树突状细胞联合细胞因子诱导的杀伤细胞体外抗肺癌的实验研究  

The Anti-lung cancer effects of dendritic cells associated with cytokine-induced killers in vitro and in vivo

作　　者：周菁[1] 张伟[1] 

机构地区：[1]南昌大学一附院呼吸内科,南昌330006

出　　处：《江西医药》2009年第11期1064-1066,共3页Jiangxi Medical Journal

摘　　要：目的探讨树突状细胞(DC)联合细胞因子诱导的杀伤细胞(CIK)在体内外能否有效的抗肺癌。方法正常人外周血单个核细胞经不同细胞因子作用后培养成DC和CIK细胞,单个核细胞经贴附塑料平皿获得单核细胞,加入GM-CSF,IL-4及TNF-a诱导获得DC,悬浮细胞加入IFN-g,24h后IL-1a,IL-2及CD3McAb诱导获得CIK,将A549制备成肿瘤细胞冻融物,作为肿瘤抗原刺激DC,并将DC与CIK细胞联合培养(DC+CIK,负载抗原的DC+CIK),以CIK细胞单独培养作为对照。Cytotoxi-city TOX96体外杀伤实验测定体外细胞毒活性。结果在培养的第15d,与负载抗原的DC共同培养的CIK与CIK细胞单独培养细胞相比,增殖速率明显提高[(23.4±2.3)倍vs(16.7±2.7)倍,P<0.05],CD3+CD56+细胞表达水平也明显提高,[(64.3±3.6)%vs(43.9±2.1)%,P<0.05],同时负载抗原的DC的CIK对A549细胞的体外下细胞毒活性增强。结论DC与CIK细胞共培养后可提高CIK细胞的增殖速率,提高CIK细胞表型的表达水平,增强CIK抗肺癌的活性,将来可作为一种临床有效的过继免疫治疗策略。Objective To explore whether dendritic cells（DC） associated with cytokine-induced killers（CIK） resist against lung cancer cells effectively in vitro and in vivo.Methods DC and CIK were prepared from human peripheral blood mononuelear cells （PBNC） with different cytokines. The tumor cell line A549 prepared into carcinoma cell lysates were burdened on DCs as tumor antigens.The DCs pulsed with or without A549 cell lysates were eocuhure with CIK.CIK was used as control. Cells phenotypes were analysed by flow cytometry （FCM）.T-cell proliferating activity stimulated by DCs was identified by auto-mixed lymphocyte reaction and -mixed lymphocyte reaction（MLR）.Cytotoxicity in vitro was assayed by Cytotoxicity TOX96 assay.Results Compared with CIK ,L-DC-CIK got a significant increasing of proliferation rate[ （23.4±2.3）times vs （ 16.7±2.7 ）times , P〈0.05] and ,CD3＋CD56＋ expression rate [（64.3±3.6）%vs （43.9±2.1）%,P〈0.05].Meanwhile L-DC-CIK led to an increase of cytotoxie activity to A549 cells and showed significang inhibition of growth of transplanted tumor cells.Conclusion Coculture of CIK with DC can promote the proliferation effectivity ,cell phenotypes and cytotoxicity of CIK against tumor in vitro .DC-CIK is promising as adoptive immuno-therapeutic strategy for patients of lung cancer.

关 键 词：树突状细胞 细胞因子诱导的杀伤细胞 A549细胞系 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]