VEE PATHOGENESIS AND VACCINE DEVELOPMENT

描述（由申请人提供）：申请人，D.V.M.。具有一定的临床和实验室经验，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性脑炎的发病机制，包括每个病毒基因的作用。这些信息将用于开发更安全，更有效的疫苗和治疗方法。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NIAID归类为B类优先病原体的强大生物武器。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，没有获得许可的人VEEV疫苗可用。目的是1）使用大规模有效生产的嵌合VEEV/辛德毕斯病毒（SINV）开发安全有效的VEEV疫苗; 2）使用相互的SINV/VEEV嵌合体研究VEEV发病机制，以评估结构蛋白与非结构蛋白在组织嗜性和致病性中的作用。将讨论两个中心假设：1）VEEV的毒力潜力强烈依赖于非结构蛋白，可能是它们与细胞/生物因子的相互作用; 2）VEEV组织嗜性不仅取决于病毒结合和进入靶细胞的能力，还取决于建立生产性感染的能力。更好地理解这些VEEV-细胞相互作用将允许申请人产生高度减毒的SINV/VEEV嵌合体，其诱导针对VEEV的保护性免疫。具体目标是：1.表征嵌合SINV/VEEV疫苗候选物在小鼠中的体外和体内复制、致病性和免疫原性。2.确定相互嵌合的SINV/VEEV病毒在小鼠中的组织嗜性和致病性。通过该项目，申请人将创建高度减毒、免疫原性和在细胞培养中有效复制的SINV/VEEV嵌合疫苗候选病毒。通过将VEEV的非结构蛋白从结构蛋白中分离出来，阐明了复制机制（非结构蛋白）在VEEV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.