Nodavirus-based RNA replicon vaccines for tick-borne encephalitis virus

基于诺达病毒的 RNA 复制子疫苗，用于蜱传脑炎病毒

• 批准号: 7649742
• 负责人: KYLE L JOHNSON
• 金额: $ 16.46万
• 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/7649742
• 关键词: Animals; Antibodies; Arthropods; Asia; Bioterrorism; Bos taurus structural-GP protein; Categories; Cells; Central European Encephalitis; Development; Encephalitis; Encephalitis Viruses; Europe; Goals; Human; Immune response; Inactivated Vaccines; Individual; K-Series Research Career Programs; Lead; Mammalian Cell; Mediating; Messenger RNA; Mus; National Institute of Allergy and Infectious Disease; Neurologic; Nonstructural Protein; North America; PTPN11 gene; Powassan virus; Proteins; Public Health; RNA; RNA Interference; RNA replication; Replicon; Reporting; Research; Risk; Russian Spring-Summer Encephalitis; Saccharomyces cerevisiae; Survivors; Testing; Tick-Borne Encephalitis; Tick-Borne Encephalitis Virus; Tick-Borne Encephalitis Viruses; Ticks; United States; Vaccines; Viral Proteins; Virus; Yeasts; aerosolized; base; biodefense; cell mediated immune response; concept; immunogenicity; mortality; novel; pathogen; prevent; protein expression; replicon vaccine; transmission process; vaccine development; viral RNA

Tick-borne encephalitis (TBE) viruses are important human pathogens, particularly in Europe and Asia; one TBE virus, Powassan virus, is endemic to North America. TBE viruses, which are transmitted by ticks, can cause fatal encephalitis in humans. Russian spring-summer encephalitis virus and Central European Encephalitis virus result in mortality of 20% and 1-2%, respectively. In each case, survivors are at risk for developing neurological sequelae. An inactivated virus vaccine protects individuals exposed to ticks in Europe but this vaccine has not been approved for use in the United States. Despite the existence of the vaccine, several thousand cases of TBE encephalitis are reported each year. The National Institute for Allergy and Infectious Disease (NIAID) Biodefense Research Agenda identifies TBE as a Category C priority pathogen on the basis of its transmission by an arthropod host, its infectivity on aerosolized exposure, and its potential threat to public health. Also, of particular concern is the possibility that TBE viruses might be used as agents of bioterrorism. Therefore, development of a vaccine against TBEV consistent with the overall goals of the WRCE as well as a national biodefense priority. The goals of this Career Development Award proposal are to develop a candidate vaccine for tick-borne encephalitis virus and to test its immunogenicity in mice. This proposal incorporates three novel concepts, namely the use of Nodamura virus (NoV) RNA replicons to amplify TBEV mRNAs in the yeast Saccharomyces cerevisiae, the inclusion of an NoV protein that suppresses host cellular defenses like RNA interference, and inoculation of animals with purified total yeast RNA containing amplified NoV-TBEV RNAs. The specific aims of this proposal are to: 1. Construct NoV RNA2-based replicons that contain TBEV structural (glycoprotein E; M) or nonstructural (NS1) proteins and determine the extent to which RNA replication amplifies NoV2-gpE, NoV-M and NoVNS1 mRNA and protein levels in yeast and mammalian cells. 2. Determine the extent to which suppression of RNA interference by an NoV nonstructural protein enhances the efficiency of protein expression from yeast-derived NoV RNA replicons. 3. Evaluate the TBEV structural (gpE, M) 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. Tick-borne encephalitis (TBE) viruses are serious human pathogens that pose a threat to our public health and our national biosafety. Our goal is to develop safe and effective vaccine candidates that will prevent the spread of TBE.

tick传播的脑炎（TBE）病毒是重要的人类病原体，尤其是在欧洲和亚洲；一 Powassan病毒TBE病毒是北美特有的。由tick传输的病毒可以 引起人类致命性脑炎。俄罗斯春季夏末病毒和中欧 脑炎病毒的死亡率分别为20％和1-2％。在每种情况下，幸存者都有 发展神经后遗症。灭活的病毒疫苗可保护暴露于tick虫的个体 欧洲，但这种疫苗尚未批准在美国使用。尽管存在 疫苗，每年有数千例TBE脑炎。国家研究所 过敏和传染病（NIAID）生物陷入困境研究议程将TBE视为c优先级 病原体是根据节肢动物宿主传播的，其对雾化暴露的感染力，以及 它对公共卫生的潜在威胁。另外，特别关心的是病毒可能是 用作生物恐怖主义的药物。因此，开发针对TBEV的疫苗与 WRCE的总体目标以及国家生物陷入困境的优先事项。这个职业发展的目标 奖励提案是为tick传播脑炎病毒开发候选疫苗，并测试其 小鼠的免疫原性。该提案包含了三个新颖概念，即使用Nodamura病毒 （nov）RNA复制子在酵母菌中扩增TBEV mRNA，酿酒酵母，包括 抑制RNA干扰等宿主细胞防御的Nov蛋白质，并接种具有 纯化的含有扩增的Nov-TBEV RNA的酵母RNA。该提案的具体目的是： 1。构造基于Nov RNA2的复制子，其中包含TBEV结构（糖蛋白E; M）或非结构性 （NS1）蛋白质并确定RNA复制在多大程度上放大Nov2-GPE，Nov-M和Novns1 酵母和哺乳动物细胞中的mRNA和蛋白质水平。 2。确定NOV非结构蛋白对RNA干扰的抑制程度增强 酵母衍生的Nov RNA复制子的蛋白质表达效率。 3。评估TBEV结构（GPE，M）和非结构（NS1）蛋白作为潜在的候选疫苗 通过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。 tick传播脑炎（TBE）病毒是姿势的严重人类病原体 对我们的公共卫生和国家生物安全构成威胁。我们的目标是开发安全有效的疫苗 可以防止TBE传播的候选人。