VEE PATHOGENESIS AND VACCINE DEVELOPMENT

描述（由申请人提供）：申请人是一名具有一定临床和实验室经验的D.V.M.，寻求支持成为病毒发病机制和疫苗开发领域的独立研究人员。他的长期目标是更好地了解人类甲病毒脑炎的发病机制，包括每个病毒基因的作用。这些信息将用于开发更安全、更有效的疫苗和疗法。拟议的研究重点是委内瑞拉马脑炎病毒（VEEV），这是一种重要的自然出现的人类病原体，也是一种强有力的生物武器，被NIAID归类为B类优先病原体。虽然VEEV是一种重要的人类病原体和有效的疫苗表达载体，但对该病毒的非结构蛋白（复制机制）在发病机制中的作用知之甚少。此外，目前还没有获得许可的人VEEV疫苗。目标是：利用大规模高效生产的VEEV/Sindbis嵌合病毒（SINV）开发安全有效的VEEV疫苗；2)利用互反的SINV/ veev嵌合体研究VEEV的发病机制，评估结构蛋白和非结构蛋白在组织趋向性和致病性中的作用。本文将提出两个中心假设：1)VEEV的毒力潜力很大程度上取决于非结构蛋白及其与细胞/有机体因子的相互作用；2) VEEV的组织亲和性不仅取决于病毒结合和进入靶细胞的能力，还取决于病毒建立生产性感染的能力。更好地了解这些VEEV-细胞相互作用将使申请人产生高度减毒的SINV/VEEV嵌合体，从而诱导对VEEV的保护性免疫。具体目标是：1。研究小鼠嵌合SINV/VEEV候选疫苗的体内外复制、致病性和免疫原性。2. 确定相互嵌合的SINV/VEEV病毒在小鼠体内的组织亲和性和致病性。通过该项目，申请人将创造出高度减毒、免疫genie和在细胞培养中高效复制的SINV/VEEV嵌合疫苗候选病毒。通过将VEEV的非结构蛋白与结构蛋白分离，阐明复制机制（非结构蛋白）在其发病机制中的作用。

DESCRIPTION (provided by applicant): The applicant, a D.V.M. with some clinical and laboratory experience, seeks support to become an independent researcher in the field of viral pathogenesis and vaccine development. His long-term goal is to better understand the pathogenesis of human alphaviral encephalitides, including the roles of each viral gene. This information will be used to develop safer and more effective vaccines and therapeutics. The proposed research focuses on Venezuelan equine encephalitis virus (VEEV), an important, naturally emerging human pathogen and a potent biological weapon categorized by the NIAID as a Category B Priority Pathogen. Although VEEV is an important human pathogen and an efficient vaccine expression vector, little is known about the role of the nonstructural proteins (replicative machinery) of this virus in pathogenesis. Furthermore, no licensed human VEEV vaccine is available. The goals are to1l) develop safe and effective VEEV vaccines using chimeric VEEV/Sindbis viruses (SINV) that are efficiently produced at large scale, and; 2) study VEEV pathogenesis using reciprocal SINV/VEEVchimeras to assess the roles of structural vs. nonstructural proteins in tissue tropism and pathogenicity. Two central hypotheses will be addressed: 1) The virulence potential of VEEV strongly depends on the nonstructural proteins and probably their interactions with cellular/organismal factors, and; 2) VEEV tissue tropism depends on the ability of the virus not only to bind and enter target cells, but also to establish productive infection. Better understanding of these VEEV-cell interactions will allow the applicant to generate highly attenuated SINV/VEEV chimeras that induce protective immunity against VEEV. The specific aims are: 1. Characterize the replication in vitro and in vivo, pathogenicity and immunogenicity of chimeric SINV/VEEV vaccine candidates in mice. 2. Determine the tissue tropism and pathogenicity of reciprocal, chimeric SINV/VEEV viruses in mice. Through this project, the applicant will create SINV/VEEV chimeric vaccine candidate viruses that are highly attenuated, immunogenie and replicate efficiently in cell culture. By dissecting the nonstructural proteins of VEEV from the structural proteins, the role of replieative machinery (nonstructural proteins) in the pathogenesis will be elucidated.