人肺癌细胞NHE—1基因片段的克隆及其反义表达载体的构建

目的:动态观察CIK(cytokine induced killer)细胞的体外增殖,体外的细胞毒活性,及通过动物实验研究其体内的抗肿瘤作用.方法:通过提取健康供血者的PBMC,第0天加入γ-IFN,第1天加入IL-2、抗-CD3单抗和IL-1培养CIK细胞;在流式细胞仪上做动态培养物的表型分析;与LAK细胞作对比,分别用MIT法测定其体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用.结果:CIK细胞在培养2周后获得大量增殖,表型分析表明,CIK细胞属异质性细胞群,在培养的过程中,群体的CD3~ CD56~ 细胞大量扩增达1000多倍,是CIK细胞的主要效应细胞;实验证明,CIK细胞的体外细胞毒活性及对S180荷瘤鼠的体内抗肿瘤作用均强于LAK细胞;其较强的体内抗癌活性可能与荷瘤鼠主体内T细胞活化有关.结论:CIK细胞是一种强于LAK细胞的、新型、高效、具有广谱杀瘤活力的免疫活性细胞.

The Proliferation Profile in vitro and Anti-Tumor Effects of CIK Cells in vivo and in vitro

Objective: To investigate the proliferation profile and the cytoloxicity of CIK (cytokine induced killer) cells, in vitro, and their anti-tumor effects on S180 bearing mice in vitro. Methods: CIK cells were generated by culturing PBMC in the presence of r-IFN on day 0 and IL-2, anti-CD3 McAb, IL-1 on day 1; CIK cultures were analyzed on different time points by FACS; Compared with LAK cells, the cytotoxicity in vitro by MTT assays and the anti-tumor effects on S180 bearing mice in vivo of CIK cells were tested respectively. Results: The flow cytometry analysis showed that CD3 CD56 T cells which were the major cytolytic effectors in CIK cells expanded 1 000-fold after 2 weeks in CIK culture ; CIK cells had greater cytotoxicity against tumor cells in vitro than that of LAK cells; and had also significantly anti-tumor effects in mice bearing S180 tumor compared with LAK cells in vivo. (median inhibitory rates 78% vs 33%, P < 0.05 t ); The CIK therapy was possibly correlated with induction of the activation of the splenic T lymphocytes of S180 bearing mice. Conclusion: CIK cells were highly efficient cytolytic effector cells which were non-MHC restricted.