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目的 研究细胞因子组合体外诱导急性髓系白血病 (AML)细胞分化为树突状细胞 (DC)的可行性及AML细胞衍生DC(AML DC)的生物学特性。方法 AML细胞分别在GM CSF +IL 4、GM CSF +TNF α或GM CSF +IL 4 +TNF α 3种细胞因子组合以及不含细胞因子的培养液中培养。通过细胞形态动态观察、细胞化学染色和细胞免疫表型鉴定DC。用混合淋巴细胞培养 (MLC)、FITC标记的葡聚糖摄入实验和LDH释放实验检测DC功能。RT PCR和FISH检测AML DC的特异融合基因。结果 15例AML细胞在 3种细胞因子作用下发生典型的DC形态变化。AML DC的DC相关表面分子CD1a、CD80 、CD86 、CD1 0 6 、CD83和HLA DR等较未培养或不加细胞因子培养的AML细胞表达明显上调 (P <0 0 5 )。AML DC的异基因刺激能力明显高于未培养或不加细胞因子培养的AML细胞 (P <0 .0 5 )。只有用GM CSF +IL 4培养的AML DC有吞噬能力。AML DC与未培养AML细胞致敏的疾病初发时分离的T细胞比较 ,对自体AML细胞无明显的杀伤活性。AML DC仍具有未培养AML细胞的特异融合基因。结论 体外细胞因子可诱导各型AML细胞分化为DC ,细胞因子组合不同 ,AML DC的成熟状态可有一定的差异。AML DC不但起源于AML细胞 ,而且具有正常DC的典型形态、表型和功能。AML细胞可导致自体T细胞失能
Objective To investigate the feasibility of differentiation of acute myeloid leukemia (AML) cells into dendritic cells (DC) induced by cytokines in vitro and the biological characteristics of AML DCs. Methods AML cells were cultured in a combination of three cytokines, GM CSF + IL 4, GM CSF + TNFα, or GM CSF + IL 4 + TNFα, and cytokine-free media. DCs were identified by dynamic observation of cell morphology, cytochemical staining and cellular immunophenotype. DC function was tested by mixed lymphocyte culture (MLC), FITC-labeled dextran uptake assay and LDH release assay. The specific fusion gene of AML DC was detected by RT PCR and FISH. Results 15 cases of AML cells under the action of three kinds of cytokines typical DC morphological changes. The expression of DC-related surface molecules CD1a, CD80, CD86, CD1 0 6, CD83 and HLA DR in AML DCs were significantly up-regulated compared with those in non-cultured or untreated AML cells (P <0.05). Allogeneic stimulation of AML DC was significantly higher than that of AML cells cultured with or without cytokines (P <0.05). Only AML DCs cultured with GM CSF + IL4 have phagocytosis. AML DCs showed no cytotoxic activity on autologous AML cells when compared to T cells isolated on the first episode of a disease sensitized with no AML cells. AML DCs still have a specific fusion gene that is not cultured AML cells. Conclusion In vitro cytokines can induce the differentiation of various types of AML cells into DCs, and the combination of cytokines is different. The maturation status of AML DCs may have some differences. AML DCs originate not only from AML cells but also from the typical morphology, phenotype and function of normal DCs. AML cells can cause autologous T cell disability