人骨髓间充质干细胞对同种异体调节性T细胞的影响及可能机制实验研究论文.docVIP

　　人骨髓间充质干细胞对同种异体调节性T细胞的影响及可能机制实验研究论文 杨靖，王庆海，曾秋棠，毛晓波 【摘要】 本研究探讨人骨髓间充质干细胞（human mesenchymal stem cells, hMSC）对同种异体CD4+CD25+调节T细胞的作用，进一步查明人骨髓间充质干细胞对T淋巴细胞免疫调节作用的可能机制。从人骨髓中分离培养hMSC，通过免疫组织化学方法检测其表面标记并进行鉴定。从非亲缘供者健康人外周血中提取单个核细胞，在IL2作用下体外培养， 实验组加入不同数量级hMSC（1×103.freelinute, CPM值），细胞因子TGFβ、 IL10、IFNγ和IL12表达量的相关性。结果表明：hMSC能明显抑制T淋巴细胞增殖，且抑制作用与hMSC呈剂量依赖性；实验组各组CD4+CD25+T细胞亚群含量以及Foxp3相对表达水平均较对照组明显增加(p 0.05)，且实验组间比较差异仍具有统计学意义(p 0.05)；Foxp3表达水平、CD4+CD25+T细胞百分率与T淋巴细胞增殖水平呈明显负相关(p 0.05)，与TGFβ、IL10表达量呈明显正相关，而与IFNγ和IL12的表达无明显相关性。 结论：hMSC呈剂量依赖性抑制同种异体淋巴细胞增殖, 其机制可能与hMSC通过分泌TGFβ、IL10促进CD4+CD25-T细胞向表达Foxp3的CD4+CD25+调节T细胞转化，进而发挥免疫抑制作用有关。 【关键词】 骨髓间质干细胞 Effect of Human Bone Marroal Stem Cells on Allogeneic Regulatory T Cells and Its Possible Mechamism Abstract The study mune regulatory effects of human bone marroesenchymal stem cells (hMSCs) on Foxp3 expressing CD4+CD25+ regulatory T cells and to explore the mechanism of immune modulation by hMSCs. Human MSCs bone marroorphology, and the phenotypes munohistochemistry. Human peripheral blood mononuclear cells (hPBMNCs) an unrelated donor in the presence of rhIL2. After 5 days of coculture, the percentage of CD4+CD25+T cells etry. T cell proliferation idine incorporation using a liquid scintillation counter. The expression of Foxp3 in CD4+CD25+T cells erase chain reaction (RTPCR). Cytokines (TGFβ、IL12、IFNγ、IL10) concertrations of cultured supernatants easured ents, the presence of hMSCs anner. The increase of percentage of CD4+CD25+T cells in the peripheral CD4+ T cell phocytes RNA (Foxp3/βactin) in hMSCs groups RNA expression and the percentage of CD4+CD25+T cells. HoRNA expression and the percentage of CD4+CD25+T cells. It is concluded that the Foxp3 expressing regulatory T cells may play an important role in the immune regulatory by hMSCs. Its mechanism is related to that the hMSCsmediated TGFβ and IL10 convert CD4+CD25-T cells into CD4+CD25+T regulatory T cells, arroesenchymal stem cell; CD4+CD25+ T cell; Foxp3; imm