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目的体外观察妊娠浓度的雌激素能否诱导CD4+CD25-naive T细胞转化为CD4+CD25+Treg细胞,并探讨其相关性。方法以CD4+CD25-T细胞作为反应细胞,实验组加入妊娠水平的雌激素(E2)及CD3/CD28单抗作为刺激原培养72 h,设阴性对照组(仅加入CD3/CD28单抗)和空白对照组。72 h后检测各组中CD4+CD25+T细胞和CD4+Foxp3+T细胞比例变化及Foxp3 mRNA表达。结果1)阴性对照组CD4+CD25+T细胞比例较空白对照组显著增高(P<0.001),而实验组CD4+CD25+T细胞比例进一步升高(P<0.001)。2)阴性对照组不能诱导CD4+Foxp3+T细胞比例增高,但实验组CD4+Foxp3+T细胞的比例则较其它2组均显著升高(P<0.001)。3)RT-PCR提示阴性对照组Foxp3 mRNA的表达量较空白对照组无显著差异(P>0.05);而实验组Foxp3 mRNA的表达量较其它2组均显著升高(P<0.001)。结论体外淋巴细胞刺激实验提示妊娠状态下雌激素的高水平与CD4+CD25+Foxp3+Treg细胞比例的升高密切相关。
Objective To observe whether estrogen at gestational levels can induce the conversion of CD4 + CD25-naive T cells to CD4 + CD25 + Treg cells in vitro and to investigate their correlation. Methods CD4 (superscript +) CD25 (superscript +) T cells were used as reactive cells. Pregnancy-induced estrogen (E2) and CD3 / CD28 monoclonal antibodies were used as primary cultured cells for 72 h. Negative control group Blank control group. After 72 hours, the proportion of CD4 + CD25 + T cells and CD4 + Foxp3 + T cells and the expression of Foxp3 mRNA in each group were detected. Results 1) The proportion of CD4 + CD25 + T cells in the negative control group was significantly higher than that in the blank control group (P <0.001), while the proportion of CD4 + CD25 + T cells in the experimental group was further increased (P <0.001). 2) The negative control group could not induce an increase in the proportion of CD4 + Foxp3 + T cells, but the proportion of CD4 + Foxp3 + T cells in the experimental group was significantly higher than the other two groups (P <0.001). RT-PCR indicated that the expression of Foxp3 mRNA in the negative control group was not significantly different from that in the blank control group (P> 0.05), while the expression of Foxp3 mRNA in the experimental group was significantly higher than the other two groups (P <0.001). Conclusion In vitro lymphocyte stimulation experiments suggest that the high level of estrogen in pregnancy is closely related to the increased proportion of CD4 + CD25 + Foxp3 + Treg cells.