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.

森林脑炎病毒是重要的人类病原体，特别是在欧洲和亚洲； TBE病毒，鲍瓦桑病毒，是北美特有的病毒。由扁虱传播的TBE病毒可以 在人类身上引起致命的脑炎。俄罗斯春夏脑炎病毒与中欧 脑炎病毒导致的死亡率分别为20%和1-2%。在每一种情况下，幸存者都面临着 发展成神经后遗症。灭活病毒疫苗保护暴露在扁虱中的个人 但这种疫苗尚未被批准在美国使用。尽管存在 在接种疫苗后，每年都有数千例脑炎病例报告。美国国立卫生研究院 变态反应和传染病(NIAID)生物防御研究议程将TBE确定为C类优先事项 病原体根据其由节肢动物宿主传播、其在气雾化暴露中的传染性以及 它对公众健康的潜在威胁。此外，特别令人担忧的是，TBE病毒可能是 被用作生物恐怖主义的代理人。因此，开发一种与TBEV一致的疫苗 WRCE的总体目标以及国家生物防御优先事项。这份职业发展的目标 获奖建议是开发森林脑炎病毒的候选疫苗，并测试其 小鼠的免疫原性。这项提议包含了三个新概念，即使用新村病毒 (Nov)在酿酒酵母中扩增TBEV mRNAs的RNA复制子，包括 抑制宿主细胞防御的NOV蛋白，如RNA干扰和动物接种 纯化的酵母总RNA含有扩增的NOV-TBEV RNA。这项建议的具体目的是： 1.构建含TBEV结构蛋白(糖蛋白E；M)或非结构蛋白的基于nov RNA2的复制体 (NS1)蛋白质，并确定RNA复制放大NoV2-GPE、Nov-M和NoVNS1的程度 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 酵母来源的nov RNA复制子的蛋白质表达效率。 3.评估TBEV结构蛋白(GPE，M)和非结构蛋白(NS1)是否为潜在的候选疫苗 通过定义接种小鼠对这些蛋白质的体液和细胞介导的免疫反应。 与公共卫生的相关性。森林脑炎(TBE)病毒是一种严重的人类病原体 对我们的公共健康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 将防止TBE传播的候选人。