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  

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

作　　者：张丽莉[1] 李世俊[2] 司玉玲[2] 梁芳芳[1] 庞华[2] 王玫 

机构地区：[1]天津医科大学研究生院,300070 [2]天津市第四中心医院肿瘤血液科 [3]中国医学科学院血液病研究所细胞治疗室

出　　处：《天津医药》2012年第5期420-423,共4页Tianjin Medical Journal

基　　金：天津市卫生局科技基金资助项目(项目编号:11KG114)

摘　　要：目的:探讨白血病干细胞(LSC)来源的树突细胞(DC)与细胞因子诱导的杀伤细胞(CIK)共同培养对自身LSC的杀伤作用。方法:提取慢性粒细胞白血病加速期(CML-AP)患者的骨髓单个核细胞(BMMC),再应用CD34+CD38-CD44+标记BMMC,流式分选仪分选出LSC并扩增、诱导出DC。取同源的单个核细胞诱导出CIK。用流式细胞技术检测DC和CIK细胞免疫表型、DC的P-糖蛋白(P-gp)耐药蛋白表达。荧光原位杂交(FISH)技术检测BCR/ABL融合基因在LSC及DC中的表达,乳酸脱氢酶释放法测定效应细胞对靶细胞的杀伤效应。结果:CIK与DC共培养后,CD40、CD80、CD83及人类白细胞抗原(HLA-DR)的构成均高于共培养前;CD3、CD3CD8、CD3CD56的成熟表型表达率高于共培养前,差异有统计学意义(P<0.05或P<0.01)。干细胞来源DC的P-gp表达阳性率为(60.61±12.38)%。3种效靶比对应的杀伤率差异有统计学意义(P<0.01),随着效靶比的增加,细胞杀伤作用增强。结论:白血病干细胞来源DC功能正常,与CIK共培养对自身LSC有杀伤作用。Objective： To investigate the killing effects of stem cell-derived Dendritic Ceils （DC） co-cultured with cytokine induced killer （CIK） on Leukemia Stem Cells （LSC） from the same source. Methods： Bone marrow mononuelear cells were isolated from CML-AP donors, CD34＋CD38-CD44＋ ceils were sorted by flow cytometry, cultured and induced to DC. The mononuelear 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 eytotoxieity activity was measured by lactate dehydrogenase release assay. Results：The expression rates of CD40, CD80, CD83 and HLA-DR were increased in DC co-cultured with CIK; the mature immunophenotypes of CD3, CD3CD8, CD3CD56 were in- creased in CIK co-cultured with DC, the differences were significant （P 〈 0.05 or P 〈 0.01）. The possitive expression rate of DC＇s P-gp was （60.61 ＋ 12.38） %. There were significant differenees of the killing effects in the three groups of effeetor-target ratio （P 〈 0.01）. The killing effect increased with effeetor-target ratio. Conclusion： Leukemia stem cell-derived DC＇s functions were normal, and killing effects can be found when they were co-cuhured with CIK.

关 键 词：树突细胞 白血病 粒-单核细胞 慢性杀伤细胞P糖蛋白 白血病干细胞 

分 类 号：R730.3[医药卫生—肿瘤]