急性髓细胞白血病细胞诱导分化成为树突状细胞的实验研究

The study of differentiation of acute myeloid leukemic cells into dendritic cells

目的 :探讨急性髓细胞白血病 (AML)患者的白血病细胞体外是否可诱导分化成为树突状细胞(DC)。方法 :从 11例AML患者的骨髓或外周血中获取非贴壁细胞 ,利用细胞因子 (rhGM CSF、rhIL 4、rhTNF α)联合培养 ,每 12d收获细胞。培养前后分别用倒置显微镜、电镜观察细胞形态 ,用流式细胞术测定细胞表面标志 ,用MTT法检测收获细胞激发混合淋巴细胞反应的能力。结果 :8例AML标本分化成为具有典型树突状形态的细胞 ,培养后的白血病细胞的HLA DR、CD86、CD1a表达较培养前明显增高 ,差异有统计学意义 (P <0 .0 1)。在混合淋巴细胞反应中 ,DC具有强烈激发同种T淋巴细胞增殖的能力 ,且随数量增加而作用增强。M4/M5型AML DC的表面标志的表达率明显高于非M4/M5型AML DC(P <0 .0 1)。结论 :急性髓细胞白血病细胞可以诱导分化成为具有DC形态、表型、功能的细胞。

Objective:To investigate whether dendritic cells could be generated from leukemic cells of patients with acute myeloid leukemia.Method:Nonadherent cells from bone marrow or peripheral blood mononuclear cells of 11 patients with AML were cultured with rhGM-CSF, rhIL-4 and rhTNF-α. All the cells were harvested at the twelfth day.The morphology of cells were detected by microscope, invert microscope and electron microscope, while the phenotypes were analyzed by flow cytometry. The proliferation of T cells was performed by methyl thiazolyl tetrazolium (MTT) assay.Result:After culture, cells from 8/11 samples exhibited typical morphological features of DCs. The expression of HLA-DR, CD86 and CD1a were much higher than those before culture. There were significantly statistical difference among them(P< 0.01). These AML-DCs had more powerful capacity than that of control in mixed lymphocyte responses, and the function was increased as the number of DC was increased. The expression of these three kinds of markers of M 4/M 5 AML-DC were much higher than those of other kinds of AML types(P< 0.01).Conclusion:AML cells can differentiate into dendritic like cells which exhibited DC's morphology, phenotypes and functions.