CAJ | 학술논문

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 killer cells,CIK)和树突状细胞(dendritic cells,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14 d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.
목적 연구래원우간암환자외주혈경세포인자유도적살상세포(cytokine-induced killer cells,CIK)화수돌상세포(dendritic cells,DC)공배양유도적DCIK세포체외항간암세포활성.방법 채집23례간암환자외주혈단개핵세포(peripheral blood mononuclear cell,PBMC),상규유도출DC、CIK.부분CIK여DC안일정비례공배양,획득DCIK세포.배양14 d후,류식세포의검측CIK、DCIK세포표형,MTT법검측CIK화DCIK대인간암SMCC-7721화HepG2세포적살상활성,매련면역법검측CIK화DCIK배양상청중분비적세포인자IL-12、IL-4、IFN-γ.병대각조지간적차이진행통계학분석.결과 DCIK세포고표체CD3+CD8+、CD3+CD56+효응세포,기효응세포적비례、대간암세포적살상활성이급분비적IL-12、IFN-γ적농도교미여DC공배양적CIK세포경고(P＜0.05).결론 여DC공배양가사CIK세포획득경강적체외살간암세포활성,가능여기가분화출경다적효응세포유관.DCIK세포치료가이작위일충림상유효적항간암면역치료책략.
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 peripheral 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-γ in the supernatants of cell culture were detected by enzyme-linked immunosorbent assay (ELISA).Results High expressions of CD3+CD8+ and CD3+CD56+ were observed on DCIKs.The percentages of effector cells,cytotoxic 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 differentiation of effector cells and the cytolytic activities of CIKs against hepatocarcinoma cells in vitro.Immunotherapy with DCIKs may be a promising strategy for the treatment of patients with HCC.