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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 包括委内瑞拉（VEEV），东部（EEEV）和西马脑炎病毒（WEEV）在内的α病毒是生物战和恐怖主义（BWT）的高度发达的药物，并且自然而然地出现了人畜共患病毒。目前，针对这些病毒的生物陷入困境不足，在平民或军事人员暴露时可能会造成广泛的发病率和死亡率。有效的，有执照的疫苗，关键的第一线防御和生物化研究的重要工具非常需要。我们专注于活病毒疫苗，以利用其高水平的抗原表现，诱导体液和细胞介导的免疫的能力，以及更快，更长的寿命抗体水平的特征是暴露于复制病毒的特征。在提议延续我们的α病毒疫苗开发项目中，我们将完善并进一步验证一组候选疫苗的安全性和功效，以防止VEEV，EEEV和WEEV。将在嵌合疫苗基因组中测试其他衰减策略，以利用我们最近确定某些α病毒的衣壳蛋白抑制宿主细胞转录，这是毒力的重要机制。作为进一步的安全措施，将开发出一种新的遗传策略，以使嵌合疫苗病毒在体外和体内复制的能力完全衰弱。最后，为了进一步评估这些嵌合体对人类疫苗接种的潜力，在对安全性，免疫原性和衰减的初步评估中，将使用2种不同的sindbis/sindbis/东马脑炎疫苗接种疫苗进行免疫。除了保护平民和军事人口免受预期的α病毒BWT事件，以及对从事关键BWT和公共卫生研究的实验室人员的保护外，我们还可以利用我们开发的方法，以快速有效地开发新的疫苗，以针对新认识的，新兴的Alphaviruess或一种工程化的Alphavirus武器开发新的疫苗。它们还将有助于保护新世界许多地区自然接触这些人畜共患药物的风险。 到目前为止，我们已经使用Cynomolgus Macaques（Macaca fascicularis）完成了一项使用候选EEEV疫苗的疗效研究。 用北美（Sind-Naeeevv）或南美（Sind-Saeeevv）变种疫苗的sindbis病毒（Sind-Naeeevv）或盐水免疫的假疫苗。 确定动物的叶片并确定抗体产生（α-EEEV IgG）。 这些动物在靶标致命剂量（1.0E+07 PFU吸入）处被气溶胶北美EEE病毒（FL-9393）挑战。 接受SIND-NAEEEVV变体的动物大多受到保护（5/6生存； 83％），而南美变种（Sind-Saeeevv）接种疫苗的动物或Sham-vaccined（盐水）因暴露而死亡（1/6; 1/6; 16％; 16％; 16％和0/4; 0/4; 0％; 0％; 0％;分别生存。 对动物挑战后对动物的远程监测表明，动物成功接种了疫苗，核心温度正常变化或心率和心电图异常的正常昼夜变化没有中断。 替代地，受影响的动物在核心温度和ECG信号中的破坏中经历了重大变化（高温）。 组织（特别是大脑）的组织病理学分析表明病毒浸润和明显的脑炎。 我们正在重复Cynomolgus猕猴中的该实验，以确认SIND-EEEVV在此模型中的功效。