CIK与树突细胞联合对肾癌细胞的杀伤研究

目的研究细胞因子诱导的杀伤细胞（CIK）和树突状细胞（Dc）共同培养后CIK细胞的增殖活性、表型变化及对肾癌细胞株769一P的杀伤作用。方法采集健康人的外周血单个核细胞（PBMC），置37℃、5％C02培养箱2h，收集非粘附细胞诱导分化成CIK，粘附细胞诱导为DC，在培养7d后DC与CIK按1：40的比例混合培养，分别在3d、6d收集细胞同时以单纯CIK细胞为对照，运用LDH释放法检测对769一P细胞的杀伤活性。结果Dc—CIK共同培养后增殖速度明显快于单纯的CIK细胞，培养第10天时单纯CIK的CD3CD8、CD；cD未双阳性率分别为（50．4±1．8）％、（20．1±5．2）％，共培养3dDC—CIK的CD；CD8＋、CD36＋未的阳性率分别为（67．2±5．2）％、（37．9±4．1）％，6dDC-CIK的CD3＋CD8、CD3CD56的阳性率分别为（75．2±3．1）％、（48．3±2．9）％，表达差异具有统计学意义（P〈0．05）。在1：5—1：40靶效比范围内DC—CIK对肾癌细胞的杀伤活性高于单纯CIK细胞，且随效靶比增强杀伤活性增强（P〈0．05）。结论DC—CIK的增殖活性和细胞毒活性均高于单纯CIK细胞。

Cytotoxicity Effect of Co-Cultured Dendritic Cells with Cytokine Induced Killer Cells on Renal Cancer Cells in Vitro

Objective To study of the cytotoxicity effect of co-cultured dendritic cells with cytokine induced killer cells on hu- man renal cancer cells of 769-P. Methods Peripheral blood mononuclear cells （PBMC） were isolated from healthy adult donors, and based on the adhesion characteristics of the cells, DCs were induced from adherent cells by some certain cytokines ,while non- adherent cells were induced to CIK by some another cytokines. After 7 days incubation DCs were co-cultured with CIKs at a ratio of 1 to 40 for 3 days,whose cytotoxicity activity on cells of 769-P was detected by lactate dehydrogenase（LDH） assay while com- paring with simple CIKs. Results Compared with the simple CIKs,the co-cultured DC-CIKs presented a markedly higher prolif- eration. On 10th of incubation, the positive rates of CD3 CD8＋ ,CD3 CD56＋ cells were （50.4 ±1.8） %, （20.1± 5.2） % in CIKs. On 3rd of incubation the positive rates of （ 67.2 ±5.2 ） % , （37.9 ±4. 1 ） % on 3rd of incubation and on 6th of incubation （75.2 ± 3.1 ） % , （48.3 ± 2.9 ） % in co-euhured DC-CIKs, showing a significant differences between two groups （ P 〈 0.05 ）. Within the target range of 5： 1 to 40： 1 the cytotoxicity effect of DC-CIKs on renal cancer cells was higher than CIKs＋ （P 〈 0.05）. Conclusion The proliferation and Cytotoxicity of co-cultured DC-CIKs are both higher than CIKs.