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来源于干细胞的树突细胞激活免疫细胞对同源白血病干细胞的杀伤作用

张丽莉1,2,李世俊1,司玉玲1,梁芳芳1,庞华1   

  1. 1. 天津市第四中心医院
    2. 天津医科大学
  • 收稿日期:2011-11-21 修回日期:2012-03-03 出版日期:2012-05-15 发布日期:2012-05-15
  • 通讯作者: 庞华

The killing effects of immune cells induced by Leukemia Stem Cell-derived Dendritic Cells on Leukemia Stem Cells from the same source

  • Received:2011-11-21 Revised:2012-03-03 Published:2012-05-15 Online:2012-05-15

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摘要： 摘要 目的：探讨白血病干细胞（LSC）来源的树突状细胞（DC）与细胞因子诱导的杀伤细胞（CIK）共同培养对自身白血病干细胞的杀伤作用。方法：提取慢性粒细胞白血病加速期（CML-AP）患者骨髓单个核细胞，应用CD34+CD38-CD44+标志，流式分选白血病干细胞，并扩增、诱导出DC。取同源的单个核细胞诱导出CIK。DC与CIK共同培养后的细胞为效应细胞，白血病干细胞为靶细胞，检测在不同效靶比时效应细胞对靶细胞的杀伤作用。用流式细胞技术检测DC和CIK细胞免疫表型、DC的P-gp耐药蛋白表达。FISH方法检测BCR/ABL融合基因在白血病干细胞及DC中的表达，乳酸脱氢酶释放法（LDH释放法）测定杀伤效应。结果：DC和CIK细胞培养成熟并能表达成熟DC、CIK免疫表型。DC的P-gp耐药蛋白表达阳性，BCR/ABL融合基因阳性。杀伤效应随着效靶比的增高而增强。效靶比40:1时杀伤效应最强，在此效靶比下，效应细胞对靶细胞的杀伤率为（59.13±12.82）%。结论：白血病干细胞来源DC功能正常，与CIK共培养对自身白血病干细胞有杀伤作用。

关键词: 树突状细胞, 慢性粒细胞白血病, 白血病干细胞, 细胞因子诱导的杀伤细胞, P-糖蛋白

Abstract: Abstract Objective: To investigate the killing effects of stem cell-derived Dendritic Cells (DC) co-cultured with cytokine induced killer (CIK) on Leukemia Stem Cells（LSC）from the same source. Methods: Bone marrow mononuclear cells were isolated from CML-AP donors, CD34+CD38-CD44+ cells were sorted by flow cytometry, cultured and induced to DC. The mononuclear cells from the same doner were induced to CIK. DC and CIK were co-cultured to determined killing effects. The expressions of DC's and CIK's markers typical、P-gp resistance protein were detected by flow cytometry. The expressions of bcr/abl fusion gene of leukemia stem cells and DC were detected by fluorescence in situ hybridization (FISH). The cytotoxicity activity was measured by lactate dehydrogenase release assay. Results: DC and CIK showed typical immunophenotype. DC expressed P-gp resistance protein, and DC's bcr/abl fusion gene was positive. The killing effect increased with effector-target ratio. It was showed that under an effector-target ratio of 40:1 the killing effect was highest. At this level, killing rate can reach to (59.13±12.82)%. Conclusion: Leukemia stem cell-derived DC's functions were normal, and killing effects can be found when they were co-cultured with CIK.

Key words: dendritic cells, chronic myelogenous leukemia, Leukemia Stem Cells, cytokine induced killer, P-glycoprotein