Neutralization of Eastern equine encephalitis virus by human monoclonal antibodies

人单克隆抗体中和东方马脑炎病毒

• 批准号: 9757522
• 负责人: Lauren Williamson
• 金额: $ 2.97万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9757522
• 关键词: Address; Alanine; Alphavirus; American; Antibodies; Antibody Repertoire; Antibody Response; Antibody Specificity; Antigens; Antiviral Agents; Binding; Biological Assay; Bioterrorism; Capsid Proteins; Category B pathogen; Cells; Centers for Disease Control and Prevention (U.S.); Complex; Culicidae; Detection; Deterioration; Disease Outbreaks; Eastern Equine Encephalitis Virus; Electron Microscopy; Epitope Mapping; Epitopes; Equus caballus; Fellowship; Glycoproteins; Goals; Heterodimerization; Human; Human papillomavirus 16 E1 protein; Hybridomas; Immune; Immune response; Immune system; Immunity; Immunology; In Vitro; Infection; Knowledge; Laboratories; Libraries; Membrane; Molecular Conformation; Molecular Structure; Monoclonal Antibodies; Mus; Mutagenesis; Neurologic Signs; Neutralization Tests; Plaque Assay; Play; Prevention; Principal Investigator; Quantitative Reverse Transcriptase PCR; Recombinants; Recovery; Reporting; Research; Resolution; Role; Scanning; Scientist; Structural Protein; Structure; Surface; Survivors; Techniques; Technology; Testing; Therapeutic; Training; Transmembrane Domain; United States National Institutes of Health; Vaccines; Viral; Virion; Virulent; Virus; Virus Diseases; Virus Replication; X-Ray Crystallography; career; clinically relevant; clinically significant; crosslink; design; experience; feeding; human monoclonal antibodies; human mortality; insight; mortality; murine antibody; neutralizing antibody; neutralizing monoclonal antibodies; particle; prevent; programs; protein structure; prototype; receptor; stoichiometry; structural biology; virology

PROJECT SUMMARY/ABSTRACT The overall goal of this project addresses the molecular and structural basis of neutralization utilized by human monoclonal antibodies (mAbs) against Eastern equine encephalitis virus (EEEV). The North American lineage of EEEV (NA-EEEV) is the most virulent of the encephalitic alphaviruses with up to 70% human and 90% equine mortality rate. In addition, of those that survive infection, up to 80% show signs of neurological deterioration. On average, eight human cases a year in the US are reported. However, the recent rise in detection of NA-EEEV in human-feeding mosquito species raises concern for a large outbreak in the eastern US. Moreover, NA-EEEV is considered a NIH Category B priority pathogen and USDA/CDC Select Agent due to its potential threat as a bioterrorism agent. There are no approved antiviral drugs or licensed human vaccines available for NA-EEEV. The antibody response to alphaviruses has been shown to be an important part of the immune response in conferring protective immunity and aiding in the clearance and recovery from infection. However, the fundamental molecular and structural mechanisms of action of antibodies in humans to NA-EEEV remain poorly defined. To fill this gap in knowledge, the objective of this study is to determine the mechanisms by which human mAbs neutralize NA-EEEV. The overarching hypothesis of this study is that the principle mode of action of human neutralizing mAbs is to stabilize the virus particle and inhibit viral fusion to host cells. To test this hypothesis, I will isolate human mAbs from naturally infected survivors of NA-EEEV to the BSL-2 chimeric virus, Sindbis/EEEV (SINV/NA-EEEV) and recombinant NA-EEEV structural proteins. I will characterize the panel of human mAbs isolated for specific reactivity and neutralization potency against SINV/NA-EEEV. Of the human mAbs that neutralize NA-EEEV, I then will determine the step(s) in the replication cycle the mAbs neutralize the virus through in vitro mechanistic assays. To identify neutralizing antigenic determinants, I will use epitope mapping, alanine scanning mutagenesis, and structural biology techniques. Determination of the neutralizing antigenic determinants recognized by human NA-EEEV mAbs will inform how mAbs interact with NA-EEEV. The results obtained from this project will fill major gaps in knowledge about the human antibody response to NA-EEEV, will identify potential correlates of protection, and may facilitate the design of efficient therapeutics and vaccines against this clinically relevant alphavirus. The proposal outlines a comprehensive fellowship training plan that will prepare me for a high-impact career in the field of viral immunology. To accomplish this program and research, we have assembled a collaborative team consisting of principal investigators and staff scientists with exceptional expertise in the immunology, virology, and structural biology fields to provide support and guidance for this study.

项目摘要/摘要 该项目的总体目标是针对人类使用的中和的分子和结构基础 针对东马脑炎病毒（EEEV）的单克隆抗体（mAb）。北美血统 EEEV（NA-EEEV）是最有毒的脑α病毒，最多70％，90％ 马死亡率。此外，在幸存感染的患者中，多达80％的神经系统迹象 恶化。平均而言，美国每年有八例人类案件。但是，最近的增长 在人类喂养蚊子中发现Na-Eeev的发现引起了人们对东部大规模爆发的关注 我们。此外，NA-EEEV被认为是NIH B类优先级病原体和USDA/CDC选择代理 作为生物恐怖主义者的潜在威胁。没有批准的抗病毒药或持牌人 可用于NA-EEEV的疫苗。对α病毒的抗体反应已被证明是重要的 免疫反应的一部分在赋予保护性免疫和帮助清除和从中恢复的一部分 感染。但是，人类抗体的基本分子和结构机制 NA-EEEV的定义较差。为了填补这一知识的差距，这项研究的目的是确定 人mab中和na-eeev的机制。这项研究的总体假设是 人类中和mAb的主要作用方式是稳定病毒颗粒并抑制病毒融合到 宿主细胞。为了检验这一假设，我将将人物mAb从Na-Eeev的自然感染幸存者中隔离到 BSL-2嵌合病毒，Sindbis/EEEV（SINV/NA-EEEV）和重组Na-EEEV结构蛋白。我会 表征分离出的人物板，以特异性反应性和中和效力 SINV/NA-EEEV。在中和na-eeev的人mab中，我将确定 复制周期mAb通过体外机械测定法中和病毒。识别中和 抗原决定因素，我将使用表位图，丙氨酸扫描诱变和结构生物学 技术。确定人Na-Eeev mAb识别的中和抗原决定因素 将告知mab如何与Na-Eeev互动。从该项目获得的结果将填补主要空白 关于人类抗体对NA-EEEV的反应的知识将确定保护的潜在相关性，并且 可以促进针对这种临床相关α病毒的有效治疗和疫苗的设计。这 提案概述了一项全面的奖学金培训计划，这将使我为我的高影响事业做好准备 病毒免疫学领域。为了完成该计划和研究，我们组建了一个协作团队 由主要研究人员和员工科学家组成，在免疫学，病毒学， 和结构生物学领域，为这项研究提供支持和指导。