Emerging Viruses

已知在人类中引起具有高发病率和死亡率的出血热疾病的病毒剂，例如 布尼亚病毒-克里米亚-刚果出血热病毒、裂谷热病毒和汉坦病毒或丝状病毒- 埃博拉和马尔堡病毒是生物防御关注的优先病原体。此外，还发生了几起疫情 新发现和/或新出现的病毒，这些病毒也导致了大量的人类死亡， 这些病毒包括西尼罗黄病毒和尼帕和亨德拉亨尼帕病毒。这些新出现的病毒 也被归类为生物防御剂，因为它们具有使它们适合的特性 用于武器化对于所有这些病毒，既没有批准的疫苗，也没有治疗方法来预防 感染或疾病。作为一种潜在的填补空白的方法，快速开发安全有效的抗病毒药物， 针对这些药物的治疗，我们建议鉴定、分离和表征中和性人单克隆抗体， 在一些实施方案中，抗体是与这些病毒的包膜糖蛋白（Env）的天然形式反应的抗体（nhMAb）。发展这些 nhMAb将提供一组有价值的暴露后或感染后治疗剂，以对抗由以下引起的疾病： 这些代理人。抗体将通过针对病毒Env筛选人噬菌体展示文库来获得。我们 研究项目将由四个子项目组成。这些项目的统一战略将是发展 用于测量病毒进入沿着天然病毒Env的产生的基于重组的测定法， 噬菌体淘选程序。子项目1将开发亨尼帕病毒的检测和Env抗原， 布尼亚病毒;子项目2将侧重于黄病毒;子项目3将采用埃博拉病毒和马尔堡病毒 子项目4将开发第二代基于甲病毒的复制子系统作为疫苗平台 利用来自其他子项目的一个或多个病毒Env。研究的总体目标 项目将开发一系列有效的nhMAb，能够用作针对多种病毒的被动免疫疗法 重要的病毒病原体，并开发能够引发体液应答的疫苗平台，其包括 其他子项目开发的nhMAb观察到的抗体反应性。整个研究项目 具体目标是：1）开发用于病毒进入的高通量重组检测方法，并开发重组病毒 2）通过使用重组病毒Envs鉴定和分离能够阻断病毒进入的Fab和scFv和/或 作为噬菌体淘选抗原的功能性假型化或病毒样颗粒;和3）开发和测试一种新的 基于甲病毒的疫苗平台。总之，本研究计划的短期目标将是开发 nhMAbs可用于被动免疫治疗，而长期目标是创造疫苗 在一个实施方案中，本发明提供了能够在动物宿主中引发类似抗体应答的平台。这两个研究目标都是优先考虑的， 病毒性出血热和新出现病毒生物防御研究的近期和长期目标。

Viral agents known to cause hemorrhagic fever disease with high morbidity and mortality in humans, such as the Bunyaviruses - Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, and Hantaan virus or the Filoviruses - Ebola and Marburg virus, are priority pathogens of biodefense concern. In addition, there have been several outbreaks of newly discovered and/or emerging viruses which have also resulted in significant numbers of human fatalities, and these include the Flavivirus West Nile virus and the Henipaviruses Nipah and Hendra virus. These emerging viruses have also been classified as agents of biodefense concern because they possess characteristics making them suitable for weaponization. For all of these viruses there are neither approved vaccines nor therapeutics for the prevention of infection or disease. As a potential gap-filling approach for the rapid development of safe and efficacious antiviral therapies for these agents, we propose to identify, isolate and characterize neutralizing human monoclonal antibodies (nhMAbs) reactive to the native forms of the envelope glycoproteins (Envs) of these viruses. The development of these nhMAbs would provide a valuable battery of post-exposure or post-infection therapeutics to combat disease caused by these agents. The antibodies will be obtained by screening human phage display libraries against the viral Envs. Our Research Project will be comprised of four Sub-Projects. The unifying strategy of these projects will be the development of recombinant-based assays for measuring virus entry along with the production of native viral Envs as antigens for phage panning procedures. Sub-Project 1 will develop the assays and Env antigens for the Henipaviruses and Bunyaviruses; Sub-Project 2 will focus on the Flaviviruses; Sub-Project 3 will employ the Ebola and Marburg virus systems; while Sub-Project 4 will develop a second-generation alphavirus-based replicon system as a vaccine platform utilizing one or more of the viral Envs derived from the other Sub-Projects. The overall objectives of the Research Project will be to develop a battery of potent nhMAbs capable of being used as passive immunotherapy against several important viral pathogens, and develop a vaccine platform capable of eliciting a humoral response which includes the antibody reactivities observed by those nhMAbs developed by the other Sub-Projects. The overall Research Project's Specific Aims will be 1) Develop high-throughput recombinant assays for virus entry and develop recombinant viral Envs; 2) identify and isolate Fabs and scFvs capable of blocking virus entry by using recombinant viral Envs and/or functional pseudotyped or virus-like particles as antigens for phage panning; and 3) develop and test a novel alphavirus-based vaccine platform. In summary, the short-term goals of this research Program will be the development of nhMAbs that could be used for passive immunotherapy, while the long-term objective is the creation of a vaccine platform capable of eliciting similar antibody responses in an animal host. Both of these research goals are priorities as immediate and long-term objectives for biodefense research on viral hemorrha0ic fevers and emerging viruses.