Structural Determinants of Human Antibodies neutralizing the Ebola Virus

中和埃博拉病毒的人类抗体的结构决定因素

• 批准号: 9304960
• 负责人: Jens Meiler
• 金额: $ 23.54万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9304960
• 关键词: Affinity; Algorithms; Antibodies; Antibody Repertoire; Antigen-Antibody Complex; Binding; Biological; Central Africa; Collaborations; Communities; Complement; Complex; Computational Biology; Computer Simulation; Crystallization; Data; Databases; Development; Disease Outbreaks; Ebola virus; Electron Microscopy; Engineering; Europe; Family; Filoviridae; Filovirus; Frankfurt-Marburg Syndrome Virus; Geography; Glycoproteins; Goals; Human; Hybrids; Immune; Immune response; Infection; Influenza; Laboratories; Letters; Maps; Membrane Glycoproteins; Methods; Modeling; Modernization; Mutation; Positioning Attribute; Procedures; Property; Proteins; Protocols documentation; Publications; Research; Research Personnel; Resolution; Somatic Mutation; Structure; System; Technology; Testing; Therapeutic antibodies; Time; Uganda; Universities; Vaccination; Vaccines; Viral; Viral Hemorrhagic Fevers; Virus; Western Africa; Work; data management; density; design; follow-up; high risk; improved; innovation; member; neutralizing antibody; next generation sequencing; novel; structural biology; therapeutic development; therapeutic vaccine; vaccination strategy; ward

SUMMARY The filovirus family includes Marburg and Ebola viruses, most of which cause highly lethal hemorrhagic fever. The first filovirus was identified when it sickened laboratory workers in Europe in 1967. Since then, filo- viruses have re-emerged multiple times, with modern strains conferring greater lethality (~90%). Ebola virus is typically found in Central Africa, but re-emerged in Western Africa in 2014 to cause an on-going outbreak un- precedented in magnitude and geographic spread that has already claimed the lives of thousands of people. There is also a current outbreak of Marburg virus in Uganda. Last month structural data on several neutralizing antibodies (Abs) in complex with the viral target enve- lope surface glycoprotein (GP) became available to us, enabling for the first time the research proposed within this PAPID proposal. Specifically, co-crystal structures of one Ab with the Marburg and Ebola GP, respectively, as well as electron microscopy (EM) density maps for six additional Abs. It is our Goal to investigate the shared structural determinants of human Abs neutralizing the filovirus. While the present study thereby focuses on the biological question of the immune response to an infection by the Ebola and Marburg viruses, it has the potential to facilitate development of therapeutic strategies by others. Within the scope of this R21 proposal we pursue three goals: Aim 1 creates atomic detail models of the Ab/GP interface for all seven Abs with available EM density maps. We will use the co-crystal structure to test our computational protocol, and then apply it to the remaining six cases. As a result, we will obtain a map of critical structural determinants an Ab needs to fulfill for being neutralizing. Aim 2 will identify novel Abs from the repertoire of naïve subjects that are likely to neutralize Ebola and/or Marburg viruses with a limited set of muta- tions. We also will compare the Ab repertoires of filovirus-naïve (i.e. not previously infected) humans to those of immune (previously infected) subjects available to us. These Abs from naïve subjects are important, as they could evolve into neutralizing Abs upon infection or vaccination. We will redesign these Abs computationally to bind and neutralize Ebola and/or Marburg viruses, to increase the pool of known neutralizing Abs, paving the way for a successful vaccination strategy. In Aim 3 we will characterize these Abs experimentally for binding and neutralization activity. While ultimately a structural characterization of the most promising of these Abs is planned, it is outside the scope of the R21 proposal.

概括 丝状病毒家族包括马堡和埃博拉病毒，其中大多数引起高致命的出血性 发烧。 1967年，它在欧洲生病时发现了第一个丝状病毒。从那时起， 病毒已经重新出现了多次，现代菌株会议更大的致死性（约90％）。埃博拉病毒是 通常在中非发现，但在2014年在西非重新出现，导致持续的爆发 - 在规模和地理传播中，已经夺走了成千上万人的生命。 目前，乌干达也有马尔堡病毒爆发。 上个月，关于与病毒靶的复合物中的几种中和抗体（ABS）的结构数据 我们可为我们提供LOPE表面糖蛋白（GP），这是首次提出的研究 这个纸质提案。具体而言，一个AB的共晶结构分别与Marburg和Ebola GP 以及电子显微镜（EM）密度图，用于六个额外的ABS。我们的目标是调查共享 人类ABS的结构决定剂中和丝状病毒。而本研究则重点 关于埃博拉病毒感染的免疫反应的生物学问题，它具有 促进他人制定治疗策略的潜力。 在此R21提案的范围内，我们实现了三个目标：AIM 1创建原子细节模型 所有七个ABS的AB/GP接口，带有可用的EM密度图。我们将使用共晶结构进行测试 我们的计算协议，然后将其应用于其余六种情况。结果，我们将获得一张地图 临界结构决定者AB需要实现中和。 AIM 2将从 幼稚的受试者的曲目​​可能会中和埃博拉病毒和/或马堡病毒，其中一组有限的muta- tions。我们还将比较Filevirus-Noïve（即以前未感染）人类的AB曲目与 免疫（以前感染）受试者可供我们使用。这些来自天真主题的腹肌很重要，因为 在感染或疫苗接种后可以演变成中和ABS。我们将通过计算将这些ABS重新设计为 结合并中和埃博拉病毒和/或马堡病毒，增加已知中和的ABS的库，铺路 成功进行疫苗接种策略的方式。在AIM 3中，我们将在实验上表征这些ABS的结合和 中和活性。虽然最终对这些ABS最有希望的结构表征是 计划，它不在R21提案的范围之内。