共培养的树突细胞和CIK细胞对肺癌的体内外抑癌作用

背景与目的:细胞因子诱导的杀伤(cytokine-inducedkiller,CIK)细胞是高效的肿瘤杀伤细胞。树突细胞(dendriticcells,DCs)是体内最强的抗原递呈细胞,并且能够提高效应细胞的抗瘤活性。本实验将DCs和CIK细胞共培养观察DCs对CIK细胞的细胞表型、增殖活性及体内外的抗肺癌作用的影响。方法:从健康人外周血单个核细胞中常规诱导出DCs、CIK细胞后,将DCs和CIK细胞按1∶10比例共培养5天获得DC-CIK细胞。流式细胞仪测DC-CIK细胞表型变化,3H-TdR掺入法测定其体外的细胞毒活性,并用肺腺癌细胞株A549建立裸鼠模型观察DC-CIK体内的抗肿瘤效果。结果:在培养第14天,DC-CIK细胞与单独CIK细胞培养组相比,增殖速率提高(17.0±1.8)倍vs.(10.9±2.0)倍,P<0.05],CD3 CD56 表达水平明显上调(36.0±4.2)%vs.(25.7±2.9)%,P<0.05],同时对A549细胞的细胞毒活性明显增强(P<0.05)。裸鼠体内实验表明,接种肺癌细胞51天后DC-CIK组、CIK组的抑瘤率分别为62.9%、41.5%,与对照组相比DC-CIK组及CIK组均抑制裸鼠皮下移植瘤的生长(P<0.01),且DC-CIK组与CIK组抑瘤效应差异有统计学意义(P<0.05)。结论:DCs与CIK细胞共培养可使CIK细胞获得更高的增殖活性和更强的抑癌作用。

Antitumor effects of cocultured dendritic cells and cytokine-induced killer cells on lung cancer in vitro and in vivo

BACKGROUND & OBJECTIVE: Cytokine-induced killer (CIK) cells have high cytotoxic activity against tumor cells. Dendritic cells (DCs) are the strongest antigen-presenting cells (APC) and could increase the cytotoxic activity of immunologic effector cells against tumor cells. This study was to investigate the changes of phenotype, proliferative activity, and in vitro and in vivo cytotoxicity of CIK cells after in vitro co-culturing with DCs. METHODS: DCs and CIK cells were prepared routinely from human peripheral blood mononuclear cells (PBMC), then CIK cells were cocultured with autologous DCs for 5 days at a stimulator-to-responder ratio of 1:10 to prepare immunologic effector DC-CIK cells. Phenotypes of DC-CIK cells were analyzed by flow cytometry; The in vitro cytotoxicity of DC-CIK cells was detected by 3H-TdR incorporation method; the in vivo antitumor activity was evaluated in BALB/c nude mice bearing A549 lung cancer. RESULTS: At the 14th day of culture, compared with CIK cells, DC-CIK cells got a significant increase of proliferation rate (17.0+/-1.8) times vs. (10.9+/-2.0) times, P<0.05] and CD3+CD56+ expression rate (36.0+/-4.2)% vs. (25.7+/-2.9)%, P<0.05], and resulted in an enhancement of cytotoxicity to A549 cells (P<0.05) in vivo. At the 51st day after inoculation of tumor cells, the inhibition rate was significantly higher in DC-CIK group than in CIK group (62.9% vs. 41.5%, P<0.05). DC-CIK cells and CIK cells showed significant inhibitory effects on the growth of transplanted tumor cells as compared with control group (P<0.01). CONCLUSION: DC-CIK cells have higher proliferative activity and cytotoxicity in vitro and in vivo against lung cancer in comparison with CIK cells alone.