关键词: 树突状细胞;LAK细胞;免疫治疗

    摘 要:目的 建立体外培养扩增树突状细胞(DC)的方法，并观察其诱导的抗肿瘤免疫作用. 方法 分离外周血单个核细胞，应用GM-CSF及IL-4诱导DC产生，用TNF-α和PGE2 促进其成熟，并用肺癌细胞系GLC-82可溶性抗原致敏.将成熟DC与自体LAK细胞混合培养(DC-LAK)，用MTT法检测DC-LAK细胞及LAK细胞对多种肿瘤细胞系的杀伤作用. 结果 经GM-CSF，IL-4，TNF-α，PGE2 作用后，细胞表达CD1 a阳性率为71.0%，CD83 50.0%，CD14 阳性率低于10.0%，高表达HLA-Ⅰ，HLA-Ⅱ和CD45 ，为典型的DC表型特征.DC-LAK和LAK细胞对4种肿瘤细胞(GLC-82，A549，moser和MCF-7)的杀伤率，前者为84.7%，66.9%，49.3%和56.0%，后者为43.5%，31.3%，45.0%和32.4%. 结论 成功的诱导出具有典型形态特征及增强LAK细胞杀伤活性的DC.

    In vitro generation of human blood dendritic cells for active tumor immunotherapy

    ZHANG Jian-Fang，YU Wen-Bin，XU Xiu-Li，SU Ming-Quan，MA Yue-Yun，LIU Jia-Yun，Ding Zhen-Ruo

    Center of Clinical Molecular Biology，Xijing Hospi-tal，Fourth Military Medical University，Xi'an710033，China

    Keywords:dendrtic cells;LAK;immunotherapy

    Abstract:AIM To establish a method to culture dendritic cells(DC)in vitro and to observe the anti-tumor effect in-duced by DC.METHODS Dendritic cells were derived from PBMC stimulated with GM-CSF and IL-4and DC were pulsed with tumor soluble antigen abstracted from lung can-cer cell line GLC-82.TNF-αand PGE2 were used to enhance antigen presenting ability of DC.Phenotype of DC were ana-lyzed through FACS.Mature DC were cocultured with LAK cells and the mixed cells were named DC-LAK.Cytotoxicity of DC-LAK and LAK were measured by MTT assay.RESULTS The cultured DC expressed HLA-Ⅰ ......