基于肿瘤抗原PBK的融合蛋白疫苗的设计制备及抗肿瘤活性  

Design,preparation,and antitumor activity of fusion protein vaccine based on tumor antigen PBK

作　　者：钱茜茜 李国志 田浤[1] 高向东[1] QIAN Qianqian;LI Guozhi;TIAN Hong;GAO Xiangdong(Jiangsu Provincial Key Laboratory of Druggability of Biopharmaceuticals,School of Life Science and Technology,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学生命科学与技术学院、江苏省生物药物成药性研究重点实验室,南京211198

出　　处：《中国药科大学学报》2024年第5期657-665,共9页Journal of China Pharmaceutical University

基　　金：新疆维吾尔自治区重点研发任务专项(2020B03003)。

摘　　要：PBK是一种癌症/睾丸抗原,其在人类的多种正常组织中不表达,而在癌变后的组织细胞中异常过表达,促进癌症的发生、转移甚至耐药性,为肿瘤免疫治疗提供了新靶点。本研究将硝基化T细胞表位与PBK蛋白融合表达,构建了以PBK为靶点的蛋白疫苗PBK-Nitrath;采用IFN-γ ELISpot法评估免疫小鼠脾脏中PBK抗原特异性T细胞的激活水平,并在体外进行细胞毒性T细胞杀伤效应检测评估其对H22肝癌细胞的杀伤能力;在此基础上采用H22肝癌皮下移植瘤模型对其抗肿瘤活性进行评价,并通过流式细胞术对外周血和脾脏T淋巴细胞的分化情况与肿瘤的免疫浸润情况进行分析。结果显示,PBK-Nitrath能够显著地诱导PBK抗原特异性T细胞的激活,增强细胞毒性T淋巴细胞的杀伤能力,并可显著抑制小鼠肝癌肿瘤生长,提高外周血与脾脏中CD8~+CD107a~+T细胞的比例,同时可促进肿瘤淋巴细胞的浸润。研究结果提示PBK-Nitrath是有潜力的肿瘤疫苗候选分子。PBK is a cancer/testis antigen that exhibits absent expression in various normal human tissues,but undergoes aberrant overexpression upon cellular transformation,thereby promoting the initiation,metastasis and even drug resistance of cancer.Consequently,it represents a novel target for tumor immunotherapy.In this study,PBK-Nitrath,a protein vaccine specifically designed to target PBK by fusing nitrated T epitope with the PBK protein was developed,using the IFN-γELISpot method to evaluate the activation level of PBK antigen-specific T cells in the spleen of immunized mice,and conducting in vitro cytotoxicity T cell killing efficacy test to evaluate the killing ability against H22 hepatic carcinoma cells;the anti-tumor activity was evaluated using a H22 hepatic carcinoma subcutaneous transplantation tumor model,and the differentiation of peripheral blood and spleen T lymphocytes and tumor immune infiltration were analyzed by flow cytometry.Results showed that PBK-Nitrath could efficiently induce the activation of antigen-specific T cells against PBK while enhancing cytotoxic T lymphocyte-mediated killing capacity,significantly impede hepatic carcinoma progression in mice and increase the ratio of CD8+CD107a+T cells within peripheral blood and spleen,and facilitate tumor lymphocyte infiltration.Our findings reveal the potential utility of PBK-Nitrath as an effective candidate for tumor vaccine.

关 键 词：PBK 肿瘤免疫 肿瘤疫苗 肝癌 

分 类 号：R186[医药卫生—流行病学] Q816[医药卫生—公共卫生与预防医学]