DC-CIK细胞的生物学活性及体外抗血液肿瘤研究  

The Study of Biological Activity and Anti-Tumor Effect Against K562 Cells in DC-CIK Cells

作　　者：陈海飞 沈红石 王静 李征洋 唐杰庆 吴天勤 

机构地区：[1]解放军100医院血液肿瘤科南京军区血液肿瘤治疗中心,江苏苏州215006

出　　处：《现代生物医学进展》2012年第30期5823-5826,共4页Progress in Modern Biomedicine

基　　金：全军医药卫生科研项目(09MA016)

摘　　要：目的:研究细胞因子诱导的杀伤细胞(CIK)与树突状细胞(DC)共培养后的体外增殖能力、免疫表型变化、分泌细胞因子水平以及对K562、K562/ADM细胞毒作用的影响。方法:正常人外周血单个核细胞诱导DC和CIK细胞,将DC与CIK共培养,以CIK细胞单独培养为对照。用台盼蓝活细胞计数计算细胞扩增倍数,MTT法测定杀伤活性,流式细胞术分析免疫表型,ELISA双抗体夹心法检测分泌干扰素-γ(IFN-γ)、白细胞介素-12(IL-12)的水平。结果:DC-CIK细胞增殖能力明显高于CIK细胞(P<0.05);DC、CIK细胞共培养后,CD3+CD8+、CD3+CD56+双阳性细胞比率较同条件下CIK细胞组显著增多(P<0.05);共培养3d,DC-CIK细胞上清液中IL-12、INF-7的分泌量均比CIK细胞单独培养的分泌量高(P<0.01,P<0.05);在2.5:1-20:1的效靶比范围内,DC-CIK共培养物对K562和K562/ADM的杀伤活性均高于单纯CIK细胞组,且差异显著(P<0.05),且杀伤率与效靶比呈正相关。结论:DC-CIK细胞的增殖能力、分泌细胞因子水平、对K562和K562/ADM的杀伤活性均高于CIK细胞,为DC-CIK细胞免疫治疗提供了实验和理论依据。Objective： To investigate the proliferation activities,phenotype changes,level of secretory cytokines and anti-tumor activity against K562 cells in co-culture of cytokine-induced killer （CIK）cells with dendritic cells （DC）. Methods： DC and C1K were prepared from healthy human peripheral blood mononuclear cells. They were co-cultured peripherally. CIK cells were cultured alone as controls. Increased number of cells were counted by tapan-blue staining, killing activity was detected by MTT assay, cells phenotypes were analyzed by flow cytometry, secretions of 1NF-~/,and IL-12 were determined by ELISA. Results： The proliferation activity of DC-CIK cells was significantly higher than that of CIK cells （P〈 0.05）. Under the same condition, The ratio of CD3＋ CD56＋ and CD3＋ CD8＋ double positive cells in CIK cells was significantly enhanced by co-culture with DC （P〈 0.05）. In DC-CIK cells, the level oflL-12 and INF-~/, in culture supernatants was increased noticeably on flay 3 compared to CIK cells which were cultured alone（l＇〈0.01 3〈0.05）. At the effector-target ratio from 2.5： 1 to 20：1, the anti-tumor effect of DC-CIK cells was much higher than that of CIK cells（P〈 0.05）, and this effect was positively related with the effector-target ratio. Conclusion： The proliferation activity, level of secretory cytokines, and anti-tumor effect against K562 and K562/ADM ceils of DC-CIK cells were significantly higher than those of CIK cells.The research provides theoretical and experimental basis for the immunotherapy of DC-CIK cells.

关 键 词：树突细胞 杀伤细胞 K562 K562 ADM细胞 

分 类 号：Q813[生物学—生物工程] R730.231[医药卫生—肿瘤]