VACCINE DEVELOPMENT FOR ALPHAVIRUSES

甲病毒疫苗的开发

• 批准号: 8173021
• 负责人: CHAD J. ROY
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173021
• 关键词: Aerosols; Affect; Alphavirus; American; Animals; Antibodies; Antibody Formation; Antigen Presentation; Biological Warfare; Brain; Breathing; Capsid Proteins; Cells; Cellular Immunity; Characteristics; Chimera organism; Circadian Rhythms; Computer Retrieval of Information on Scientific Projects Database; Culicidae; Dose; Eastern Equine Encephalitis Virus; Eastern Equine Encephalomyelitis; Electrocardiogram; Encephalitis; Engineering; Event; Exposure to; Fever; Funding; Genetic; Genetic Transcription; Genome; Grant; Heart Rate; Hemorrhage; Human; Immunoglobulin G; In Vitro; Institution; Laboratory Personnel; Licensing; Life; Macaca; Macaca fascicularis; Measures; Military Personnel; Modeling; Monitor; Morbidity - disease rate; Population; Populations at Risk; Research; Research Methodology; Research Personnel; Resources; Safety; Saline; Signal Transduction; Sindbis Virus; Source; South American; Temperature; Terrorism; Testing; Tissues; United States National Institutes of Health; Vaccinated; Vaccination; Vaccines; Variant; Viral; Virulence; Virus; Western Equine Encephalitis Virus; attenuation; biodefense; experience; immunogenicity; in vivo; mortality; novel; novel vaccines; public health research; research study; tool; vaccine candidate; vaccine development; weapons

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Alphaviruses including Venezuelan (VEEV), eastern (EEEV) and western equine encephalitis viruses (WEEV) are highly developed agents of biological warfare and terrorism (BWT) and important, naturally emerging zoonotic viruses. Current biodefense against these viruses is inadequate, and widespread morbidity and mortality could be inflicted upon exposure of civilians or military personnel. Effective, licensed vaccines, critical first lines of defense and important tools for biodefense research, are badly needed. We have focused on live virus vaccines to take advantage of their high levels of antigen presentation, their ability to induce both humoral and cell-mediated immunity, and the faster and longer lived antibody levels characteristic of exposure to replicating viruses. In the proposed continuation of our alphavirus vaccine development project, we will refine and further validate the safety and efficacy of a set of vaccine candidates against VEEV, EEEV and WEEV. Additional attenuation strategies will be tested in the chimeric vaccine genomes to take advantage of our recent determination that the capsid proteins of some alphaviruses inhibit host cell transcription, an important mechanism of virulence. As a further safety measure, a novel genetic strategy will be developed to completely debilitate the ability of the chimeric vaccine viruses to replicate in mosquito cells, both in vitro and in vivo. Finally, to further evaluate the potential of these chimeras for vaccination of humans, Cynomolgus macaques will be immunized with 2 different Sindbis/Eastern equine encephalitis vaccine candidates in a preliminary assessment of safety, immunogenicity and attenuation. In addition to protection of civilian and military populations from an anticipated alphavirus BWT event, and protection of laboratory personnel doing critical BWT and public health research, the methods we will develop can be exploited to rapidly and efficiently develop new vaccines against newly recognized, emerging alphaviruses or an engineered alphavirus weapon. They will also be useful for protecting populations at risk of natural exposure to these zoonotic agents in many parts in the New World. Thus far, we have completed an efficacy study using the candidate EEEv vaccines using cynomolgus macaques (Macaca fascicularis). Animals were primed with a Sindbis virus vectored North American (SIND-NAEEEvv) or South American (SIND-SAEEEvv) variant vaccine or sham immunized with saline. Animals were bled and antibody production (alpha-EEEv IgG) was determined. The animals were challenged with a North American origin EEE virus (FL-9393) by aerosol at a target lethal dose (1.0E+07 PFU inhaled). Animals receiving SIND-NAEEEvv variant were mostly protected (5/6 survival; 83%) whereas animals vaccinated with the South American variant (SIND-SAEEEvv) or sham-vaccinated (saline) died as a result of exposure (1/6; 16% and 0/4; 0% survival respectively). Telemetric monitoring of the animals post challenge showed the animals successfully vaccinated experienced minimal to no disruption in normal diurnal variation in core temperature or abnormalities in heart rate and ECG. The affected animals, alternatively, experienced significant changes (hyperthermia) in core temperature and disruption in ECG signal. Histopathological analysis of tissues, specifically the brain, indicated viral invasion and significant encephalitis. We are repeating this experiment in cynomolgus macaques to confirm the efficacy of SIND-EEEvv in this model.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 包括委内瑞拉（VEEV）、东方（EEEV）和西方马脑炎病毒（WEEV）的甲病毒是高度发达的生物战和恐怖主义（BWT）因子和重要的自然出现的人畜共患病毒。目前对这些病毒的生物防御是不够的，平民或军事人员接触这些病毒可能会造成广泛的发病率和死亡率。有效的、有许可证的疫苗，关键的第一道防线和生物防御研究的重要工具，是迫切需要的。我们专注于活病毒疫苗，以利用其高水平的抗原呈递，诱导体液和细胞介导的免疫的能力，以及暴露于复制病毒的更快和更长寿命的抗体水平。在我们甲病毒疫苗开发项目的拟议延续中，我们将完善并进一步验证一组针对VEEV、EEEV和WEEV的候选疫苗的安全性和有效性。将在嵌合疫苗基因组中测试其他减毒策略，以利用我们最近确定的一些甲病毒的衣壳蛋白抑制宿主细胞转录（毒力的重要机制）的优势。作为进一步的安全措施，将开发一种新的遗传策略，以完全削弱嵌合疫苗病毒在体外和体内蚊子细胞中复制的能力。最后，为了进一步评价这些嵌合体用于人类疫苗接种的潜力，在安全性、免疫原性和减毒的初步评估中，将用2种不同的辛德毕斯/东部马脑炎候选疫苗免疫食蟹猴。除了保护平民和军事人群免受预期甲病毒BWT事件的影响，以及保护进行关键BWT和公共卫生研究的实验室人员外，我们将开发的方法可以用于快速有效地开发针对新识别的新甲病毒或工程甲病毒武器的新疫苗。它们还将有助于保护新世界许多地区面临自然暴露于这些人畜共患病病原体风险的人群。 到目前为止，我们已经使用食蟹猴（Macaca fascicularis）完成了候选EEEv疫苗的有效性研究。 动物用辛德毕斯病毒载体的北美（SIND-NAEEEvv）或南美（SIND-SAEEEvv）变体疫苗致敏或用盐水假免疫。 对动物流血的并测定抗体产生（a-EEEv IgG）。 以目标致死剂量（吸入1.0E+07 PFU）通过气溶胶用北美来源的HIV病毒（FL-9393）对动物进行攻毒。 接受SIND-NAEEEvv变体的动物大多数受到保护（5/6存活; 83%），而接种南美变体（SIND-SAEEEvv）或假接种（生理盐水）的动物因暴露而死亡（分别为16; 16%和0/4; 0%存活）。 攻毒后动物的遥测监测显示，成功接种疫苗的动物在核心温度的正常昼夜变化或心率和ECG异常方面发生了极轻微至无干扰。 受影响的动物，或者，经历了显着的变化（体温过高），在核心温度和ECG信号中断。 组织的组织学分析，特别是大脑，表明病毒入侵和严重的脑炎。 我们在食蟹猴中重复该实验，以确认SIND-EEEvv在该模型中的疗效。