4-1BBL修饰小鼠结肠癌疫苗体外诱导抗肿瘤免疫反应的研究

In Vitro Anti-tumor Immune Effects Induced by 4-1BBL-transfected Mouse Colon Cancer Cells

背景与目的:研究4-1BBL(4-1BB Ligand,即CD137配体)转基因小鼠结肠癌细胞瘤苗体外诱导细胞毒性T淋巴细胞(CytotoxicTlymphocytes,CTL)特异性杀伤活性及刺激淋巴细胞产生细胞因子的作用。材料与方法:采用脂质体介导法将真核表达质粒pMKITneo/4-1BBL导入小鼠结肠癌colon26细胞,经G418筛选后获得4-1BBL高表达克隆,用丝裂霉素C(MMC)处理后,制成肿瘤细胞瘤苗,与经体外诱导的同系小鼠CTL共同培养,测定对CTL特异性杀伤活性及对脾细胞产生细胞因子(IL-2、IL-4和IFN-γ)的影响。结果:转染4-1BBL的colon26细胞高表达4-1BBL蛋白,将该细胞经MMC处理后制成的瘤苗,与野生型colon26细胞相比,对CTL特异性杀伤亲本肿瘤细胞的作用明显增强(P<0.01),但CTL对非亲本肿瘤细胞的杀伤作用无明显影响(P>0.05);该瘤苗在体外能显著增强脾细胞分泌细胞因子(IL-2、IL-4和IFN-γ)的能力(P<0.01)。结论:4-1BBL转基因小鼠结肠癌细胞瘤苗能有效诱导抗结肠癌免疫反应。

BACKGROUND ＆ AIM： To study the cytotoxic activity of CTL and cytokines production of lymphocytes induced by 4-1BBL-transfected colon26 in vitro. MATERIALS AND METHODS： The eukaryotic expression vector pMKIT^neo/ 4-1BBL was transfected into colon26 by lipofectamine^2000, then the cells with high expression of 4-1BBL were selected by G418. The tumor cell vaccines （TCV） were obtained by treatment with mitomycin（MMC） .The TCV were co-cultivated with syngeneic murine cytotoxic T lymphocytes （CTL）.Then the cytotoxic activity of CTL and the cytokines production of spleen cells were tested. RESULTS： The colon26 cells transfected with 4-1BBL showed strong expression of 4-1BBL protein on cell surface. Compared with TCV-colon26, the TCV-colon26/ 4-1BBL cells could induce a more efficient cytotoxic activity of CTL against its parental tumor cell colon26（P〈0.01）, but not against non-parental tumor cell S180.The TCV-colon26/ 4-1BBL could significantly enhance the ability of murine spleen cells to produce cytokines（P〈0.01） CONCLUSION： The mufine colon26 tumor cells transfected with 4-1BBL could induce an efficient anti-tumor immune response.