GENETIC ENGINEERING OF CANDIDATE VACCINE STRAINS

候选疫苗株的基因工程

• 批准号: 3791038
• 负责人: Bernard Roizman
• 金额: --
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3791038
• 关键词: Herpes simplex disease; Herpesviridae vaccine; attenuated microorganism; cell type; gene deletion mutation; genetic library; genetic manipulation; glycoproteins; herpes simplex virus 1; interleukin 1; laboratory mouse; latent virus infection; microorganism immunology; southern blotting; tissue /cell culture; virulence; virus antigen; virus protein; virus replication

The objectives of this component of the program project grant are several fold: First the project will generate a library of HSV-2 deletion mutants, focusing primarily on several genes previously shown in HSV-1 genomes to affect the capacity of the virus to replicate in CNS tissues. This approach is dictated by the concern that HSV infection can result in encephalitis, with its attendant mortality and morbidity. Studies on HSV-1 have previously shown that deletion mutants incapable of replicating in the CNS of highly susceptible animals can be constructed. At the molecular level, the desirable mutants are those that are avirulent, yet produce vast amounts of virion structural proteins properly displayed on the surfaces of infected cells. Each mutant will be tested in Project 3 for its phenotypic behavior in mice. The second objective of the project is to insert appropriate HSV-1 glycoprotein genes into the suitable candidate vaccine strain(s) selected from the screens described above, in order to enhance immunity against HSV- 1 infection. A number of potential sites for insertion of the HSV-1 genes are available and will be tested. The third objective of this project is to screen the replicative capacity of the deletion mutants in a variety of cell lines and strains, but particularly in cells like the MRC-5 cell strain which has been licensed and approved for viral vaccine production. The fourth objective is to construct and test under appropriate promoters viruses carrying genes that express factors involved in enhancement of mucosal immunity. While such vaccines might well be construed to being in the category of the "next generation", the currently evolving technology of tailoring promoters to required level of expression will be tested using genes specifying particular interleukins. In all of these projects, the design and ultimate structure of the vaccine candidate strains will take cognizance of and rest on the results of studies in Projects 2 and 3.

计划项目授予的该组成部分的目标是几个 折叠： 首先，该项目将产生HSV-2删除突变体的库， 主要专注于先前在HSV-1基因组中显示的基因 影响病毒在中枢神经系统组织中复制的能力。 这 方法是由HSV感染可能导致的关注所决定的 脑炎，其随之而来的死亡率和发病率。 HSV-1的研究 以前已经表明，缺失突变体无法在 可以建造高度易感动物的中枢神经系统。 在分子 级别，理想的突变体是那些无毒的突变体，但产生了广阔的突变体 适当显示在表面上的病毒体结构蛋白量 感染细胞。 每个突变体将在项目3中测试其表型 小鼠的行为。 该项目的第二个目标是插入适当的HSV-1 糖蛋白基因进入了合适的候选疫苗菌株（S） 从上面描述的屏​​幕中，为了增强对HSV-的免疫力 1感染。 许多插入HSV-1基因的潜在位点 可用并将进行测试。 该项目的第三个目标是筛选复制能力 多种细胞系和菌株中的缺失突变体的 特别是在已获得许可的MRC-5细胞应变等细胞中 并批准用于病毒疫苗的生产。 第四个目标是在适当的启动子下构建和测试 携带基因的病毒表达与加强有关的因素 粘膜免疫。 虽然这种疫苗很可能被解释为 “下一代”的类别，目前不断发展的技术 将启动子定制为所需的表达水平，将使用 指定特定白纹素的基因。 在所有这些项目中，疫苗的设计和最终结构 候选菌株将认识并依靠 项目2和3的研究。