树突细胞对免疫活性细胞抑癌效应非限制性增强作用.doc

树突细胞对免疫活性细胞抑癌效应的非限制性增强作用 刘宝瑞 钱晓萍 邹征云 王立峰 葛少华 南京大学医学院附属鼓楼医院肿瘤科，江苏 南京 210008 [摘要] 目的 研究抗原负载的树突细胞对免疫活性细胞抑癌的增强作用。方法 在建立树突细胞体外扩增的基础上，将抗原负载的树突细胞作用于单个核细胞及高聚金葡素活化的单个核细胞，观察其对人肺癌GLC-82和A549细胞的体外和裸鼠体内抑癌效果。结果 抗原负载的树突细胞能显著增强单个核细胞及高聚金葡素活化的单个核细胞对GLC-82和A549细胞的抑制作用，该作用不受主要组织相容性复合物基因（major histocompatibility complex, MHC）限制。结论 抗原负载的树突细胞对免疫活性细胞的抑癌作用有MHC非限制性增强效果。 [关键词] 树突细胞/免疫学； 肿瘤； 免疫细胞；免疫活性 [中图分类号] Enhancement of non-MHC restricted anti-tumor effect of immunocytes by dendritic cells LIU Bao-rui, QIAN Xiao-ping, ZHOU Zeng-yun, et al. Department of Oncology ,Drum Tower Hospital, Faculty of medicine, Nanjing University, Nanjing 210008,China Abstract: Objective To study the effect of antigen-pulsed dendritic cells on tumor inhibition induced by immunocytes. Method On the bases of in vitro expansion and identification of dendritic cells, the antigen-imposed dendritic cells were mixed with either mononuclear cells or highly agglutinative staphylococcin activated mononuclear cells, and their anti-tumor effects were studied in vitro and in vivo respectively on the inhibition of human lung cancer cell lines GLC-82 and A549. Result Antigen-pulsed dendritic cells significantly enhanced the inhibition of human lung cancer cell lines GLC-82 and A549 induced by mononuclear cells without MHC restriction. Conclusion Antigen-pulsed dendritic cells play a role in enhance of anti-tumor effect of immunocytes in non-MHC restricted way. Key words: dendritic cells/immunology; neoplasms; immunity,cellular; immunocompetence 树突细胞对免疫活性细胞抑癌效应的非限制性增强作用 树突细胞（dendritic cell, DC）是已知最强的专职抗原提呈细胞，在抗原摄取、加工、提呈以及诱导特异性免疫应答方面发挥着关键性作用。本文作者在成功地建立树突细胞体外扩增技术并用于肿瘤特异性免疫治疗的同时，意外地发现树突细胞对免疫活性细胞的抗肿瘤效应还具有MHC非限制性增强作用。 1．材料与方法 1.1 主要试剂 白细胞介素-4（IL-4）、肿瘤坏死因子（TNF-α）和粒细胞-巨噬细胞集落刺激因子（GM-CGF）购自Promega公司，高聚金葡素（HAS）为沈阳协和集团产品，前列腺环素（PGI2）购自Sigma公司。CD1a-FITC、CD83-FITC为Ancell公司产品，HLA-I、II及羊抗鼠-FITC购自北京帮定公司。 肿瘤可溶性抗原的提取 肺癌GLC-82细胞在含5%小牛血清的RPMI1640培养液中呈对数生长时，收集并离心洗涤细胞，低渗反复冻融4次，5000r/min离心30min，取上清测定蛋白含