. The anti-CD3 and anti- Beads to stimulate CD4 + CD25 + regulatory T cells in vitro expansion of mice; using CFSE assay CD4 + CD25 + regulatory T cells in vitro inhibition of CD4 CD2-5 T cell proliferation. Results: After stimulation with anti-CD3 and anti-CD28 beads, the number of CD4 + CD25 + T cells in the presence of TGF-β1 and IL-2 significantly increased, and the high CD4 + CD25 + T cells Foxp3 is expressed. At the same time, the number of mouse CD4 + CD25 + T cells treated with lupus increased by 133.3 ± 18.9 folds, while the number of control cells without lupus increased by 104.7 ± 10.8 folds, showing significant difference between the two (P <0.05). CD4 + CD25 + T cells and CD4 + CD25 + T cells isolated from lupus treatment group and untreated group had significant inhibitory effects on the proliferation of CD4 + CD25T cells with fluorescence intensities of 32.1 ± 4.8, 33.1 ± 4.9, 30.1 ± 3.8, 32.9 ± 4.2. Conclusion: Lupus prescription can improve the efficiency of mouse CD4 + CD25 + regulatory T cells, and the expanded CD4 + CD25 + regulatory T cells express Foxp3, which has the function of immunosuppression in vitro.