hIL-10基因修饰骨髓间充质干细胞对异种T淋巴细胞增殖的影响及机制研究

目的 研究hIL-10基因修饰的MSCs(骨髓间充质干细胞)对异种混合T淋巴细胞增殖的影响及机制.方法 RT-PCR克隆hIL-10基因并将构建慢病毒hIL-10/pLOX-cwGFPs,然后将慢病毒转导hIL-10入豚鼠MSCs;ELISA测定hIL-10-MSCs培养上清hIL-10含量;CCK-8法检测hIL-10基因修饰的骨髓间充质干细胞对体外混合淋巴细胞培养的影响;ELISA测定其培养上清对外周血单个核细胞分泌IFN-γ的影响.结果 成功构建慢病毒hIL-10/pLOX-cwGFP,hIL-10-MSCs培养上清的IL-10水平明显高于未转染(P<0.05);hIL-10-MSCs能抑制异种T淋巴细胞混合培养的增殖反应(P<0.05);加入IL-2能逆转MSCs的抑制作用(P<0.05);hIL-10-MSCs培养上清组能抑制PHA刺激的PBMC分泌IFN-γ(P<0.05).结论 hIL-10-MSCs对混合异种淋巴细胞培养有显著的抑制作用,能抑制PHA刺激的PBMC分泌IFN-γ.

An experimental study on role of hIL-10-MSCs in heterologous T cell proliferation

Objective To observe the effects of hIL-10-MSCs on heterologous T cell prolifera-tion. Methods hlL-10 was cloned by RT-PCR and then hIL-10/pLOX-cwGFPs was construct. The lentivirus was transfected into cavy MSCs and cell culture supernatant was tested for IL-10 protein by ELISA assay. The effects of hIL-10-MSCs on heterologous T cell proliferation were determined by CCK-8. The effect of peripheral blood mononuclearcell secretion IFN-γ was tested by EIASA. Results hIL-10/pLOX-cwGFP were constructed successfully and hIL-10-MSCs cell culture supernatant of IL-10 protein were increased obviously, hIL-10-MSCs could inhibit heterologous T cell proliferation sig-nificantly(P<0.05) but could not induce T cell proliferation (P<0.05). Adding IL-2 could reverse this inhibition(P<0.05), hIL-10-MSCs cell culture supernatant could inhibit peripheral blood mono-nuclearcell secretion IFN-γ. Conclusion hIL-10-MSCs may play an important role in significant im-munosuppression of heterologous T cell proliferation and suppression of secretion IFN-γ by peripheral blood mononuclear cells.