目的： 研究树突状细胞（dendritic cell，DC）联合同源细胞因子诱导的杀伤细胞（cytokine-induced killer cell，CIK）对急性髓细胞白血病细胞株KG-1a中白血病干细胞（leukemic stem cell，LSC）的体外杀伤和诱导凋亡作用。 方法： 分离健康人外周血单个核细胞，贴壁细胞用GM-CSF和IL-4诱导培养DC，悬浮细胞用IL-2、IL-1、IFN-γ和CD3 mAb诱导培养CIK。将KG-1a细胞冻融物作为抗原负载DC（即Ag-DC），与CIK共培养作为实验组（Ag-DC-CIK），无抗原负载的DC与CIK共培养作为对照组（DC-CIK），单独CIK作为空白对照组，与KG-1a共育后流式细胞术检测各组细胞中CD34+CD38-CD123+白血病干细胞的比例。DC-CIK与KG-1a细胞共培养，流式细胞术检测各组细胞中KG-1a细胞与CD34+CD38-CD123+细胞的凋亡率。 结果： 外周血单个核细胞成功诱导DC。CIK组、DC-CIK组及Ag-DC-CIK组中CD3+CD56+细胞比例为（17.36±4.44）%、（28.22±3.66）%和（36.16±5.88）%，依次升高（P<0.05）。与对照组相比，Ag-DC-CIK组与DC-CIK组细胞中CD34+CD38-CD123+细胞比例显著降低\[（8.78±0.62）% vs（3.95±0.53）%、（3.03±0.62）%，P<0.01]。DC-CIK可诱导KG-1a细胞凋亡，凋亡率由（2.34±0.74）%上升至（12.27±1.01）%，但对其中CD34+CD38-CD123+细胞无明显的诱导凋亡作用。 结论： DC联合CIK能杀伤急性髓细胞白血病干细胞，但无明显的诱导凋亡作用。

Objective : To investigate the cytotoxicity effect of homologous cytokine-induced killer cells (CIKs), which was co-cultured with dendritic cells (DCs), against leukemic stem cells (LSCs) of acute myelogenous leukemic KG-1a cells. Methods: The peripheral blood mononuclear cells were isolated from healthy donors, and the adherent cells were induced to differentiate into DCs with GM-CSF and IL-4; the suspension cells were induced to differentiate into CIK cells with IL-1, IL-2, IFN-γ and CD3 mAb. KG-1a cells were frozen-thawed and the lysate antigen was obtained; the DCs pulsed with or without lysate antigen were co-cultured with CIK (Ag-DC-CIK, DC-CIK groups), and the ratios of CD34+CD38-CD123+LSCs were measured by flow cytometry in different groups; and CIK cultured alone served as blank control. DC-CIK or Ag-DC-CIK was further co-cultured with KG-1a cells, and the apoptotic rates of KG-1a and CD34+CD38-CD123+ cells were also examined by flow cytometry in different groups. Results: DCs were successfully induced from the peripheral blood mononuclear cells. The ratios of CD3+CD56+ cells in CIK, DC-CIK and Ag-DC-CIK groups were sequentially increased to (17.36±4.44)%, (28.22±3.66)%, and (36.16±5.88)%, respectively (P<001). Compared with the control group, the ratio of CD34+CD38-CD123+ in DC-CIK and Ag-DC-CIK groups were significantly decreased (\[3.95±0.53\]%, \[3.03±0.62\]% vs \[8.78±0.62\]%, P<0.01). DC-CIK induced apoptosis of KG-1a cells, with the apoptotic rate increased from (2.34±0.74)% to (12.27±1.01)%, but it showed no evident effect on apoptosis of CD34+CD38-CD123+ cells. Conclusion: DCs co-cultured with CIK can effectively kill acute myelogenous leukemic stem cells, but have no evident proapoptosis effect.

甘肃省科技支撑计划资助项目(No.0804NKCA115)