Single-cycle Infectious Candid#1 to Study Hemorrhagic Fever New World Arenavirus

单周期传染性坦诚

• 批准号: 8385029
• 负责人: Luis Martinez-Sobrido
• 金额: $ 7.68万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8385029
• 关键词: Adverse effects; Antiviral Agents; Arenavirus; Arenavirus Infections; Attenuated Vaccines; Biological; Biological Assay; Biology; Bioterrorism; Cell Line; Cells; Detection; Development; Disease; Drug Delivery Systems; Family; Generations; Genetic; Genome; Glycoproteins; Goals; Green Fluorescent Proteins; Human; Infection; Junin virus; Label; Laboratories; Lassa virus; Lead; Licensing; Life; Life Cycle Stages; Lymphocytic choriomeningitis virus; Mediating; Morbidity - disease rate; Old World Arenaviruses; Plasmids; Prevention; Property; Public Health; RNA; Reagent; Recombinants; Reporter; Reporter Genes; Research; Research Personnel; Ribavirin; Screening procedure; System; Tacaribe Complex Viruses; Technology; Therapeutic; Time; Vaccines; Viral; Viral Hemorrhagic Fevers; Virus; Virus Inhibitors; Virus Replication; Work; base; biodefense; biosafety level 2 facility; drug discovery; high throughput screening; inhibitor/antagonist; innovation; member; mortality; neutralizing antibody; nucleoside analog; pathogen; positional cloning; prophylactic; receptor; stable cell line; vaccine candidate; vaccine development; vector; viral detection; virus identification

DESCRIPTION (provided by applicant): Arenaviruses include several causative agents of Hemorrhagic Fever (HF) disease in humans that are associated with high morbidity and significant mortality. Moreover, weaponized forms of arenaviruses pose a serious threat as agents of bioterrorism. Public health concerns posed by arenaviruses are aggravated by the lack of licensed vaccines and by current anti-arenavirus therapy limited to the off-label use of ribavirin, which is only partially effective and often associated with severe side effects. Limitations in the study of HF arena-viruses include the manipulation of live forms of these agents under BioSafety Level (BSL) 4 laboratories and the requirement of secondary assays for detection of the virus. Development of valid single-cycle infectious surrogates that allow the study of HF arenavirus under less-strict BSL2 facilities and that express a reporter gene for easy viral detection will facilitate the identification of prophylactic and therapeutic strategies using safe, sensitive and specific screening assays compatible with High Throughput Screening (HTS) technologies. Furthermore, they will also represent promising vaccine approaches that circumvent concerns about potential out-of-control replication of attenuated vaccine strains that might lead to disease. Recently, we have described, for the first time, the generation of a single-cycle infectious, reporter-expressing, Old World Lymphocytic Choriomeningitis Virus (LCMV) where we replaced the viral glycoprotein (GP) with a reporter green fluorescent protein (GFP), rLCMV?GP/GFP. Infectious virus was achieved via genetic trans- complementation with cell lines constitutively expressing LCMV GP. This system allowed us to study multiple aspects of the virus and to develop screening assays for detection and quantification of neutralizing antibodies and identification of viral inhibitors. We were also able to extend our work to Lassa virus (LASV), an HF member of the Old World arenavirus, by generating LASV GP-expressing cell lines and LASV GP-pseudotyped rLCMV?GP/GFP. Since Old World and New World arena-viruses differ in their reservoirs, cellular receptor use for viral entry, and RNA genome composition, the development of valid surrogates to study New World arenavirus is imperative to provide researchers a way to study all aspects of the virus biology, to identify antivirals, and to develop vaccines against New World arenaviruses under less strict biosafety laboratories. In this application we propose to expand our recently described technology to the study of New World arenavirus by generating a single-cycle infectious, reporter-expressing, attenuated vaccine strain Candid#1 (rCan?GP/GFP). PUBLIC HEALTH RELEVANCE: Arenaviruses are important viral pathogens that cause common and in some case severe infections in humans. Among them, Hemorrhagic Fever (HF) arenaviruses represent a serious public health problem as well as a threat as agents of bioterrorism. No licensed anti-arenavirus vaccines are available, and current anti-arenavirus therapy is limited to the use of ribavirin, which is only partially effective and often associated with severe side effects. Our goal is to generate a valid surrogate system that facilitates the study of HF New World members in the family under less-strict biosafety conditions by generating a single-cycle infectious, reporter-expressing, attenuated vaccine strain Candid#1.

描述（由申请人提供）：体育症病毒包括与高发病率和显着死亡率有关的人类出血热（HF）疾病的几种病因。此外，作为生物恐怖主义的特工，武器化的体育症病毒形式构成了严重的威胁。缺乏许可的疫苗和当前的抗嗜动病毒疗法受到了竞技病毒的严重关注，而限量使用了非标签的利巴韦林，这仅是部分有效的，并且通常与严重的副作用有关。 HF竞技场病毒研究的局限性包括在生物安全水平（BSL）4实验室中操纵这些药物的现场形式，以及对检测病毒的次要测定的要求。开发有效的单周期感染性代孕剂，这些代孕剂允许在较少破坏的BSL2设施下研究HF Arenavirus，并表达记者基因，以方便病毒检测，将促进使用安全，敏感和特定的筛选鉴定与安全，敏感和特定的筛选分析与高渗透率筛查（HTS）技术的鉴定。此外，它们还将代表有前途的疫苗方法，这些方法避免了对可能导致疾病的疫苗菌株的潜在失控复制的关注。最近，我们首次描述了单周感染性，表达记者的，旧世界淋巴细胞性粘膜炎病毒病毒（LCMV），在那里我们用记者的绿色荧光蛋白（GFP），RLCMV？gp/gp/gp/gp/gpp。通过与细胞系组成表达LCMV GP的遗传转移来实现传染病。该系统使我们能够研究病毒的多个方面，并开发筛查测定法，以检测和定量中和抗体和病毒抑制剂的鉴定。我们还能够通过产生表达LASV GP的细胞系和LASV GP pseudotypyed rlcmv？gp/gfp，将工作扩展到旧世界体育病毒的HF成员LASSA病毒（LASV）。由于旧世界和新世界竞技场病毒的储层，用于病毒的细胞受体使用以及RNA基因组组成的使用有所不同，因此必须开发有效的替代物来研究新世界Arenavirus，这是研究人员为研究人员提供一种研究病毒生物学，识别抗病毒剂的所有方面的方式 在不太严格的生物安全实验室下，针对新世界体育症病毒开发疫苗。在此应用程序中，我们建议通过产生单周感染性，表达记者，减毒的疫苗菌株候选＃1（RCAN？GP/GFP），将最近描述的技术扩展到新世界体育馆的研究。 公共卫生相关性：体育症病毒是引起常见的重要病毒病原体，在某些情况下是人类的严重感染。其中，出血热（HF）体育症病毒代表了一个严重的公共卫生问题，也是生物恐怖剂的威胁。没有许可的抗嗜动病毒疫苗可用，并且当前的抗动脉粥样硬病毒疗法仅限于使用利巴韦林，这仅是部分有效的，并且通常与严重的副作用有关。我们的目标是生成有效的替代系统，该系统通过产生单周感染性，表达记者，减弱的疫苗菌株念珠菌的单周感染性，表达疫苗的念珠菌，从而促进家庭中HF新世界成员的研究。