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目的探讨利用小鼠同种异型骨髓源性树突状细胞(BMDC)与CD4+T细胞共培养体外扩增CD4+CD25+调节性T(CD4+CD25+Treg)细胞的方法。方法将小鼠同种异型imBMDC与CD4+T细胞共培养,通过测定细胞增殖率和CD4+CD25+Treg细胞占CD4+T细胞比例的变化评定该方法体外扩增CD4+CD25+Treg细胞的能力;利用荧光定量RTPCR检测体外扩增CD4+CD25+Treg细胞FOXP3基因表达及细胞增殖抑制实验检测其细胞功能活性。结果共培养d 7细胞增殖率(5.26±0.286)倍;CD4+CD25+Treg细胞占CD4+T细胞的(33.77±3.69)%,较新鲜分离小鼠脾细胞中CD4+CD25+Treg细胞比例明显增加(P﹤0.05)。体外扩增CD4+CD25+Treg细胞FOXP3基因的表达与新鲜分离CD4+CD25+Treg细胞差异无统计学意义(P﹥0.05)。混合淋巴细胞培养证实体外扩增CD4+CD25+Treg细胞的细胞增殖抑制作用较新鲜分离的CD4+CD25+Treg细胞更强(P﹤0.05)。结论小鼠同种异型BMDC与CD4+T细胞共培养能有效诱导扩增CD4+CD25+Treg细胞并保持其免疫细胞功能活性。
Objective To explore a method of expanding CD4 + CD25 + regulatory T (CD4 + CD25 + Treg) cells in vitro by coculturing mouse allogenic bone marrow-derived dendritic cells (BMDC) and CD4 + T cells. Methods Mouse allogeneic imBMDC were co-cultured with CD4 + T cells. The ability of this method to expand CD4 + CD25 + Treg cells in vitro was evaluated by measuring the rate of cell proliferation and the ratio of CD4 + CD25 + Treg cells to CD4 + T cells The cell viability of FOXP3 gene was detected by fluorescent quantitative RTPCR in vitro. The results showed that the proliferation rate of cultured d 7 cells was (5.26 ± 0.286) times higher than that of the control group. The percentage of CD4 + CD25 + Treg cells in CD4 + T cells (33.77 ± 3.69)% was significantly higher than that in freshly isolated mouse spleen cells Increase (P <0.05). The FOXP3 gene expression in CD4 + CD25 + Treg cells expanded in vitro was not significantly different from that of freshly isolated CD4 + CD25 + Treg cells (P> 0.05). Mixed lymphocyte culture confirmed that in vitro expansion of CD4 + CD25 + Treg cell proliferation inhibition than freshly isolated CD4 + CD25 + Treg cells stronger (P <0.05). Conclusion The co-culture of mouse BMDC and CD4 + T cells can effectively induce the expansion of CD4 + CD25 + Treg cells and maintain their immune cell functional activity.