慢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 CHRONIC MYELOID LEUKEMIC CELLS INDUCED TO DIFFERENTIATE INTO DENDRITIC CELLS

目的:将慢性髓性白血病患者的白血病细胞体外诱导分化成为树突状细胞。方法:从8例慢性髓性白血病患者的骨髓或外周血中获取非贴壁细胞,利用细胞因子(rhGM蛳CSF、rhIL蛳4、rhTNF蛳α)联合培养,每3 d补充新的细胞因子,其中rhTNF蛳α为第9天添加,第12天收获细胞。培养前后分别用光镜、倒置显微镜、电镜观察细胞形态,流式细胞仪测定细胞表面标志,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CML标本分化成为具有典型树突状形态的细胞,流式细胞仪检测白血病细胞的HLA蛳DR由(16.16±10.70)%增至(39.94±20.79)%,CD86由(3.53±1.75)%增至(8.83±6.11)%,CD1a由(8.78±4.90)%增至(19.17±8.51)%,差异有统计学意义(P <0.01)。在混合淋巴细胞反应中具有强烈激发同种T淋巴细胞增生的能力。结论:慢性髓性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 induce and differentiate into dendritic cells from leukemic cells of patients with chronic myeloid leukemia. Methods: Nonadherent cells from bone marrow or peripheral blood mononuclear cells of 8 patients with chronic myeloid leukemia were cultured with rhGM- CSF and rhIL- 4. The cytokines were supplemented every three days. RhTNF- α was used at the ninth day. All the cells were harvested at the twelfth day. The morphology of cells were detected by using microscope, invert microscope and electron microscope, while the phenotypes were analyzed by using flow cytometry. The proliferation of T cells was performed by methyl thiazolyl tetrazolium (MTT) assay. Result: After culture, cells from all the 8 samples exhibited typical morphogical features of DCs. Through flow cytometry , we found the expression of HLA- DR was increased from(16.16±10.70)% to(39.94±20.79)%, CD86 was increased from (3.53±1.75)% to (8.83±6.11)%, CD1a was increased from (8.78±4.90)% to(19.17±8.51)%. There were significantly statistical difference among them(P <0.01). These CML- DCs had more powerful capacity than that of control in mixed lymphocyte responses. Conclusion: Chronic myeloid leukemic cells can differentiate into dendritic like cells which exhibited DC's morphology, phenotypes and functions. These findings might prove useful for immunotherapy of CML.