Project 1: Definition of the structural principles underlying broadly protective humoral immunity to coronaviruses

项目 1：冠状病毒广泛保护性体液免疫的结构原理的定义

基本信息

批准号：
10425030
负责人：
David Veesler
金额：
$ 88.19万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-02 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10425030
关键词：
Animals Antibodies Antibody Response Antigens Binding Biological Assay COVID-19 pandemic COVID-19 vaccine Complex Coronavirus Coronavirus spike protein Cryoelectron Microscopy Development Distant Enzyme-Linked Immunosorbent Assay Epitopes Escape Mutant Glycoproteins Heterophile Antigens Human Humoral Immunities Immunity Immunize Macaca Macaca fascicularis Maps Mediating Memory B-Lymphocyte Merbecovirus Modeling Molecular Monoclonal Antibodies Mosaicism Mus Plasma Cells Resolution Sarbecovirus Serology test Specificity Structure Surface Vaccinated Vaccination Vaccine Design Vaccines Virus Work base betacoronavirus coronavirus vaccine cross reactivity design human coronavirus member mutation screening nanoparticle neutralizing antibody neutralizing monoclonal antibodies nonhuman primate pandemic preparedness polyclonal antibody protective efficacy rational design receptor binding response

项目摘要

PROJECT SUMMARY – PROJECT 1: Definition of the structural principles underlying broadly protective humoral immunity Although the COVID-19 pandemic has accelerated the development of SARS-CoV-2 vaccines at an unprecedented pace, no licensed vaccines elicit broad protection against a large spectrum of human coronaviruses. There is therefore an urgent need for vaccines inducing broad protection against currently circulating and distantly related betacoronaviruses for pandemic preparedness. The proposed Project aims to identify epitopes targeted by cross-reactive and broadly neutralizing anti-betacoronavirus antibodies to obtain an antigenic map of targets present at the surface of betacoronavirus spike trimers to guide our vaccine design efforts. Broadly neutralizing sarbecovirus antibodies recognizing the spike receptor-binding domain have recently been discovered, however, they do not cross-react with members of other subgenera. Previous studies have shown that the spike fusion machinery (S2 subunit), which is more conserved than the S1 subunit, harbors conserved epitopes targeted by cross-reactive polyclonal antibodies. Although a few β-coronavirus cross- reactive monoclonal antibodies are known, a deep understanding of the diversity of epitopes targeted by broadly neutralizing antibodies and their quantitative contribution to neutralization is lacking, thereby hindering the rational design of vaccines eliciting broad immunity. We will use three approaches to determine the molecular determinants of broad antibody-mediated coronavirus immunity by unveiling the types, specificities, and diversity of broadly neutralizing antibodies targeting all three main betacoronavirus subgenera (sarbecovirus, merbecovirus, and embecovirus). First, we will characterize the binding and neutralizing breath of polyclonal sera from nonhuman primates immunized with nanoparticle vaccines co-displaying multiple different RBD- and spike-based antigens. Second, we will determine the epitope specificities of cross-reactive antibodies in these sera using serological assays and by directly visualizing polyclonal antibodies in complex with vaccine-matched and heterologous antigens using cryo-electron microscopy. Finally, we will isolate monoclonal antibodies from nonhuman primates immunized with multivalent nanoparticle vaccines and characterize their structures at high resolution as well as their binding, neutralizing, and protective breadth.

项目摘要 - 项目1：广泛基础的结构原理的定义保护性体液免疫尽管COVID-19大流行已加速了SARS-COV-2疫苗在前所未有的空间，没有执照的疫苗可为大量人类提供广泛的保护冠状病毒。因此，迫切需要疫苗引起广泛的保护目前循环且相关的大流行准备。拟议的项目旨在通过交叉反应性和广泛中和抗抗核负纳病毒抗体靶向的表位以获得在Betacoronavirus尖峰夹具表面存在的靶标的抗原图，以引导我们的疫苗设计努力。识别尖峰受体结合结构域的广泛中和SARBECOVIRUS抗体具有但是，最近发现它们与其他子属的成员没有反应。先前的研究已经表明，尖峰融合机械（S2亚基）比S1亚基更保守由交叉反应多克隆抗体靶向的保守表位。虽然几个β-核纳病毒交叉已知反应性单克隆抗体，对广泛针对的表位的多样性有深刻的理解缺乏中和抗体及其对谈判的定量贡献，从而阻碍了疫苗的合理设计引起了广泛的免疫力。我们将使用三种方法来确定分子通过揭示类型，特异性和多样性的范围，广泛抗体介导的冠状病毒免疫的决定因素针对所有三个主要betacoronavirus subgenera的广泛中和抗体（SARBECOVIRUS， Merbecovirus和Embecovirus）。首先，我们将表征多克隆的结合和中和呼吸用纳米颗粒疫苗免疫的非人类隐私的血清共同播放多个不同的RBD和基于尖峰的抗原。其次，我们将确定其中的交叉反应性抗体的表位规范血清使用血清学分析，并直接可视化复合物与疫苗匹配的多克隆抗体使用冷冻电子显微镜和异源抗原。最后，我们将从用多价纳米颗粒疫苗免疫的非人类隐私，并在高处表征其结构分辨率及其结合，中和和受保护的广度。