DC-CIK细胞的生物学活性及抗淋巴瘤细胞的作用

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。

The biological activity and anti-tumor effect against lymphoma cells in DC-CIK cells

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) cells 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-γ 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.