Pathogenesis of Subtype IE Venezuelan Equine Encephalitis Virus

IE亚型委内瑞拉马脑炎病毒的发病机制

• 批准号: 8084210
• 负责人: Alison Paige Adams
• 金额: $ 11.89万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-15 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8084210
• 关键词: Address; Affect; Agriculture; Americas; Animals; Arboviruses; Area; Asses; Biological Assay; Biological Warfare; Bioterrorism; Blood; Categories; Cell Culture Techniques; Cells; Central America; Child; Chimera organism; Colombia; Commerce; Complement; Complex; Culicidae; Data; Dendritic Cells; Development; Disease; Disease Outbreaks; Doctor of Philosophy; Economics; Ecuador; Encephalitis; Environment; Epidemic; Equine mule; Equipment; Equus caballus; Faculty; Funding; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Determinism; Genome; Genomics; Genotype; Glycoproteins; Goals; Health; Human; Immune response; Immunologist; Immunology; Infection; Insecticides; Interferons; Intervention; Knowledge; Laboratories; Lead; Lesion; Leukocytes; Lymphoid; Measures; Mediating; Medical; Medicine; Mexico; Molecular; Mutation; National Institute of Allergy and Infectious Disease; Neurologic Symptoms; Nonstructural Protein; Pathogenesis; Pathogenicity; Phenotype; Population; Research; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Serotyping; Serum; Site; Structural Protein; Testing; Texas; Therapeutic Agents; Therapeutic Uses; Tissues; Training; Travel; Tropism; USSR; United States National Institutes of Health; Universities; Vaccines; Variant; Venezuela; Venezuelan Equine Encephalitis Virus; Venezuelan Equine Encephalomyelitis; Viral; Viral Genes; Viremia; Virulence; Virulent; Virus; Virus Diseases; base; cDNA Arrays; comparative; cytokine; design; disorder control; enzootic; epizootic; fetal; genetic technology; human disease; immunogenicity; immunoregulation; improved; in vivo; instructor; laboratory accident; macrophage; monocyte; mortality; neurovirulence; pathogen; peripheral blood; positional cloning; programs; public health relevance; recombinant virus; relating to nervous system; research study; skills; therapeutic vaccine; vaccine development; vector control

DESCRIPTION (provided by applicant): Candidate: Dr. Adams completed her DVM cum laude at Texas A&M University, and she completed her Ph.D. in immunology in Dr. Douglas Antczak's laboratory at Cornell University, studying mechanisms of fetal evasion from maternal immune responses. Dr. Adams will apply her skills as an immunologist to pursue her long-term goal of understanding the pathogenesis of alphaviral encephalitides, knowledge that will ultimately be used for development of effective vaccines/therapeutics. The proposed project is designed to bridge her transition from an Instructor to a permanent academic post in comparative medicine research. Research: Venezuelan equine encephalitis virus (VEEV) is a naturally emerging arbovirus that threatens human health throughout the Americas. Human epidemics only occur when there is efficient amplification of VEEV in equids. VEEV is also considered a potential bioterrorist agent, categorized by the NIAID as a Category B Priority Pathogen. A recent outbreak of an epizootic subtype IE VEEV underscores the continued threat of VEE emergence in human populations and what little we know about its ability to amplify in equids and alter virulence via mutations. The objective of this proposal is to dissect the role of structural and nonstructural proteins of epizootic subtype IE VEEV in causing efficient equine amplification, neurovirulence, and immune modulation. Specific aim 1: Identify genetic determinants of VEEV amplification and cell tropism using parental and chimeric strains of epizootic/enzootic subtype IE VEEV. As early cell targets of VEEV infection, equine macrophages/dendritic cells (DCs) will be tested for their ability to support viral replication. Equine peripheral blood leukocytes (PBLs) will also be examined for viral replication in vivo during early infection. Specific aim 2: Identify genetic determinants of VEEV pathogenicity and immunogenicity using parental and chimeric strains of epizootic/enzootic subtype IE VEEV. Infected equine macrophage/DC cultures will be tested for altered innate/adaptive immune responses using interferon (IFN) bioassays, quantitative RT-PCR of cytokines (including type I IFNs), and cDNA microarrays for global gene regulation. Neurologic symptoms will be quantitatively assessed in experimentally infected equids; neural/extraneural tissues will be qualitatively/quantitatively examined for gross/histopathologic lesions. Sera from infected animals will be measured for IFN induction, and PBLs and lymphoid/non-lymphoid tissues will be analyzed for altered gene expression of cytokines using quantitative RT-PCR. Collectively, these experiments will advance understanding of VEE emergence mechanisms and reveal natural determinants of amplification/virulence. Environment: Dr. Adams' project complements current, NIH-funded research of her proposed sponsor, Dr. Scott Weaver. The primary site of training, the University of Texas Medical Branch (UTMB), has superb facilities and equipment and outstanding faculty who study various aspects of arboviruses, including comparative medicine, immunology, and vaccine development. PUBLIC HEALTH RELEVANCE: Venezuelan equine encephalitis virus (VEEV) is the cause an important zoonotic disease that occurs throughout the Americas, with the demonstrated capacity to cause explosive outbreaks affecting hundreds-of- thousands of people and equids. Major epidemics in humans only occur when VEEV replicates to high titers in the blood of infected horses, donkeys, and mules, which is then transmitted to humans by infected mosquitoes. Using a VEEV isolate from a recent outbreak in Mexico, the purpose of the proposed study is identify viral genes that are responsible for efficient replication and neurovirulence in horses, which is critically important for the successful development of future intervention strategies (i.e., vaccines, therapeutics, and other human disease control measures).

描述（由申请人提供）：候选人：亚当斯博士在德克萨斯A&M大学以优异成绩完成了她的DVM，她完成了她的博士学位。在康奈尔大学的道格拉斯·安特扎克博士的实验室，研究胎儿逃避母体免疫反应的机制。亚当斯博士将运用她作为免疫学家的技能来追求她的长期目标，即了解甲病毒性脑炎的发病机制，这些知识最终将用于开发有效的疫苗/疗法。拟议的项目旨在弥合她从讲师到比较医学研究永久学术职位的过渡。 委内瑞拉马脑炎病毒（VEEV）是一种自然出现的虫媒病毒，威胁着整个美洲的人类健康。人类流行病仅发生在马类动物中VEEV有效扩增时。VEEV也被认为是一种潜在的生物恐怖分子制剂，被NIAID归类为B类优先病原体。最近爆发的流行亚型IE VEEV强调了VEE出现在人群中的持续威胁，以及我们对它在马科动物中扩增并通过突变改变毒力的能力所知甚少。本建议的目的是剖析的作用，结构和非结构蛋白的流行亚型IE VEEV引起有效的马扩增，神经毒力和免疫调节。具体目标1：使用流行/地方流行亚型IE VEEV的亲本和嵌合株鉴定VEEV扩增和细胞嗜性的遗传决定因素。作为VEEV感染的早期细胞靶标，将测试马巨噬细胞/树突状细胞（DC）支持病毒复制的能力。还将在早期感染期间检查马外周血白细胞（PBL）的体内病毒复制。具体目标2：使用流行/地方流行亚型IE VEEV的亲本和嵌合株鉴定VEEV致病性和免疫原性的遗传决定因素。将使用干扰素（IFN）生物测定、细胞因子（包括I型IFN）的定量RT-PCR和用于全局基因调控的cDNA微阵列检测感染的马巨噬细胞/DC培养物的先天性/适应性免疫应答改变。将在实验感染的马中定量评估神经症状;将定性/定量检查神经/神经组织的大体/组织病理学病变。将测量来自感染动物的血清的IFN诱导，并将使用定量RT-PCR分析PBL和淋巴/非淋巴组织的细胞因子的基因表达改变。总的来说，这些实验将促进对VEE出现机制的理解，并揭示扩增/毒力的自然决定因素。 环境：亚当斯博士的项目补充了她提议的赞助人斯科特·韦弗博士目前由NIH资助的研究。培训的主要地点是德克萨斯大学医学分支（UTMB），拥有一流的设施和设备以及研究虫媒病毒各个方面的优秀教师，包括比较医学，免疫学和疫苗开发。 公共卫生相关性：委内瑞拉马脑炎病毒（VEEV）是一种在整个美洲发生的重要人畜共患疾病，已证明能够引起影响数十万人和马的爆炸性爆发。只有当VEEV在受感染的马、驴和骡子的血液中复制到高滴度时，人类才会发生重大流行病，然后通过受感染的蚊子传播给人类。使用最近在墨西哥爆发的VEEV分离株，拟议研究的目的是确定负责马中有效复制和神经毒力的病毒基因，这对于成功开发未来干预策略至关重要（即，疫苗、治疗剂和其他人类疾病控制措施）。