骨髓间充质干细胞的体外扩增及其对T淋巴细胞产生IFN-γ和IL-10的影响

In vitro proliferation of mesenchymal stem cells and its effect on the production of IFN-γ and IL-10 by T lymphocytes

为了探讨骨髓间充质干细胞(mesenchymal stem cells,MSC)对T淋巴细胞分泌功能的免疫调节作用,从人骨髓分离培养MSC,通过其形态的均一性及流式细胞术检测表面标志以鉴定其纯度;从外周血分离获得T淋巴细胞,再将MSC分别以不同数量加入到植物血凝素(PHA)刺激的外周血T淋巴细胞和混合淋巴细胞反应(MLR)培养体系及不同浓度MSC培养上清加入混合淋巴细胞反应培养体系共培养后,分别收集上清,ELISA检测IFN-γ和IL-10水平,发现不同细胞数量的MSC对T细胞分泌细胞因子IFN-γ均有抑制,同时促进IL-10分泌,且其抑制和促进均呈剂量依赖性;MSC的不同上清浓度对IFN-γ抑制亦呈浓度依赖性,但未发现对IL-10分泌的影响。表明骨髓MSC在体外可抑制T细胞分泌IFN-γ、促进分泌IL-10,起到免疫调节作用。

To investigate the immuno-regulatory effect of mesenchymal stem cells（MSC） on the production of IFN-γ and IL-10 by T lymphocytes, MSC isolated from human bone marrow cells were cultivated and their purity was identified by the homogenicity with a spindle shaped fibroblastic morphology and by their phenotypes tested by flow cytometry. The effects of different amounts of MSC（5 000, 10 000, 40 000 cells/well） on the secretory function of the PHA- activated T lymphocytes（group A） and on the secretory function of T lymphocytes as determined by MLR（group B）, as well as the effect of different concentrations of the MSC-cultural supernatants（group C） were determined accordingly,by the testing of secretion of IFN-γ and IL-10 in the supernatants of MSC-culturings with ELISA assay. It was demonstrated that the effect of different amounts of MSC on the secretion of IFN-γ was found to be inhibitory,but they could promote the secretion of IL-10 ,while the inhibitory and the enhancing effects were dose-dependent. The inhibitory effect of cultural supernatants of MSC on the secretion of IFN-γ by T cells was concentration-dependent,but no any effect could be found on the secretion of IL-10. These results indicate that MSC can induce the inhibition of IFN-γ secretion but promote the secretion IL-10 by T lymphoeytes in vitro to mediate a immuno-regula-tory effect of T lymphocytes.