摘要
目的研究hII-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hII-10人豚鼠MSCs；ELlSA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-Y的影响。结果成功构建慢病毒hIL-10／pLOX-ewGFP；hIL-10-MSCs培养上清的IL-10水平明显高于未转染（P〈O．05）；hlL-10-MSCs能抑制异种T淋巴细胞混合培养的增殖反应（P〈O．05）；加入IL-2能逆转MSCs的抑制作用（P〈O．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。
Objective To observe the effects of hIL-10-MSCs on heterologous T cell prolifera tion. Methods hIL-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 hlL-10-MSCs on heterologous T cell proliferation were determined by CCK-8. The effect of peripheral blood mononuclearcelI secretion IFN-γ was tested by ELISA. 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）, hlL-10-MSCs cell culture supernatant could inhibit peripheral blood mono- nucleareell secretion IFN-7. 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.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2010年第2期138-141,共4页
Chinese Journal of Hepatobiliary Surgery
基金
基金项目：国家自然科学基金项目资助（30801125）
