肺癌相关抗原RNA转染树突状细胞诱导细胞因子诱导的杀伤细胞杀伤活力的体外研究  

Study of lung cancer associa ted antigen RNA-transfected DC-induced cell killing activity of CIK in vitro

作　　者：刘伯轩[1] 戴北鸿 向启德 廖东承 高细强[1] 吴玉兰[1] 周频[1] 

机构地区：[1]湖南省怀化市第二人民医院,418000

出　　处：《实用医技杂志》2015年第7期695-698,共4页Journal of Practical Medical Techniques

摘　　要：目的研究NCI-H1975人肺腺癌细胞株相关抗原RNA转染的人树突状细胞(DCs)在体外诱导细胞因子诱导的杀伤细胞(CIK)对人肺腺癌细胞的杀伤活力。方法使用血细胞单采机采集外周血单个核细胞(PBMCs)并利用密度梯度离心法纯化获得的单个核细胞,细胞培养瓶贴壁获得单核细胞,添加重组人粒细胞、巨噬细胞集落刺激因子(rh GM-CSF)和重组人白细胞介素-4(rh IL-4)诱导为未成熟DCs,提取NCI-H1975细胞总RNA转染DCs后诱导为成熟DCs。转染诱导后成熟DCs与PBMCs培养的CIK细胞混合培养后诱导,再次与靶细胞混合培养以观察杀伤活力。实验分转染肺癌相关抗原RNA的DC-CIK组、未转染DC-CIK组及转染空脂质体DC-CIK组,分别采用流式细胞分析法检测DC及CIK细胞表面抗原表达、采用CSFE/碘化丙啶(PI)荧光双标法检测3组杀伤肿瘤活性的差异。结果转染相关抗原RNA组、未转染组及转染空脂质体组的DC及CIK细胞表面抗原表达明显上调,转染相关抗原RNA组与其他2组对肺癌原代细胞杀伤活力明显增强。结论肺癌相关抗原RNA转染DC诱导CIK细胞对靶细胞的杀伤活力明显增强,可为临床应用个体化DC-CIK细胞过继免疫治疗提供新的方法。Objective To investigate the NCI-H1975 human lung adenocarcinoma cell line associated antigen RNA-transfected human dendritic cells（DCs）in vitro cytokine-induced killer（CIK）activity induced killer cells to cancer cells. Methods Using blood cell apheresis machine collecting peripheral blood mononuclear cells（PBMCs） and using mononuclear cells by density gradient centrifugation purified,cell culture flasks adherent mononuclear cells,adding recombinant human granulocyte-macrophage colony-stimulating factor（rh GM-CSF） and recombinant human interleukin-4（rh IL-4）to induce immature DCs,extraction of total RNA NCI-H1975 cells were transfected DCs induced to mature DCs.After transfection induce mature DCs and PBMCs cultured CIK cells induced mixed culture,mixed culture with target cells again to observe the destruction of vitality. The experimental points associated antigen RNA-transfected DC-CIK group of lung cancer,non-transfected DC-CIK group and transfected with empty liposomes DC-CIK group were detected by flow cytometry analysis DC and CIK cell surface antigen expression,using CSFE/PI double staining was used to detect differences in anti-tumor activity of the three groups. Results RNA-associated antigen transfected group, DC untransfected group and transfected with empty liposome group and CIK cell surface antigen expression was significantly associated antigen RNA transfection on the group turned in primary cell killing activity and the other two groups significantly increased lung cancer. Conclusion Lung cancer associated antigen RNA-transfected DC induced killer cells to target the vitality of CIK cells was significantly enhanced for the clinical application of individual DC-CIK cells had adoptive immunotherapy offers a new approach.

关 键 词：树突细胞 细胞因子诱导杀伤细胞 免疫疗法 过继 腺癌 细支气管肺泡 

分 类 号：R734.2[医药卫生—肿瘤]