Ning Hong-Mei, Jin Jian-Gang, Hu Jiang-Wei, Feng Kai, Chen Hu
Department of Hematopoietic Stem Cell Transplantation, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100039, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Feb;13(1):43-9.
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.
目的是研究间充质干细胞（MSC）对免疫功能的影响，并探索在异基因骨髓移植中预防移植物抗宿主病（GVHD）和宿主抗移植物反应（HVGR）的新策略。从人骨髓细胞中分离并培养MSC。通过显微摄影用纺锤形纤维母细胞形态特征鉴定MSC的纯度，并用流式细胞术检测其表型。将MSC接种于6孔板中（A组8×10⁴/孔，B组4×10⁴/孔，C组2×10⁴/孔），并与从外周血分离的T细胞共培养7天。未与MSC共培养的外周血T细胞作为对照组。分别在与MSC共培养0、24、72小时和7天后，通过流式细胞术分析T细胞上表达的CD3、CD4、CD8和CD25。结果显示，与对照组（未与MSC共培养的T淋巴细胞）相比，与骨髓MSC共培养的异基因T淋巴细胞（A组和B组）的CD4⁺CD25⁺T细胞和CD8⁺T细胞明显增加，A组和B组之间无差异。实验组和对照组T细胞上表达的CD3、CD4、CD8和CD25无显著差异。结果表明，与异基因MSC共培养时，CD4⁺CD25⁺调节性T细胞和CD8⁺T细胞明显增加。结论是，MSC预处理可能有助于预防异基因骨髓移植中的GVHD和HVGR，并为诱导移植耐受提供了一种新策略。
