【作者】 林梅；

【导师】 白海； 王存邦；

【作者基本信息】 兰州大学 ， 内科血液病学， 2008， 硕士

【摘要】 目的本实验探讨抑制基质细胞衍生因子1(SDF-1)的活性后,骨髓间充质干细胞(MSCs)培养上清对PHA刺激的异体T淋巴细胞免疫调节作用及急性髓性白血病细胞HL-60增殖的影响。方法应用Ficoll密度梯度离心法分离骨髓间充质干细胞,流式细胞术检测其表面标志。应用尼龙棉柱法分离异体外周血T淋巴细胞,常规培养HL-60细胞,并应用SDF-1受体CXCR4的单克隆抗体12G5阻断SDF-1的生物学效应。将MSCs上清与PHA刺激的T淋巴细胞及HL-60细胞共培养,应用MTT法检测细胞在应用单克隆抗体12G5前后增殖情况,并用ELISA法检测T淋巴细胞分泌的IL-4、IFN-γ水平。结果加入单克隆抗体12G5阻断SDF-1的生物学效应后,MSCs上清的促白血病细胞HL-60增殖的作用受到一定的抑制,MSCs上清抑制T淋巴细胞增殖的作用无明显的影响。T细胞单独培养组、T细胞与MSCs上清混合培养组、加入单抗12G5组培养上清测得IL-4的含量分别为8.27±2.68pg/ml、22.84±5.61pg/ml、5.35±2.51pg/ml,INF-γ的含量分别为11.75±4.95 pg/ml、165.81±14.73 pg/ml、186.99±45.09pg/ml。结论抑制MSCs分泌的SDF-1活性后,白血病细胞HL-60的增殖受到抑制,同时对T淋巴细胞增殖及T淋巴细胞分泌IFN-γ无明显影响,而T淋巴细胞分泌IL-4降低。更多还原

【Abstract】 ObjectiveTo investigate the immunoregulation of MSCs supernatant on proliferation of allogeneic T lymphocytes activated by PHA and Proliferation of Acute Myelocytic Leukemia Cell line HL-60 when the effects of SDF-1 were inhibited.MethodsBone marrow mesenchymal stem cells were isolated by Ficoll-Hypaque density gradient, and the surface marks were detected by flow cytometry. Allogeneic T lymphocytes from peripheral blood were harvested by nylon column and HL-60 cells were conventional cultured. SDF-1 activity was blocked with anti-CXCR4 McAb 12G5. Supernatant of MSCs was co-cultured with T lymphocytes activated by PHA and HL-60 cells. Proliferation of cells were detected by MTT. The secretion of the cytokines IL-4 and INF-γwere evaluated by ELISA.ResultsAfter SDF-1 activity was blocked with anti-CXCR4 McAb 12G5, the enhancement of MSCs supernatant on HL-60 cells was inhibited and the inhibition of MSCs supernatant on T lymphocytes was not obviously effected. The secretion of IL-4 were 8.27±2.68pg/ml、22.84±5.61pg/ml、5.35±2.51pg/ml and the secretion of INF-γwere 11.75±4.95 pg/ml、165.81±14.73 pg/ml、186.99±45.09pg/ml in the T-cells alone group, T-cells and MSCs supernatant co-culture group and adding McAb 12G5 group respectively.Conclusion Blocking SDF-1 activity can inhibit proliferation of leukemia cells and has no obvious effect on proliferation of allogeneic T Lymphocytes inhibited by MSCs supernatant and secretion of IFN-γ. The secretion of IL-4 was decreased.更多还原