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.

项目摘要/摘要 致病病毒的出现和重新出现代表着持续的传染病威胁 公共卫生。其中，副粘病毒，包括许多重要的人和动物 病原体，还包括两个重要的新出现的人畜共患病毒病原体的优秀例子： 尼帕病毒(Nipah Virus，NIV)和亨德拉病毒(Hendra Virus，HEV)。[和合]和戊型肝炎病毒有独特的广泛的宿主取向 能够感染6目哺乳动物中的至少18种动物。[和合]和HEV也可以引起 包括人类在内的11种哺乳动物中的系统性和常常是致命的呼吸系统和/或神经系统疾病。 这些埃尼帕病毒对澳大利亚的人类和经济上重要的牲畜仍然是重要的生物栖息地。 以及整个东南亚。此外，目前还没有批准用于人类使用的疫苗或治疗药物。 海尼帕病毒是单链、负义、包膜的rna病毒，有两层膜。 参与病毒入侵的锚定糖蛋白，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，可促进病毒粒子与宿主细胞膜的融合。这个 病毒G和F糖蛋白是中和抗体的主要抗原靶蛋白，也是病毒的主要焦点 积极的疫苗策略。我们一直在描述非致病物种的海尼帕病毒，雪松病毒 (CedPV)。利用重组病毒糖蛋白介导的细胞融合实验，我们已经确定了CedPV- 其介导的融合趋向性与新城疫病毒和戊型肝炎病毒相似，而CedPV的易福林受体趋向性是 非常广泛的融合是由ewitin-B1和-B2以及糖磷脂酰肌醇- 锚定了A亚型eparins-A1、-A2和-A5。这些伊弗林中的每一个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准备 重组CedPV(RCedPV)我们已经用rCedPV感染研究证实了融合趋向性的发现。 RCedPV平台现在提供了一种新的病毒学系统，可以用于各种应用程序进行研究 在BSL-2遏制下安全地感染埃尼帕病毒，这种病毒将被CETR研究项目和 核心。但更重要的是，它是一个真正的埃尼帕病毒系统，与 NIV和HEV，并可以通过基因操作作为减毒的通用疫苗 平台，以及一个可能能够诱导持久和平衡的免疫反应的平台。使用这一新的 我们的目标是开发编码猪瘟病毒F和G糖蛋白的rCedPV嵌合病毒。 NIV或HEV，并鉴定和探索其潜在的新型海尼帕病毒疫苗。具体来说，我们将：1) RCedPV-NiV/HEV病毒的构建和拯救；2)细胞感染中嵌合病毒的特征 和趋向性研究；3)分析免疫反应性和转归 RCedPVs感染后免疫缺陷小鼠；4)检测rCedPVs的保护能力 在动物受试者中接种疫苗，以应对新城疫和戊型肝炎病毒的挑战。