摘要
目的:观察干扰素-γ(IFN-γ)增强人脐带间充质干细胞(HUMSCs)在体外对CD4+T细胞增殖的抑制作用。方法:采用胶原酶Ⅱ对人脐带组织进行消化,分离出间充质干细胞(MSCs),贴壁培养,经过传代纯化后,在体外将HUMSCs与CD4+T细胞[健康成人的外周血单个核细胞(PBMC)用免疫磁珠分选得到]共培养,CD4+T细胞用羧基荧光素乙酰乙酸琥珀酰亚胺酯(CFSE)标记,流式细胞仪测定CD4+T细胞的增殖情况,并通过反转录-聚合酶链反应(RT-PCR)以及Westernblot分别测定未经处理的HUMSCs以及IFN-γ处理后HUMSCs的吲哚胺-2,3-双加氧酶(IDO)基因及蛋白表达水平。结果:IFN-γ处理后的HUMSCs能够抑制CD4+T细胞的增殖(P<0.05),而未处理的HUMSCs对CD4+T细胞的增殖抑制作用不显著。IFN-γ处理后的HUMSCs能够表达IDO,而未处理组则不表达IDO。结论:IFN-γ能够通过IDO的表达增强HUMSCs在体外对CD4+T细胞增殖的抑制作用。
Objective To investigate that IFN-γ can promote the immunosuppressive capacity of human umbilical cord mesenchymal stem cells （HUMSCs） by expression of indoleamine 2,3-dioxygenase （IDO） in vitro. Methods Human umbilical cord was digested by collagenase type Ⅱto obtain mesenchymal stem cells,and the cells were purified by adherent culture and passage. HUMSCs and CD4＋T cells were cocultured in vitro. The CD4＋T cells were obtained by magnetic activated cell sorting and labelled with CFSE （carboxyfluorescein diacetate succinimidyl ester）,and flow cytometry was used to measure the proliferation of CD4 ＋ T cells. RT-PCR and Western blot were used to measure IDO gene expression and protein levels in IFN-γ treated and untreated HUMSCs. Results IFN-γ treated HUMSCs had a strong inhibitory effect on the proliferation of CD4 ＋ T cells （P〈0.05）,yet untreated HUMSCs could not obviously inhibit the proliferation of CD4 ＋ T cells（P〉0.05）. IFN-γ treated HUMSCs were able to express IDO,whereas untreated HUMSCs did not express IDO. Conclusions IFN-γ can promote the immunosuppressive capacity of HUMSCs on proliferation of CD4 ＋ T cells through the expression of IDO.
出处
《诊断学理论与实践》
2010年第2期181-185,共5页
Journal of Diagnostics Concepts & Practice
