项目名称: 猪SLA I分子超级型及猪源重要病毒CTL交叉表位的研究
项目编号: No.31201887
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: 张念之
作者单位: 中国农业大学
项目金额: 23万元
中文摘要: 对猪主要组织相容性复合体(SLA)I分子进行超级型分类,鉴定猪源病毒的细胞毒性T淋巴细胞(CTL)交叉表位是开发猪用CTL表位疫苗的基础,对于控制猪源病毒性疾病有重要的价值。本项目拟大量克隆、表达我国猪群中存在的SLA I分子,对其进行精确的分子建模;在此基础上对不同SLA I分子的抗原结合区域进行分析,根据氨基酸组成的差异,对我国猪群中主要的SLA I分子进行超级型分类;利用生物信息学手段,筛选表型覆盖率高的SLA I超级型分子所能结合的猪源重要病毒(猪瘟、蓝耳病、口蹄疫、猪流感)的CTL表位;人工合成表位多肽,利用体外共复性、圆二色谱等技术鉴定多肽与相应SLA I分子的结合力,验证SLA I超级型分类的正确性;进一步利用四聚体、流失细胞术及酶联免疫斑点实验等技术,鉴定结合力强的多肽是否为功能性CTL交叉表位。研究结果将为解决CTL表位疫苗在猪群中适用范围窄的难题提供理论支持和数据储备。
中文关键词: 猪白细胞受体;超级型;晶体结构;CTL表位;猪源病毒
英文摘要: Developing swine major histocompatibility complex (also named swine lymphocyte antigen, SLA) class I supertype system and indentifying CTL cross-reaction epitopes of important swine-origin viruses are essential for the design of porcine CTL epitope vaccine and control the swine-origin zoonoses. In this study, the SLA I molecules of domestic pigs will be massively cloned and expressed, and their 3D models will be constructed to analyze their antigen-binding domains. According to the amino acid compositions of the antigen-binding grooves, the SLA I molecules will be classified into different supertypes. The predominant SLA supertypes covering great percent of domestic pig will be identified. The CTL epitopes from important swine-origin virues (Classical swine fever virus,CSFV;Porcine reproductive and respiratory syndrome virus, PRRSV; Foot and mouth disease virus, FMDV; Swine-origin influenza virus, S-OIV) binding with major SLA I supertype molecules will be screened by using bioinformatics methods. The selected peptides will be synthesized, and their binding affinities with corresponding SLA I molecules will be checked by co-refolding and circular dichroism spectrum (CD) in order to test the SLA I supertype classification. At last, the functional CTL cross-reaction epitope peptides will be identified by SLA I tet
英文关键词: Swine lymphocyte antigen;Supertype;crystal structure;CTL epitope;Swine-origin Virus