鱼类弹状病毒重组多表位肽的构建及免疫效应研究

项目名称： 鱼类弹状病毒重组多表位肽的构建及免疫效应研究

项目编号： No.31202028

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 陈中元

作者单位： 中国科学院水生生物研究所

项目金额： 25万元

中文摘要： 弹状病毒是危害鱼类及多种水生动物的烈性、传染性病毒病原。申请者在已获得鳜鱼弹状病毒全基因组序列及多株单克隆抗体、以及对病毒免疫相关蛋白的抗原性进行初步分析的基础上，拟进行鱼类弹状病毒重组多表位肽的构建及免疫效应研究。利用单抗、生物信息学、肽扫描技术等对鳜鱼弹状病毒糖蛋白（G）与核蛋白（N）蛋白的T、B细胞抗原表位进行预测和筛选；然后将候选抗原表位基因经不同组合、串联，表位之间以柔性短肽进行间隔，获得多表位肽编码基因片段，分别构建原核与真核表达系统；进一步以纯化的多表位融合蛋白或重组真核表达质粒免疫鱼体，经攻毒实验测试不同多表位肽的免疫抗病效果，并从鱼体特异性抗体的效价、淋巴细胞增殖情况、以及免疫相关基因表达变化等方面，了解其诱导宿主的免疫应答作用，揭示免疫效应机制。通过上述研究，以期获得免疫原性强、可诱导高效免疫保护效果的多表位肽，从而为鱼类弹状病毒新型高效疫苗的研发提供候选抗原。

中文关键词： 鳜鱼弹状病毒；糖蛋白；核蛋白；多表位肽；免疫反应

英文摘要： Rhabdovirus is a highly infectious viral pathogen of fish and other aquatic animals. Based on the complete genome sequence and monoclonal antibodies of Siniperca chuatsi rhabdovirus (SCRV), as well as the antigenic analysis of viral proteins, we will construct recombinant multi-epitope peptides of fish rhabdovirus, and investigate their effects on the antiviral immune responses in mandarin fish. The B cell epitopes of SCRV G protein will be identified by using monoclonal antibodies with pepscan, and the T cell epitopes will be analyzed by epitope prediction programs. The multi-epitope peptides coding gene fragments will be obtained with recombinant DNA technology, and then subcloned into the prokaryotic expression vector, or into the eukaryotic expression vector to construct multi-epitope DNA vaccines against SCRV. The protective effects of different multi-epitope peptides will be analyzed and compared in mandarin fish vaccinated with the purified recombinant proteins or multi-epitope DNA vaccines, and the peptides offering good protection would be screened out. In order to understand the antiviral immune responses elicited by multi-epitope peptides, we will further analyze the virus-specific antibody titers, the proliferation of lymphocytes, and the expression patterns of immune related genes in vaccinated fish

英文关键词： Siniperca chuatsi rhabdovirus (SCRV)；glycoprotein；nucleoprotein；multi-epitope peptides；immune response