摘要
目的 通过恶性胸腔积液获取成熟树突状细胞（DC）,探讨恶性胸腔积液来源的DC与细胞因子诱导杀伤细胞（CIK）共培养对CIK细胞的作用及两者共培养后联合奥沙利铂（L-OHP）对肺腺癌的体外杀伤作用.方法 收集20例非小细胞肺癌（NSCLC）患者的胸腔积液,每例收集800～1 000 ml,双层聚蔗糖（Ficoll）密度梯度离心获得DC前体细胞,体外诱导培养收获成熟DC;分离健康人外周血单个核细胞（PBMC）,体外培养获得CIK细胞;DC与CIK细胞共培养;流式细胞仪鉴定表型;MTT法检测对肺腺癌的体外杀伤作用.结果 ①恶性胸腔积液中存在DC前体细胞,经体外细胞因子诱导培养后可获得成熟的DC,培养9天的DC表面标志物CD80、CD83、CD86及MHC-Ⅱ类分子HLA-DR与第0天比较明显增高,分别为（56.61±7.01） % vs （14.38±5.16）%,（58.85±7.05）% vs （18.49±9.43）%、（60.27±13.94）% vs （20.39±6.67）%、（70.97±8.96）% vs （41.36±8.57）%（均P＜0.01）.②培养14天后,DC与CIK细胞共培养较单独CIK培养中CD3＋/CD56＋、CD3＋、CD4＋、CD8＋细胞明显增多（均P＜0.01）;对肺腺癌细胞杀伤作用明显增强（P＜0.01）.③DC与CIK细胞共培养联合L-OHP治疗对肺腺癌细胞的杀伤率明显高于单独治疗（P＜0.01）.结论 胸腔积液来源的DC前体细胞在体外诱导培养可获得功能正常的成熟DC,与CIK细胞共培养后促进CIK细胞增殖、增强其杀伤作用;两者共培养后联合L-OHP具有协同作用,对肺腺癌在体外有明显的抑制作用,为肺癌综合治疗提供了一定的理论基础.
Objective Getting mature dendritic cells（DCs） by malignant pleural effusion,to establish the effect of malignant pleural effusion derived dendritic cells co-cultured with eytokine-indueed killer （CIK） combined with oxaliplatin（L-OHP） for lung cancer cell. Methods Precursory DCs were obtained by adherence after density gradient centrifugation of malignant pleural effusion of 20 patients with non-small cell lung cancer, and were cultured with cytokines. CIK cells were prepared routinely from human peripheral blood mononuelear cells（PBMC）. Flow cytometry was used to explore the immunophenotypes,and in vitro cytotoxieity was examined by MTT assay. Results （DDCs of malignant pleural effusion had the typical morphology after extracorporeal cultivation, cultivate nine days of DCs surface markers CD80,CD83,CD86 and MHC-II class molecules HLA-DR showed higher expression compared with 0 days, respectively,（56.61±7.01） % vs （14.38±5.16）%,（58.85±7.05）% vs （18.49±9.43）%,（60.27±13.94）% vs （20.39±6.67） %,（70.97±8.96）% vs （41.36±8.57）%（all P 〈0.01）. （2）After co-culture with DC 14 days,the cell surface marker CD3±/CD56±, CD3±, CD4±, CD8± were significantly increased than the CIK cultured alone（ P〈 0.01）. （3）DC-CIK ceils combined with LOHP obviously inhibited the growth of lung adenoeareinoma cells in vitro（ P〈 0.01）. Conclusion Pleural effusion source of DC precursor cells cultured in vitro maturation availability functioning DC,after CIK cells co-cultured with CIK promote cell proliferation, enhance its killing effect; two co-cultured with L- OHP combined synergistic effect, lung cancer was significantly inhibited in vitro, comprehensive treatment of lung cancer provides theoretical foundation.
出处
《临床荟萃》
CAS
2014年第4期385-388,共4页
Clinical Focus
基金
国家自然科学基金资助项目(30800514)
关键词
肺肿瘤
胸腔积液
恶性
树突细胞
细胞因子诱导杀伤细胞
lung neoplasms
pleural effusion, malignant
dendritic cells
cytokine-induced killer cells
