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DC-CIK细胞的体外增殖能力、免疫表型变化、分泌细胞因子水平以及抗淋巴瘤细胞活性。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3 d,DC-CIK细胞上清液中IL-12、INF-γ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5∶1-40∶1的效靶比范围内,DC-CIK细胞对淋巴瘤细胞的杀伤率显著高于CIK细胞(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、抗淋巴瘤细胞活性均高于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 lymphoma cells of DC-CIK cells in co-culture of cytokine-induced killer （CIK） ceils with dendritic cells （DC）. Methods DC and CIK 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 INF-γ 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 of IL-12 and INF-7 in culture supernatants was increased noticeably on day 3 compared to CIK cells which were cultured alone （P〈0.01, P〈0.05）. At the effector-target ratio from 5 ： 1 to 40 = 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 lymphoma cells 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.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2007年第3期283-285,318,共4页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省"三五人才工程"专项基金资助项目
关键词
CIK细胞
DC-CIK细胞
生物学活性
抗淋巴瘤
cytokine-induced killer cell
dendritic cells-cytokine induced killer cell
biological activity
anti-lym- phoma
