Project 3 - USUHS

项目3-USUHS

• 批准号: 10581504
• 负责人: CHRISTOPHER C BRODER
• 金额: $ 63.27万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-20 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581504
• 关键词: Animals; Antibodies; Attenuated; Australia; B-Lymphocytes; Bangladesh; Binding; Biological Assay; Cell Communication; Cell fusion; Cell membrane; Cells; Cellular biology; Chimera organism; Chimeric Proteins; Communicable Diseases; Containment; Eph Family Receptors; Ephrin-A1; Ephrin-B1; Ephrins; Far East; Ferrets; Future; Glycoproteins; Growth; Hamsters; Hendra Virus; Henipavirus; Human; Immune response; Immunization; Immunocompetent; Immunodeficient Mouse; Immunologic Deficiency Syndromes; Infection; Kinetics; Livestock; Luc Gene; Luciferases; Mammals; Mediating; Medical; Membrane; Membrane Fusion; Modeling; Mus; Nipah Virus; Outcome; Paramyxovirus; Pathogenesis; Pathogenicity; Persons; Public Health; RNA Viruses; Reagent; Receptor Cell; Recombinants; Research Project Grants; Respiratory Disease; Series; System; T-Lymphocyte; Technology; Testing; Texas; Therapeutic; Tropism; Universities; Vaccination; Vaccines; Viral; Virion; Virus; Virus Diseases; Zoonoses; biothreat; experimental study; genetic manipulation; glycoprotein G; in vivo; innovation; nervous system disorder; neutralizing antibody; nonhuman primate; novel; pathogen; pathogenic virus; receptor; recombinant virus; response; reverse genetics; transmission process; vaccine platform; vaccine strategy; virus envelope

Project Summary/Abstract The emergence and reemergence of pathogenic viruses represent continuous infectious disease threats to public health. Among these, the paramyxoviruses, which include many important human and animal pathogens, also include two excellent examples of emerged, zoonotic viral pathogens of importance: the henipaviruses; Nipah virus (NiV) and Hendra virus (HeV). NiV and HeV have a uniquely broad host tropism capable of infecting at least 18 animal species across 6 orders of mammals. NiV and HeV can also cause a systemic and often fatal respiratory and/or neurological disease in 11 mammalian species including humans. These henipaviruses remain significant biothreats to humans and economically important livestock in Australia and throughout South East Asia. In addition, there are no vaccines or therapeutics approved for human use. The henipaviruses are single-stranded, negative sense, enveloped RNA viruses and possess two membrane anchored glycoproteins involved in virus entry, one mediates host cell receptor attachment (G glycoprotein) and the other is a Class I fusion (F) glycoprotein which facilitates virion and host cell membrane fusion. The viral G and F glycoproteins are the major antigenic targets of neutralizing antibodies and are the main focus of active vaccine strategies. We have been characterizing the non-pathogenic species of henipavirus, Cedar virus (CedPV). Using recombinant viral glycoprotein mediated cell fusion assays, we have determined that CedPV- mediated fusion tropism is similar to NiV and HeV, however, the ephrin receptor tropism of CedPV is remarkably broad and fusion is triggered by ephrin-B1 and -B2 as well as the glycophosphatidylinositol- anchored A subtype ephrins-A1, -A2, and -A5. The fusion tropism activity by each of these ephrins is also correlated with CedPV G binding, and using a recently developed reverse genetics system for preparing recombinant CedPV (rCedPV) we have confirmed the fusion tropism findings with rCedPV infection studies. The rCedPV platform now offers a new virological system that can be used in a variety of applications to study henipaviruses safely under BSL-2 containment which will be broadly used by the CETR Research Projects and Cores. But of additional importance, it is an authentic henipavirus system that is attenuated in comparison to NiV and HeV, and can be genetically manipulated to serve as a live-attenuated universal henipavirus vaccine platform, and one likely capable of inducting a long-lasting and balanced immune response. Using this new system our objectives here will be to develop rCedPV chimeric viruses encoding the F and G glycoproteins of NiV or HeV and characterize and explore their potential a novel henipavirus vaccine. Specifically, we will: 1) Construct and rescue rCedPV-NiV/HeV viruses; 2) Characterize the chimeric viruses in cell-based infection and tropism studies; 3) Analyze the immune responses and outcomes in both immunocompetent and immunodeficient mice following infection with rCedPVs; 4) Test the protective abilities of rCedPVs by vaccination in animal subjects, in response to NiV and HeV challenge.

项目概要/摘要 致病病毒的出现和再次出现代表着对人类的持续传染病威胁 公共卫生。其中，副粘病毒包括许多重要的人类和动物病毒 病原体，还包括出现的两个重要的人畜共患病毒病原体的绝佳例子： 亨尼帕病毒；尼帕病毒（NiV）和亨德拉病毒（HeV）。 NiV 和 HeV 具有独特的广泛宿主向性 能够感染 6 个目哺乳动物中至少 18 种动物。 NiV 和 HeV 也会导致 包括人类在内的 11 种哺乳动物的系统性且通常致命的呼吸系统和/或神经系统疾病。 这些亨尼帕病毒仍然对澳大利亚人类和经济上重要的牲畜构成重大生物威胁 以及整个东南亚。此外，还没有批准用于人类使用的疫苗或治疗方法。 亨尼帕病毒是单链、负义、有包膜 RNA 病毒，具有两个膜 参与病毒进入的锚定糖蛋白，一种介导宿主细胞受体附着（G糖蛋白） 另一种是I类融合（F）糖蛋白，促进病毒粒子和宿主细胞膜融合。这 病毒G和F糖蛋白是中和抗体的主要抗原靶点，也是研究的主要焦点。 积极的疫苗策略。我们一直在表征亨尼帕病毒、雪松病毒的非致病性物种 （CedPV）。使用重组病毒糖蛋白介导的细胞融合测定，我们确定 CedPV- 介导的融合向性与 NiV 和 HeV 相似，然而，CedPV 的肝配蛋白受体向性是 非常广泛，融合由肝配蛋白-B1和-B2以及糖磷脂酰肌醇-触发 锚定 A 亚型肝配蛋白 -A1、-A2 和 -A5。这些肝配蛋白的融合趋向性活性也是 与 CedPV G 结合相关，并使用最近开发的反向遗传学系统来制备 重组 CedPV (rCedPV) 我们已经通过 rCedPV 感染研究证实了融合向性发现。 rCedPV 平台现在提供了一种新的病毒学系统，可用于多种研究应用 亨尼帕病毒在 BSL-2 遏制下安全，将被 CETR 研究项目广泛使用 核心。但更重要的是，它是一个真正的亨尼帕病毒系统，与 NiV 和 HeV，可通过基因操作作为通用亨尼帕病毒减毒活疫苗 一个可能能够诱导持久且平衡的免疫反应的平台。使用这个新的 我们的目标是开发 rCedPV 嵌合病毒，编码 F 和 G 糖蛋白 NiV 或 HeV，并表征和探索它们作为新型亨尼帕病毒疫苗的潜力。具体来说，我们将：1） 构建并拯救 rCedPV-NiV/HeV 病毒； 2) 表征嵌合病毒在细胞感染中的作用 和向性研究； 3) 分析免疫能力强和免疫能力强的人的免疫反应和结果 感染 rCedPV 后免疫缺陷的小鼠； 4) 通过以下方式测试 rCedPV 的保护能力 动物受试者接种疫苗，以应对 NiV 和 HeV 攻击。