摘要
目的研究来源于肝癌患者外周血经细胞因子诱导的杀伤细胞（cytokine—induced kil—lercells，CIK）和树突状细胞（dendritic ceils，DC）共培养诱导的DCIK细胞体外抗肝癌细胞活性。方法采集23例肝癌患者外周血单个核细胞（peripheral blood mononuclear cell，PBMC），常规诱导出DC、CIK。部分CIK与DC按一定比例共培养，获得DCIK细胞。培养14d后，流式细胞仪检测CIK、DCIK细胞表型，MTT法检测CIK和DCIK对人肝癌SMCC-7721和HepG2细胞的杀伤活性，酶联免疫法检测CIK和DCIK培养上清中分泌的细胞因子IL-12、IL-4、IFN-γ。并对各组之间的差异进行统计学分析。结果DCIK细胞高表达CD3^＋CD8^＋、CD3^＋CD56^＋效应细胞，其效应细胞的比例、对肝癌细胞的杀伤活性以及分泌的IL-12、IFN-γ的浓度较未与DC共培养的CIK细胞更高（P〈0．05）。结论与DC共培养可使CIK细胞获得更强的体外杀肝癌细胞活性，可能与其可分化出更多的效应细胞有关。DCIK细胞治疗可以作为一种临床有效的抗肝癌免疫治疗策略。
Objective To explore in vitro cytotoxic activities of DCIKs against hepatocarcinoma cells by co-culturing cytokine-induced killer cells （CIKs） with dendritic cells （DCs） derived from peripher- al blood of patients with hepatocellular carcinoma （HCC）. Methods Peripheral blood mononuclear cells （PBMCs） were isolated from 23 patients with HCC and cultured with cytokines to induce DCs and CIKs. DCIKs were induced by co-culturing CIKs with DCs. After 14 days of co-culture, the phenotypes of DCIKs and CIKs were analyzed by flow cytometry, and their in vitro cytotoxic activities against SMCC-7721 and HepG2 hepatocarcinoma cells were measured by MTT assay. Levels of IL-12, IL-4 and IFN-γin the super- natants of cell culture were detected by enzyme-linked immunosorbent assay （ELISA）. Results High ex- pressions of CD3 ^＋ CD8^＋ and CD3 ^＋ CD56^＋ were observed on DCIKs. The percentages of effector cells, cytotox- ic activity and cytokine secretion were all significantly increased with DCIKs as compared with those CIKs without DC co-culture （P〈0.05）. Conclusion Co-culture of CIKs with DCs can enhance the differentia- tion of effector cells and the cytolytic activities of CIKs against hepatocarcinoma cells in vitro. Immunothera- py with DCIKs may be a promising strategy for the treatment of patients with HCC.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2014年第1期42-46,共5页
Chinese Journal of Microbiology and Immunology
