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本研究观察来源于慢性髓性白血病患者的CD34+细胞经两步培养扩增后产生的树突状细胞的抗白血病 免疫功能。从慢性髓性白血病患者骨髓或外周血中分离出CD34+细胞或外周血单个核细胞分别进行两步培养。 首先将其与rhFlt3-L和rhTPO共育7天,再与rhGM-CSF、rhTNF-α及rhIL-4共同培养14天,以诱导树突状细胞产 生:同时以rhGM-CSF、rhTNF-α及rhIL-4直接诱导体系作对照。获得的DC通过流式细胞仪检测免疫表型,电镜分 析细胞超微结构及G显带法进行的染色体分析后,并用MTT法检测DC刺激T细胞增殖能力及T细胞对CML细 胞的杀伤作用.结果表明:慢性髓性白血病的CD34+细胞经过rhFlt3-L和rhTPO的初始扩增培养7天后,CD34+细 胞总数扩增77±5倍。用rhGM-CSF、rhTNF-α及rhIL-4诱导培养14天,DC产率达到(39±8)％,细胞扩增倍数及 DC比例均明显高于直接诱导产生的培养方法(P<0.01),且此DC能刺激自体或异体T细胞增殖,进而杀伤CML 细胞。结论:两步法体外扩增培养可明显提高DC前体总数及产率,优于直接诱导培养;CML细胞来源的DC可诱 导产生抗白血病免疫反应。
This study was to observe the anti-leukemia immune function of dendritic cells derived from CD34 + cells from patients with chronic myelogenous leukemia after two-step culture. CD34 + cells or peripheral blood mononuclear cells were isolated from the bone marrow or peripheral blood of patients with chronic myelogenous leukemia and cultured in two steps separately. First, they were co-cultured with rhFlt3-L and rhTPO for 7 days and then cultured with rhGM-CSF, rhTNF-α and rhIL-4 for 14 days to induce dendritic cell production. The rhGM-CSF, rhTNF- rhIL-4 direct induction system as a control. DC obtained by immunocytochemistry by flow cytometry, electron microscopy analysis of cell ultrastructure and G banding method for chromosome analysis and MTT assay DC stimulation of T cell proliferation and T cell killing of CML cells. The results showed that the total number of CD34 + cells in chronic myelogenous leukemia CD34 + cells was 77 ± 5 times after the initial expansion of rhFlt3-L and rhTPO for 7 days. After cultured with rhGM-CSF, rhTNF-α and rhIL-4 for 14 days, the yield of DCs reached (39 ± 8)%, and the cell expansion ratio and DC ratio were significantly higher than those induced by rhGM- , And this DC can stimulate autologous or allogeneic T cell proliferation, thereby killing CML cells. Conclusion: The two-step in vitro culture can significantly increase the total number of DC precursors and yield, which is better than direct induction culture. CML-derived DC can induce anti-leukemia immune response.