【摘要】 目的研究脐血树突状细胞-细胞因子诱导的杀伤(DC-CIK)细胞的体外增殖、免疫表型、分泌细胞因子水平及其对急性白血病细胞细胞毒作用的影响。方法采集脐血单个核细胞诱导DC和CIK细胞。将DC和CIK细胞按1∶5的比例混合培养,以脐血CIK细胞或外周血DC-CIK细胞为对照。用流式细胞术分析细胞表型,台盼蓝活细胞计数计算细胞扩增倍数,MTT法检测效应细胞杀伤白血病细胞的活性,ELISA法测定分泌干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-12(IL-12)的水平。结果脐血DC-CIK细胞的增殖能力显著高于脐血CIK细胞和外周血DC-CIK细胞(P均<0.05);脐血DC、CIK细胞共培养后,CD3+CD8+、CD3+CD56+细胞比例较同条件下CIK细胞明显增多(P<0.05);混合培养3 d,脐血DC-CIK细胞上清液中IL-12、IFN-γ、TNF-α含量均比CIK细胞单纯培养的分泌量高(P<0.01或P<0.05);在2.5∶1～20∶1的效靶范围内,脐血DC-CIK细胞对各亚型急性白血病细胞的杀伤率明显高于CIK细胞(P<0.05),且对各亚型白血病细胞杀伤活性无统计学意义,与外周血DC-CIK细胞对白血病杀伤效应相类同。结论脐血DC可增强同源CIK细胞的增殖活性和抗白血病效应。脐血DC-CIK细胞增殖能力比外周血DC-CIK细胞强,但两者在细胞毒方面无显著性差异。脐血来源丰富,且输注不易引起严重的移植物排斥反应,其DC-CIK细胞在免疫治疗方面应有更广泛的临床应用前景。更多还原

【Abstract】 Objective To study the proliferation capability in vitro,immunophenotype changes and secreted cytokines level of cord blood DC-CIK cells and their effect on cytotoxicity of acute leukemia cells.Methods DCs and CIK cells were induced from cord blood mononuclear cells.They were co-cultured according to the ratio of 1∶5,and CIK cells from cord blood or DC-CIK cells from peripheral blood were cultured as controls.Immunophenotype changes were analyzed by flow cytometry,the increased number of cells was counted by trypan-blue staining,and the killing effect on leukemia cells was detected by MTT assay.The interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α) and interleukin-12(IL-12) levels of the cultured supernatants were detected by ELISA kits.Results The proliferation capability of cord blood DC-CIK cells was significantly higher than that of cord blood CIK cells and peripheral blood DC-CIK cells(both P<0.05).Under the same condition,the ratio of double positive cells CD3+CD8+ and CD3+CD56+ in CIK cells was significantly enhanced when co-cultured with cord blood DC cells(P<0.05).The levels of IL-12,IFN-γ and TNF-α in cultured supernatants of cord blood DC-CIK cells increased noticeably on day 3 as compared with those of CIK cells cultured alone(P<0.01,P<0.05,P<0.05).Within the effector-target ratio range from 2.5∶1 to 20∶1,the killing capacity of cord blood DC-CIK cells against all subtypes of acute leukemia cells was much higher than that of CIK cells(P<0.05),but there was no significant difference among all subtypes of acute leukemia cells.It was the same as the killing effect of peripheral blood DC-CIKcells on leukemia cells.Conclusion The proliferation capability and anti-leukemia effect of the homologous CIK cells can be enhanced by cord blood DCs.The proliferation capability of cord blood DC-CIK cells is stronger than that of peripheral blood DC-CIK cells,but there is no significant difference in cytotoxicity between them.Because cord blood can be easily obtained and is not likely to cause serious graft rejection,DC-CIK cells have more extensive clinical application in immune therapy.更多还原