Objective To observe the influence of tumor necrosis factor-α(TNF-α) on the level of dendritic cell-chemokine l(DC-CKl) expressed by dendritic cells (DCs) in vitro induced from chronic myeloid leukemia (CML). Methods Peripheral blood monouclear cells (PBMC) were isolated from 20 newly diagnosed CML patients, and co-cultured with recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) 100 μg/L and recombinant human interleukin-4(rhIL-4) 100 μg/L for 5 days,then TNF-α 50 μg/L was added to experimental group, control group only contained medium, the cells were harvested at the day 7th. DCs were studied by cell morphology (Wright staining, inverted microscope) and immunophenotype; fluorescence in situ hybridization (FISH) was used to analyze the cytogenetics. The level of DC-CK1 was measured by enzyme-linked immunosorbent assay(ELISA). Results Dendrite-like projection of experimental group was more obvious than that of control group, and the expression of surface marker in experimental group was more up-regulated than that of control group, the difference had statistical significance( P <0. 05). Meanwhile, the origin of leukemia was confirmed by FISH. ELISA showed that the level of DC-CK1 expressed by experimental group was (175. 094±52. 487) ng/L( n =10),which was lower significantly than that of control group (658. 926 + 84. 105) ng/L( n = 10), and the difference had statistical significance( P <0. 05). Conclusion TNF-α inhibited the generation of DC-CK1 expressed by CML-DCs.%目的 观察肿瘤坏死因子a(TNF-α)对体外诱导的慢性粒细胞白血病(CML)来源的树突状细胞(DCs)表达树突状细胞趋化因子1(DC-CK1)水平的影响.方法分离初诊20例CML患者外周血单个核细胞,用重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)100ug/L、重组人白细胞介素4(rhIL-4)100μg/L培养5天,然后分为实验组和对照组两组,实验组加入TNF-a 50μg/L,对照组只加培养基,第7天收获细胞.通过细胞形态学(倒置显微镜、Wright染色)及免疫表型对培养的DCs进行鉴定；荧光原位杂交(FISH)对DCs进行细胞遗传学检测；酶联免疫吸附测定(ELISA)法检测DC-CK1的表达水平.结果培养后实验组细胞的树枝样突起更明显,而且实验组DCs表面分子的表达比对照组上调,差异有统计学意义(P＜0.05),同时经FISH证实了DCs的白血病来源；最后ELISA检测显示实验组DCs培养上清中DC-CK1的含量为(175.094±52.487)ng/L(n=10),明显低于对照组的(658.926±84.105)ng/L(n=10),差异有统计学意义(P＜0.05).结论TNF-α抑制CML源树突状细胞表达DC-CK1.
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