Development of a Bacteriophage Vaccine Vector System

噬菌体疫苗载体系统的开发

• 批准号: 7781345
• 负责人: Steve Bende
• 金额: $ 17.48万
• 依托单位: BACILLIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-11 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7781345
• 关键词: Address; Animals; Antibody Formation; Antigens; Avian Influenza; Awareness; Bacteria; Bacteriophages; Biological; Biological Assay; Biological Products; Bioterrorism; Capsid; Cell Culture Techniques; Cells; Cellular Immunity; Code; Computer Systems Development; Cystovirus; Data; Development; Double-Stranded RNA; Elements; Emergency Situation; Ensure; Escherichia coli; Event; Generations; Genes; Genetic Transcription; Genomic Segment; Goals; Human; Immune; In Vitro; Infection; Influenza; Influenza A Virus, H5N1 Subtype; Influenza A virus; Intramuscular; Laboratory Animals; Lipids; Lung; Mammalian Cell; Measurable; Measures; Membrane; Mus; Nucleic Acid Vaccines; Nucleotides; Preparation; Principal Investigator; Procedures; Process; Production; Proteins; Protocols documentation; Public Health; RNA; RNA Phages; RNA replication; RNA-Directed RNA Polymerase; Readiness; Recombinants; Regimen; Reoviridae; Reporter; Research; Risk; Route; Safety; Scientist; Serotyping; Splenocyte; Subunit Vaccines; System; Testing; TimeLine; Transcription Initiation; Translations; Vaccinated; Vaccine Antigen; Vaccines; Virus; Virus-like particle; base; biodefense; cytokine; experience; flexibility; flu; immunogenicity; in vivo; influenza virus vaccine; influenzavirus; novel; pandemic disease; pandemic influenza; pathogen; prevent; protein expression; public health relevance; response; seasonal influenza; success; synthetic construct; tool; vector; vector vaccine; weapons

DESCRIPTION (provided by applicant): The goal of the studies in this proposal is to further develop a process that enables rapid production, purification and application of a bacteriophage vector system (BVS) platform expressing influenza A virus-like particles (Flu-VLPs). This goal is warranted as BVS is a unique vector that can be rapidly produced in bacteria, affords the safety profile associated with RNA-based vectors and has the potential for dual application as a transient expression system to produce subunit vaccines in vitro and as a vaccine vector directly administrable to humans or animals in vivo. BVS is derived from the double- stranded RNA (dsRNA) Cystovirus bacteriophage phi-8. To render this bacteriophage for vaccine applications the PI and coworkers have genetically altered genomic segment-S (gsS) and genomic segment-M (gsM) of phi-8, and launched the BVS recombinant phi-8 capsids in Escherichia coli host strains. In preliminary studies purified BVS capsids have been shown to express reporter proteins and vaccine antigens in mammalian cells in vitro. This background experience and evidence supports the proposal to further develop BVS capsids as a platform for emergency preparedness/biodefense applications. The PI provides justification that successful development of this system will create a public health tool and biodefense countermeasure, which under normal circumstances is capable of producing conventional subunit vaccines; yet, it is proposed that BVS capsids can be deployed as a vaccine vector from sequence identification through to vaccine release in under a month during emergency events where such a risk may be warranted. In this manner, this system integrates the advantages of manufacturing in bacteria and the utility and safety of non-replicating RNA-based nucleic acid vaccine vectors with the need for rapid response timelines to ensure broad emergency preparedness against emerging influenza virus serotypes such as H5N1. The central hypothesis that will be tested in the proposed studies below is that BVS capsids have the potential to serve as both a vaccine producer and vaccine vector. To address this hypothesis the PI proposes to (i) To develop a procedure to rapidly produce purified and bioactive BVS capsids that express Flu-VLPs and (ii) To characterize the immunogenicity of Flu-VLPs and BVS capsids expressing Flu-VLPs in laboratory animals. It is envisaged that successful completion of these aims will create an effective BVS capsid and Flu-VLP production system that can be adapted for manufacturing of both seasonal influenza and emerging influenza subtypes, and deployed significantly more rapidly than other vaccine modes. In addition, the proposed studies will provide fundamental information germane to the adaptation of this system to become an operational platform for rapid development and deployment of public health and biodefense vaccines. PUBLIC HEALTH RELEVANCE: A renewed awareness of the urgent need to devise strategies for the rapid development, manufacture and distribution of vaccines was invoked by the threat of avian influenza subtype H5N1 to spawn a catastrophic human pandemic and the increased threat of bioterrorism with other biological agents. The goal of the studies in this proposal is to provide a manufacturing platform that enables rapid production, purification and administration of vaccines. Furthermore, the studies herein will generate vaccine components and manufacturing procedures that will have an immediate utility in the control of both seasonal and pandemic influenza. Additionally, it is envisaged that this system will serve as an operational platform for rapid development and deployment of a broad array of public health and biodefense vaccines.

描述（由申请人提供）：本提案中的研究的目的是进一步开发一个过程，该过程能够快速生产，纯化和应用噬菌体矢量系统（BVS）平台，表达流感的一种类似病毒的颗粒（FLU-VLPS） 。该目标是必要的，因为BVS是一个独特的向量，可以在细菌中迅速产生，提供与基于RNA的矢量相关的安全性，并且有可能在体外产生亚基疫苗的瞬时表达系统，并且作为一种疫苗。载体直接适用于人类或动物的体内。 BVS源自双链RNA（DSRNA）膀胱病毒噬菌体PHI-8。为了使这种噬菌体用于疫苗应用，PI和同事在PHI-8的基因组段S（GSS）和基因组段M（GSM）中具有遗传变化，并在Escherichia coli宿主株中启动了BVS重组PHI-8 Capsids。在初步研究中，纯化的BVS衣壳已被证明可以在体外表达哺乳动物细胞中的报告蛋白和疫苗抗原。这种背景经验和证据支持进一步开发BVS Capsids作为紧急准备/Biodefense应用程序的平台的提议。 PI提供了合理的理由，即该系统的成功开发将创建公共卫生工具和生物化的对策，在正常情况下，该工具能够生产常规的亚基疫苗；然而，有人提出，在紧急事件中，可以将BVS CAPSID部署为从序列识别到疫苗释放的疫苗向量，从而在紧急事件中释放疫苗。通过这种方式，该系统整合了细菌中制造的优势以及非复制RNA的基于RNA的核酸疫苗载体的效用和安全性，需要快速响应时间表，以确保针对新兴的流感病毒血清型（例如H5N1）进行广泛的紧急准备。下面提出的研究将在下面提出的研究中进行检验的中心假设是，BVS衣壳具有作为疫苗生产者和疫苗载体的潜力。为了解决这一假设，PI提议（i）开发一种程序，以快速产生表达流感VLP的纯净和生物活性的BVS衣壳，以及（ii）表征流感VLPS和BVS Capsids在实验室动物中表达流感vlps的免疫原性。可以预见的是，这些目标的成功完成将创建有效的BVS Capsid和Flu-VLP生产系统，可以适应以制造季节性流感和新兴的流感亚型，并且比其他疫苗模式更快地部署了更快的速度。此外，拟议的研究将为适应该系统的适应提供基本信息，以成为快速开发和部署公共卫生和生物污染疫苗的运营平台。公共卫生相关性：对迫切需要为快速开发，疫苗的生产和分配制定策略的重新认识，这是由于鸟类流感亚型H5N1的威胁所援引的，以产生灾难性的人类大流行病，并增加生物恐怖主义的威胁与其他生物恐怖主义的威胁。该提案中研究的目的是提供一个制造平台，以促进疫苗的快速生产，纯化和给药。此外，本文的研究将产生疫苗成分和制造程序，这些程序将立即控制季节性和大流行流感。此外，可以预见，该系统将成为快速开发和部署各种各样的公共卫生和生物污染疫苗的运营平台。