Alphaviral Determenants of Infection in Mice and Vectors

摘要：东部、委内瑞拉和西部马脑炎(EEE、VEE、WEE)病毒(甲型病毒； 霍乱弧菌(Togaviridae)是由蚊子传播的病毒，会导致人和马的严重脑炎。三个都是 病毒被NIAID和CDC列为B类病原体，并被列为潜在的生物恐怖主义/生物武器 (BT/BW)代理。在传播潜力、蚊子宿主等方面存在巨大的知识差距 决定因素、气雾化病毒的致病机制、获得性和先天免疫反应等 病毒群，尤指EEEV和WeEV。在这个项目中，我们将开发甲型病毒(EEEV和WeEV) 感染性克隆用于确定病毒复制的分子机制及其在小鼠体内的致病作用 以及在Bt/BW事件期间可能作为二次传播媒介的蚊子的传染性。 我们还将开发稳定的、双亚基因组EEEV、VEEV和WeEV转导病毒以 研究它们在受感染的小鼠和载体中复制和表达报告基因的能力。我们还将 在小动物模型上进行主要集中于EEEV和WeEV的发病机制研究，但也将 在这些研究中包括从三种甲型病毒中转导病毒。我们将重点研究其发病机制。 小鼠通过媒介和气雾剂感染途径感染。最后，我们将描述病毒载体的特征 三种病毒的相互作用可能导致Bt/BW载体传播潜力增加 病原体和新出现的甲型病毒病原体。 项目互动：项目的PI是安·M·鲍尔斯博士和肯·E·奥尔森博士。鲍尔斯博士和 奥尔森位于疾病控制中心，媒介传播传染病和 科林斯堡的节肢动物传播传染病实验室(CSU)。鲍尔斯博士和 奥尔森两人都在甲型病毒生物学、分子生物学、媒介生物学方面拥有丰富的经验，并 发展甲型病毒与小动物模型相互作用方面的专业知识。此外，该项目还将与 与RCE中的其他调查人员。具体地说，与Kedl、Blair和Dow博士一起进行的Lanacs研究(II.C.6)， 提图斯和比蒂博士(II.B.2.)和张博士(II B.3)关于新的疫苗技术，Morrey博士(II G)在 共享在该项目中开发的传染性甲型病毒克隆和转导病毒，Fujinami博士和 FOY(II.H.2)关于基于RNAi的抗病毒疗法。该项目将成为开发协议的资源， 病毒，以及可被RCE其他成员使用的试剂。最后，这个项目将依赖于 关于两个证监会：动物模型核心(III.A)和基因组/蛋白质组学核心(III.C)。

Abstract: Eastern, Venezuelan, and Western equine encephalitis (EEE, VEE, WEE) viruses (Alphavirus; Togaviridae) are mosquito-borne viruses causing severe encephalitis in humans and horses. All three viruses are listed as category B agents by both NIAID and CDC and as potential bioterrorism/biowarfare (BT/BW) agents. There are enormous knowledge gaps concerning transmission potential, mosquito-host determinants, pathogenesis from aerosolized virus, acquired and innate immune responses, etc. for this group of viruses, especially EEEV and WEEV. In this project, we will develop alphavirus (EEEV and WEEV) infectious cDNA clones to determine molecular mechanisms of viral replication and pathogenesis in mice and infectivity of mosquitoes that may serve as vectors in secondary transmission during a BT/BW event. We will also develop stable, double subgenomic EEEV, VEEV, and WEEV transducing viruses to characterize their ability to replicate and express a reporter gene in infected mice and vectors. We will also conduct pathogenesis studies focusing primarily on EEEV and WEEV in small animal models, but will also include transducing viruses from the three alphaviruses in these studies. We will focus on pathogenesis in mice following infection by vector and aerosol routes of infection. Finally we will characterize virus-vector interactions for all three viruses that may lead to increased transmission potential by vectors of BT/BW agents and emerging alphavirus pathogens. Project Interactions: The Pi's of the project are Drs. Ann M. Powers and Ken E. Olson. Drs. Powers and Olson are located at the Centers for Disease Control, Division of Vector-borne Infectious Diseases and Arthropod-borne Infectious Diseases Laboratory (CSU) in Fort Collins, Co., respectively. Dr. Powers and Olson both have considerable experience in alphavirus biology, molecular biology, vector biology and are developing expertise in alphavirus interactions with small animal models. In addition, the project will interact with other investigators in the RCE. Specifically, with Drs. Kedl, Blair, and Dow on Lanacs studies (II.C.6), Drs. Titus and Beaty (II.B.2.) and Dr. Chang (II.B.3) on novel vaccine technologies, Dr. Morrey (II.G) in sharing infectious alphavirus clones and transducing viruses developed in this project and Dr. Fujinami and Foy (II.H.2) on RNAi-based antiviral therapies. This project will be a resource for developing protocols, viruses, and reagents that can be used by other members of the RCE. Finally, this project will be dependent on two SFC's: Animal Models Core (III.A) and Genomics/Proteomics Core (III.C).