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    TGF-β1诱导的新生儿脐带血CD4+CD25+调节性T细胞增殖动力学研究

    The dynamics study of proliferation of CD4+CD25+ Treg cells induced by transforming growth factor-β1 in neonatal cord blood

      摘要
    • 摘要: 目的:探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的新生儿脐带血CD4+CD25+调节性T细胞增殖活性。方法:新生儿脐带血单个核细胞经羧基荧光素乙酰乙酸琥珀酰亚胺酯(carboxyfluorescein diacetate,succinimidyl ester,CFSE)标记后,加入TGF-β1,同时用CD3mAb激活和IL-2培养扩增。培养第4、6、8天收集细胞,用流式细胞术检测活化增殖后的CD4+CD25+、CD8+ T细胞增殖动力学变化。结果:培养后第6、8天,实验组CD4+CD25+ T细胞增殖指数分别为11.52、23.04,高于对照组CD4+CD25+ T细胞(6.68,10.46)及实验组CD8+ T细胞(4.23,5.43)。培养后第8天,实验组CD4+CD25+ T细胞第8代到第10代所占的比例为52.74%,在各代中以第8代所占比例最高,而对照组CD4+CD25+ T细胞第8代到第10代所占的比例为36.26%;实验组CD8+ T细胞第8代到第10代所占的比例为13.54%,在各代中所占比例最高的为第2代,而对照组CD8+ T细胞第8代到第10代所占的比例为29.44%。结论:TGF-β1诱导的新生儿脐带血CD4+CD25+调节性T细胞增殖活性明显强于CD8+ T细胞。

       

      Abstract: Objective: To explore proliferative activity of CD4+CD25+ Treg cells induced by transforming growth factor-β1(TGF-β1) in neonatal cord blood.Methods: Cord blood mononuclear cells were labeled with carboxyfluorescein diacetate,succinimidyl ester(CFSE),and then stimulated with TGF-β1,CD3mAb and cultured in 10% NBS-1640 medium in presence of IL-2.The dynamics of proliferation of CD4+CD25+ and CD8+ T cells on day 4,day 6,day 8 after culture was determined by flow cytometry.Results: Proliferative kinetics analysis showed proliferation index(PI) on day 6 and day 8 of culture with TGF-β1 in CD4+CD25+ Treg cells(11.52 and 23.04) were significantly higher than those in control cells(6.68 and 10.46) and those cultured with TGF-β1 in CD8+ T cells(4.23 and 5.43).On day 8 of culture with TGF-β1,proportion of the generations from 8th to 10th in CD4+CD25+ Treg(52.74%) was strikingly higher than those in control cells(36.26%) and those in CD8+ T cells cultured with TGF-β1(13.54%),while the proportion of the generations from 8th to 10th in control CD8+ T cells was 29.44%.Conclusions: Proliferative activity of CD4+CD25+ Treg cells induced by TGF-β1 in neonatal cord blood was obviously stronger than that of CD8+ T cells.

       

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