人巨噬细胞炎性蛋白-1β基因修饰肿瘤细胞的体内致瘤性和瘤苗效果  

Effects of hMIP-113 gene modification on in vivo tumorigenicity and vaccine efficacy of tumor cells

作　　者：罗小玲[1] 谢裕安[1] 匡志鹏[1] 吴继宁[1] 梁安民[1] 

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,南宁530021

出　　处：《中华肿瘤杂志》2008年第2期97-102,共6页Chinese Journal of Oncology

基　　金：广西科技厅青年基金资助项目（桂科青0447049）

摘　　要：目的探讨人巨噬细胞炎性蛋白-1β（hMIP-1β）基因修饰小鼠结肠腺癌CT26细胞的致瘤性和瘤苗免疫效果。方法通过重组腺病毒载体介导，将hMIP-1β基因导入CT26细胞中，X-gal染色法检测基因转染效率；采用酶联免疫吸附试验（ELISA）法检测hMIP-1β基因修饰CT26细胞培养上清中hMIP-1β的含量；采用Boyden趋化小室法检测培养上清对CD4^＋细胞、CD8^＋T细胞、NK细胞和未成熟树突状细胞（imDC）的趋化作用。BALB／c小鼠皮下接种hMIP-1β基因修饰的CT26细胞，观察其体内致瘤性的改变和对免疫细胞的趋化作用；制备hMIP-1β基因修饰的CT26细胞瘤苗并免疫BALB／c小鼠，观察其诱导免疫细胞的杀伤活性和保护性免疫反应。结果腺病毒载体可介导hMIP-1β基因转染CT26细胞和表达，X-gal染色的阳性率可达95．0％以上。在培养上清中hMIP-1β水平为980pg／ml细胞，并对CD4^＋T细胞、CD8^＋T细胞、NK细胞和imDC有显著的趋化作用，与转染对照基因LacZ的CT26细胞及野生型CT26细胞比较，差异有统计学意义（P〈0．01）。体内致瘤实验显示，hMIP-1β基因修饰的T26细胞皮下接种后，成瘤率降低，肿瘤生长速度减慢，肿瘤内可见明显坏死灶，坏死灶内和周围可见较多淋巴细胞浸润。hMIP-1β基因修饰的CT26细胞瘤苗免疫小鼠能有效诱导肿瘤特异性CTL活性和非特异性NK活性，产生明显的免疫保护作用，可抵抗肿瘤细胞的再攻击。结论hMIP-1β基因修饰的CT26细胞致瘤性显著下降，其瘤苗可诱导强烈的抗肿瘤免疫反应，提示hMIP-1β基因修饰瘤苗有望成为更有效的抗肿瘤免疫治疗手段。Objective To explore the effects of human macrophage inflammatory protein-1 beta （hMIP-1β） modification on the in vivo tumorigenicity and vaccine efficacy of tumor cells. Methods Murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus carting the hMIP-1β gene （ AdhMIP-1 β）. The efficacy of gene transfection was tested by X-gal staining. The hMIP-1 β level in the supernatant of hMIP-1 β gene-modified CT26 cells was assayed by ELISA, and the chemotactic activity for CD4 T cells, CDs T cells, NK cells and immature dendritic cells （imDCs） were assayed by a transwell chamber. The changes of growth characteristics and in vivo tumorigenicity of hMIP-1β genemodified CT26 cells were also assessed. BALB/c mice were immunized with hMIP-1 β gene-modified CT26 tumor vaccine and the antitumor effect was evaluated. Results hMIP-1 β gene could be transfected into CT26 cells by AdhMIP-1β with an efficiency over 95%. The level of hMIP-1β in the culture supernatant of hMIP-1β gene-modified Cq26 cells was 980 pg/ml and the supernatant displayed ramarkable chemotactic activity to CD4 T cells, CDs T cells, NK cells and imDCs compared with LacZ gene-modified CT26 cells and control. When the hMIP-1 β gene-modifited CT26 cells were subcutaneously inoculated in BALB/c mice, the tumorigencity was delayed and suppressed, and overt necrosis and lymphocyte infiltration were observed in the tumor tissue, but not in those inoculated with LacZ gene-modified C126 cells or parental CT26 cells. The mice immunized with hMIP-1β gene-modied C126 tumor vaccine could induce tumor specific CTL activity and nonspecific NK activity, and exhibited resistance to later challenge with wild-type C126 cells.Conclusion hMIP-Iβ gene-modified CT26 cells exhibit decreased tumorigenicity, and hMIP-1β genemodified tumor vaccine may induce a powerful specific and nonspecific antitumor response. The data suggested that hMIP-1 β gene-modified tumor vaccine may play a potent role in prevention of metastasis an

关 键 词：人巨噬细胞炎性蛋白-1β 基因修饰 结肠腺癌CT26细胞 抗肿瘤免疫 致瘤性 

分 类 号：R73-36[医药卫生—肿瘤]