Nodavirus-Based Vaccines for West Nile Virus (pilot)

基于诺达病毒的西尼罗河病毒疫苗（试点）

• 批准号: 7858092
• 负责人: KYLE L JOHNSON
• 金额: $ 7.19万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7858092
• 关键词: Animals; Antibodies; Arthropods; Bioterrorism; Bos taurus structural-GP protein; Categories; Cells; Development; Disease; Goals; Hepatitis B Vaccines; Human; Human Virus; Immune response; Insecta; Lead; Mammalian Cell; Mammals; Mediating; Messenger RNA; Mission; Mus; National Institute of Allergy and Infectious Disease; PTPN11 gene; Pathogenicity; Plants; Proteins; Public Health; RNA; RNA replication; Replicon; Research; Research Project Grants; Saccharomyces cerevisiae; Source; System; Testing; Texas; Transfection; United States; United States National Institutes of Health; Universities; Vaccines; Virus; West Nile virus; Yeasts; aerosolized; base; biodefense; cell mediated immune response; immunogenicity; mortality; novel; pathogen; prevent; programs; public health priorities; transmission process; vaccine candidate; viral RNA

West Nile virus (WNV) is a serious human pathogen with a mortality rate of 0.1%. The National Institute for Allergy and Infectious Disease (NIAID) Biodefense Research Agenda classifies WNV as a Category B Priority Pathogen on the basis of its transmission by an arthropod host, its infectivity on aerosolized exposure, and its potential threat to public health. Of particular concern is the possibility that West Nile virus might be used as an agent of bioterrorism. There is currently no vaccine for WNV available for use in humans. Thus, development of a WNV vaccine is a national biodefense and public health priority that is consistent with the overall goals of NIH and MBRS. The goals of this Pilot Research Project are to develop a candidate vaccine for West Nile virus and to test its immunogenicity in mice. This proposal incorporates two novel concepts: the use of Nodamura virus (NoV) RNA replicons to amplify WNV mRNAs in the yeast Saccharomyces cerevisiae and inoculation of animals with purified total yeast RNA containing amplified NoV-WNV RNAs. The advantages of the nodavirus expression system as a source of vaccine material include exponential amplification of the chimeric mRNA; its ability to initiate RNA replication in a wide variety of host cells, including those from mammals, insects, plants, and yeast, when the RNA is introduced by transfection; and the lack of pathogenicity of NoV for humans. Yeast cells provide a safe and inexpensive source of vaccine material, and are currently used to produce Hepatitis B vaccines. The specific aims of this proposal are: 1. Construct NoV RNA2-based replicons that contain WNV structural (glycoprotein E, gpE) or nonstructural (NS1) proteins and determine the extent to which RNA replication amplifies NoV2-gpE and NoV-NS1 mRNA and protein levels in yeast and mammalian cells 2. Evaluate the WNV structural (gpE) and nonstructural (NS1) proteins as potential vaccine candidates by defining the humoral and cell-mediated immune responses to these proteins elicited in inoculated mice. Relevance to public health: West Nile virus is a severe human pathogen that poses a threat to the public health and biosafety of the United States. Our goal is to develop safe, effective candidate vaccines that will prevent the spread of West Nile virus-associated diseases.

西尼罗河病毒（WNV）是一种严重的人类病原体，死亡率为0.1％。国家研究所 过敏和传染病（NIAID）生物陷入困境研究议程将WNV归类为B类 优先病原体根据节肢动物宿主的传播，其感染性在雾化 暴露及其对公共卫生的潜在威胁。特别关心的是西尼罗河病毒的可能性 可以用作生物恐怖主义的代理。目前没有可用于WNV的疫苗可用于 人类。因此，WNV疫苗的开发是一种国家生物防御和公共卫生优先事项 与NIH和MBR的总体目标一致。该试点研究项目的目标是 开发用于西尼罗河病毒的候选疫苗，并在小鼠中测试其免疫原性。这个建议 结合了两个新颖的概念：使用Nodamura病毒（NOV）RNA复制子来扩增WNV mRNA 酿酒酵母的酵母菌和含有纯化的总酵母RNA的动物接种 放大了Nov-WNV RNA。诺达病毒表达系统的优势是疫苗的来源 材料包括嵌合mRNA的指数扩增；它在A中启动RNA复制的能力 RNA为 通过转染引入；以及人类缺乏致病性。酵母细胞提供安全的 疫苗材料的廉价来源，目前用于生产丙型肝炎疫苗。这 该提案的具体目的是： 1。构造基于Nov RNA2的复制子，其中包含WNV结构（糖蛋白E，GPE）或非结构性 （NS1）蛋白质并确定RNA复制放大Nov2-GPE和Nov-NS1的程度 酵母和哺乳动物细胞中的mRNA和蛋白质水平 2。将WNV结构（GPE）和非结构（NS1）蛋白评估为潜在的疫苗候选物 定义对接种小鼠引起的这些蛋白质的体液和细胞介导的免疫反应。 与公共卫生有关：西尼罗河病毒是一种严重的人类病原体，对公众构成威胁 美国的健康和生物安全。我们的目标是开发安全，有效的候选疫苗 防止西尼罗河病毒相关疾病的扩散。