成熟树突状细胞和转化生长因子β联合体外扩增小鼠调节性T细胞

背景:目前天然CD4+CD25+Foxp3+调节性T细胞稀少,远远不能满足临床需求。目的:建立有效的体外培养扩增小鼠CD4+CD25+T细胞体系。方法:从C57/BL6小鼠骨髓获得的成熟树突状细胞联合不同剂量转化生长因子β,体外扩增同基因或异基因CD4+CD25+T细胞。结果与结论:经过2周培养,磁珠分选后的CD4+CD25+T细胞的扩增倍数为自然调节性T细胞的(17.3±10.6)倍,0.2,2μg/L质量浓度对T细胞的扩增能力最强;经扩增后的T双阳性率为(85.38±1.82)%,CD127阳性率(78.86±0.91)%;体外扩增的调节性T细胞高表达FOXP3为天然调节性T细胞的1.5倍;调节性T细胞体外抑制实验结果表明:调节性T细胞对自体或异体CD4+T细胞抑制能力随着效靶比浓度的降低而降低,体外扩增的CD4+CD25+T细胞对自体CD4+T细胞抑制率(60.1±0.71)%,对异体CD4+T抑制率为(35.5±0.57)%(P<0.05)。提示在体外成功扩增了CD4+CD25+T细胞,并且具有免疫抑制功能,此方法扩增细胞数量多,细胞纯度有很大提高,方法简便。

Combination of mature dendritic cells and transforming growth factor beta for in vitro amplification of regulatory T cells in mice

BACKGROUND:At present,the nature CD4+CD25+Foxp3+ regulatory T cells are too rare to meet the clinical requirement.OBJECTIVE:To establish an effective amplification system of murine CD4+CD25+T cells in vitro.METHODS:Mature dendritic cells from C57/BL6 mouse bone marrow were combined with different doses of transforming growth factor β were used to amplify autologous or allogeneic CD4+CD25+T cells in vitro.RESULTS AND CONCLUSION:After 2 weeks culture,magnetic bead sorted CD4+CD25+T cells in the amplification factor for natural regulatory T cells were(17.3±10.6) times,0.2 and 2 μg/L concentration had the strongest amplification on T cells.The expansion CD4+CD25+ positive rate was(85.38±1.82)%,CD127 positive rate was(78.86±0.91)%;The expansion of regulatory T cells with high expression of FOXP3 mRNA was 1.5 folds than nature ones in vitro.The regulatory T cells inhibitory test showed that regulatory T cells could inhibit the proliferation of auto-and allo-CD4+T cells in vitro in a cell dose-dependent manner.The inhibit rate of CD4+CD25+T cells to autologous CD4+T cells was(60.1±0.71) %,the inhibit rate of allogeneic CD4+T cells was(35.5±0.57)%(P < 0.05).We successfully amplified CD4+CD25+T cells in vitro.The cells expanded have inhibitory effects in vitro,and as the method can obtain large quantity and purity cells,which may be used in clinical trials in future.