转人B7-H3基因鳞癌细胞瘤苗体外诱导抗肿瘤免疫应答的研究

Antitumor Immune Responses Induced by Gene Transfer of B7-H3 into Squamous Cell Carcinoma Tca8113 in Vitro

目的:研究转人B7-H3基因鳞癌细胞疫苗体外诱导抗肿瘤免疫应答的能力。方法:采用脂质体介导法将真核表达质粒pEGFP-B7-H3导入人鳞癌细胞Tca8113,经G418筛选后获得稳定高表达克隆,以丝裂霉素C(MMC)处理后,制成肿瘤细胞疫苗,经体外与人外周血淋巴细胞共同培养后,测定淋巴细胞特异性杀伤活性及对淋巴细胞产生细胞因子的影响。结果:转染B7-H3的Tca8113细胞能够高表达B7-H3蛋白。经MMC处理后,与野生型的Tca8113细胞相比,上述瘤苗能诱导淋巴细胞产生针对Tca8113的特异性杀伤作用,能显著增强淋巴细胞分泌IFN-γ的能力。结论:转B7-H3基因人鳞癌细胞疫苗能诱导有效的抗鳞癌免疫反应。

Objective： To study the antitumor immune responses induced in vitro by human B7 - H3 gene transfected Tca8113. Methods： The eukaryotic expression vector pEGFP - B7 - H3 was transfected into human squamous cell carcinoma cell line Tca8113 by Lipofectamine. Then the transfected cells were selected in medium containing G418 （600μg/ml） and termed as Tca8113 - hB7 - H3. The TCV - hB7 - H3 was obtained by treating them with mitomycin（MMC） . After co -culture TCV with human lymphocytes , the lymphocytes were tested for cytotoxicity against TcaS113 cells and the supernatants were harvested to detect the cytokines （IFN-γ）. Results： Tca8113 -hB7 -H3 cells expressed B7 -H3 protein with the highest cell surface level. After treated with MMC （50ug/ml） ,the tumor cell vaccine derived from Tca8113 -hB7 - H3 （ TCV - hB7 - H3 ） could induce a more efficient cytotoxic activity of human lymphocyte against its parental tumor cell Tea8113 compared with TCV -Tca8113. Higher levels of IFN - γ were released by the spleenocytes after stimulated by TCV - hB7 - H3. Conclusion ： These results suggest the expression of hB7 - H3 by squamous carcinoma cells is effective ton induce antitumor immune responses.