体外培养DC-CIK细胞杀伤白血病细胞实验研究-中国肿瘤生物治疗杂志.docVIP

体外培养DC-CIK细胞杀伤白血病细胞实验研究 张素芳，郭晓玲，张娜娜，张静楠，任金海，郭晓楠 （河北医科大学第二附属医院 血液科， 河北 石家庄050000） 摘 要:目的:未缓解的急性白血病患者的外周血中存在一定比例的白血病细胞，能否诱导CIK细胞的增殖用于复发难治白血病的挽救治疗值得探讨。体外诱导培养缓解期及未缓解期急性白血病患者来源的DC-CIK细胞，比较其增殖能力，免疫表型及杀瘤活性的变化。方法:提取复发难治白血病患者和缓解期患者的外周血单个核细胞，按常规方法诱导产生DC-CIK细胞，在培养过程中计算细胞增殖倍数，用流式细胞仪检测CD3CD8、CD3CD56双阳性率，并在细胞增殖期检测比较其对K562细胞及患者单个核细胞的杀伤活性。结果:缓解组和未缓解组来源的DC-CIK细胞均不断快速增殖，培养第15天时，缓解组CD3+CD8+和CD3+CD56+细胞分别占(87.5±1.29）%，(63.8±1.98）%，未缓解组分别占(88.6±3.4)%,(66.8±4.1)%，此时应用CCK-8法测得在效靶比为20:1时，缓解组对K562细胞的杀伤率为（42.64±3.25）%，未缓解组为（57.49±2.87）%；相同效靶比下，未缓解组对患者的单个核细胞的杀伤率为（67.29±7.95）%。结论:应用未缓解患者的外周血单个核细胞,同样可以培养出DC-CIK细胞，具有与缓解组来源的DC-CIK细胞一致的高增殖活性和高CD3+CD8+、CD3+CD56+表达，一样对K562细胞杀伤活性较强，并对患者单个核细胞具有特异杀伤活性，未缓解患者来源的CIK细胞中未检测到白血病细胞残留，可以应用于难治白血病的挽救治疗。 关键词:细胞因子诱导的杀伤细胞；树突状细胞；生物学活性；白血病；细胞因子 Biological Activity of DC-CIK Cells And effect against Leukemia Cells In Vitro ZHANG Su-fang，GUO Xiao-ling，ZHANG Na-na，ZHANG Jing-nan，REN-Jinhai，GUO Xiao-nan （Hebei Medical University The Second Affiliated Hospital,Shijiazhuang of Hebei，050000，China） DC-CIK cells source from remission and non-remission of acute leukemia patients Induced in vitro ,compared their changes of proliferation,immune phenotype and tumoricidal activity .Methods：Peripheral blood mononuclear cells of objects were extracted and induced by conventional methods into DC-CIK cells,calculated proliferation multiples,CD3+CD8+,CD3+CD56+ double-positive rate detected by flow cytometry,and killing activity against K562 cells and mononuclear cells was compared in cell proliferation.Results：DC-CIK cells come from Remission and non-remission group were continued proliferating rapidly,on the day of 15 cultured ,remission group cells with CD3+CD8+ and CD3+CD56+ accounted for(87.5±1.29)%, (66.8±4.1)%,this time measured by CCK-8 in the target ratio of 20:1, the killing activity of remission group to K562 was (42.64±3.25)%,non-remission group (57.49±2.87)%;the same effector to target ratios,non-remission group to mononuclear cell of patients was(67.29