人骨髓间充质干细胞体外对异基因T淋巴细胞表型的影响

为了研究间充质干细胞 (mesenchymalstemcell,MSC)的免疫调节作用 ,探讨防治异基因骨髓移植中移植物抗宿主病 (GVHD)和移植排斥反应 (HVGR)的可能途径 ,从人骨髓中分离培养间充质干细胞 ,并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 ,将MSC分别以 8× 10 4 (A组 )、4× 10 4 (B组 )和 2× 10 4 (C组 )个细胞 孔分别接种于 6孔板 ,并与经分离的外周血T淋巴细胞共培养 7天 ,以单独培养的外周血T淋巴细胞为对照组 ,用流式细胞仪分别在培养 0、2 4、72小时和 7天测定各组T细胞上CD3、CD4、CD8、CD2 5表达的变化。结果表明 :T细胞和MSC共培养组与T细胞单独培养组相比 ,A组和B组CD4 CD2 5 免疫调节性T细胞和CD8 T细胞明显增多 ,A组和B组之间无明显差别。实验组和对照组相比CD3、CD4、CD2 5表达无明显差别。结论 :骨髓MSCs在体外可使外周血T细胞表型发生改变 ,CD4 CD2 5 免疫调节性T细胞和CD8 T细胞明显增多 ,本研究结果为临床异基因造血干细胞移植预防GVHD时MSCs的输注数量提供了参考。

Effect of Human Bone Marrow Mesenchymal Stem Cell on Allogeneic T Lymphocyte Phenotype in vitro

The purpose was to study the effect of mesenchymal stem cell (MSC) on immune function, and to explore the new strategy to prevent graft versus host disease (GVHD) and host versus graft reaction (HVGR) in allogeneic bone marrow transplantation. MSCs from human bone marrow cells were isolated and cultured. The purity of MSCs were identified with the spindle-fibroblastic morphological characterization by microphotography, and the phenotypes were tested by flow cytometry. MSCs were planted in 6-well plates (8 x 10(4)/well for group A, 4 x 10(4)/well for group B and 2 x 10(4)/well for group C), and cocultured for 7 days with T cell isolated from peripheral blood. Peripheral blood T cells non-cocultured with MSC acted as the control group. CD3, CD4, CD8, and CD25 expressed on T cells were analyzed by flow cytometry after coculture with MSCs for 0, 24, 72 hours and 7 days respectively. The results showed that CD4(+)CD25(+) T cells and CD8(+) T cells of allogeneic T lymphocytes cocultured with bone marrow MSCs (group A and group B) increased obviously as compared with control group (T lymphocytes uncocultured with MSCs), and there were no difference between groups A and B. CD3, CD4, CD8 and CD25 expressed on T cells had no significant difference between experimental groups and control group. The result suggested that CD4(+)CD25(+)-regulatory T cells and CD8(+) T cells were increased apparently when cocultured with allogeneic MSCs. It is concluded that MSCs pretreatment may be useful in the prevention of GVHD and HVGR in allogeneic bone marrow transplantation and provides a new strategy to induce transplantation tolerance.