Development of Novel Pseudoinfectious Flavivirus Vaccines

新型伪传染性黄病毒疫苗的研制

• 批准号: 7649094
• 负责人: PETER W. MASON
• 金额: $ 32.47万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649094
• 关键词: Address; Alphavirus; Animal Diseases; Animals; Antigens; Attenuated Live Virus Vaccine; Binding; Capsid; Categories; Cell Line; Cells; Cellular Immunity; Chimera organism; Dengue; Dengue Virus; Development; Disease; E protein; Encephalitis; Engineering; Family; Flavivirus; Funding; Genes; Genome; Genome Components; Goals; Growth; Immune response; Immunity; In Vitro; Infection; Inorganic Sulfates; Japanese encephalitis virus; Laboratories; Langat virus; Life; Lymphoid Tissue; Methods; Modeling; Morbidity - disease rate; Mus; Names; Normal Cell; Omsk Hemorrhagic Fever; PTPN11 gene; Point Mutation; Primates; Production; Property; Proteins; Recording of previous events; Replicon; Side; Structural Protein; Structure; System; Technology; Testing; Tick-Borne Encephalitis; Unspecified or Sulfate Ion Sulfates; Vaccinated; Vaccines; Vertebral column; Viral; Viral Proteins; Virulent; Virus; Virus Diseases; Virus Replication; West Nile virus; Yellow fever virus; base; biodefense; cell type; citrate carrier; env Gene Products; immunogenicity; in vivo; man; novel; particle; pathogen; prevent; stable cell line; vaccine development; vector

Viruses in the flavivirus family populate all three of NIAID's Category A, B, and C lists of viruses. Vaccines are needed for multiple flavivirus diseases. To address this need, we have engineered a capsid (C) gene-deleted flavivirus vaccine platform that we have named RepliVAX. RepliVAX can infect cells in vitro and in vivo, but cannot produce live virus. However, RepliVAX-infected cells produce a key viral immunogen, a sub-viral particle (SVP) that contains the flavivirus envelope proteins, prM/M and E. SVPs have known ability to provide flavivirus immunity in man. RepliVAX can be propagated using two different systems. The first method is based on growth in Cexpressing cell lines that produce large amounts of C protein from a non-cytopathic alphavirus vector. In this system, RepliVAX is grown in a format similar to that used for production of classical live attenuated vaccines. The second system for RepliVAX production involves the co-cultivation of RepliVAX with helper particles that encode a flavivirus genome expressing a functional form of C, but deleted for the prM and E genes. This second system, referred to throughout this proposal as the "two-component" genome system has the advantage that it permits RepliVAX production on any type of cell line. RepliVAX vaccines based on the genomes of yellow fever virus (YFV) and West Nile virus (WNV) can protect animals from YF and WN encephalitis. We have also applied the widely used "chimeric" flavivirus technology to generate a WNV-based RepliVAX encoding the prM/E structural proteins of Japanese encephalitis virus (JEV). This RepliVAX JE protects animals from JE, suggesting that using the highly efficient WNV replication "engine" to drive chimeric RepliVAX provides a breakthrough for flavivirus vaccine development. The goal of this 6-year project will be the development of RepliVAX as a platform, by extending it to the production of safe and effective vaccines for the most widely circulating emerging flavivirus disease (dengue) and two biodefense threat diseases with known high morbidity and a history of weaponization {Omsk hemorrhagic fever (OHF) and tick-borne encephalitis (TBE)}. To develop RepliVAX into a universal flavivirus vaccine platform, we have devised the following specific aims. 1: Determine if stable cells or the "two-component" system is best for RepliVAX production. 2. Determine which RepliVAX backbone is superior for producing effective TBE vaccines. 3: Generate RepliVAX chimeras with dengue virus (DENV) that grow to high titers. 4: Create chimeric RepliVAX expressing the prM/E and NS1 proteins and determine if these are superior to prM/E. 5. Determine if RepliVAX can be retargeted to enhance its efficacy.

黄病毒家族中的病毒存在于NIAID的所有A、B和C类病毒列表中。 多种黄病毒疾病需要疫苗。为了满足这一需求，我们设计了一个衣壳 (C)基因缺失黄病毒疫苗平台，我们将其命名为RepliVAX。RepliVAX可以感染 体外和体内，但不能产生活病毒。然而，RepliVAX感染的细胞产生一种关键的病毒 一种亚病毒颗粒(SVP)，包含黄病毒囊膜蛋白PRM/M和E.SVPS 已知有能力为人类提供黄病毒免疫力。 RepliVAX可以使用两个不同的系统进行传播。第一种方法是基于C++的Growth 从非细胞病态甲型病毒载体产生大量C蛋白的细胞系。在……里面 该系统RepliVAX的生长格式类似于用于生产经典实况衰减的格式 疫苗。RepliVAX生产的第二个系统涉及与Helper共培养RepliVAX 编码黄病毒基因组的颗粒，表达功能性形式的C，但缺失PRM和E 基因。这第二个系统，在整个提案中被称为“双组分”基因组系统 它的优点是允许在任何类型的细胞系上生产RepliVAX。 基于黄热病病毒和西尼罗河病毒基因组的RepliVAX疫苗可以 保护动物免受YF和WN脑炎的侵袭。我们还应用了广泛使用的“嵌合”黄病毒。 编码日本株PRM/E结构蛋白的西尼罗河病毒RepliVAX的构建技术 脑炎病毒(JEV)这种RepliVAX JE保护动物免受乙脑的侵袭，这表明使用高度 高效WNV复制引擎驱动嵌合RepliVAX为黄病毒疫苗提供突破 发展。这个为期6年的项目的目标将是开发RepliVAX作为一个平台，通过 将其扩展到生产安全有效的疫苗，用于最广泛传播的新兴疫苗 黄病毒病(登革热)和两种生物防御威胁疾病，已知发病率高，有 武器化{鄂木斯克出血热和壁虱传播的脑炎}。 为了将RepliVAX开发成一个通用的黄病毒疫苗平台，我们设计了以下具体方案 目标。1：确定稳定细胞或“双组分”系统哪个最适合RepliVAX的生产。2. 确定哪种RepliVAX主干更适合生产有效的TBE疫苗。3：生成 带有登革病毒(DENV)的RepliVAX嵌合体，这些嵌合体生长到高滴度。4：创建嵌合RepliVAX 表达PRM/E和NS1蛋白，并确定它们是否优于PRM/E。 RepliVAX可以被重定向以增强其疗效。