Structural Characterization of Coronavirus Antibodies Raised by Infection and Vaccination

项目3：摘要/摘要 SARS-CoV-2是一种新出现的肉瘤病毒属贝塔冠状病毒，于 2020年，感染数百万人，引发新冠肺炎病。另外两种人畜共患的贝塔冠状病毒，SARS-CoV(a MERS冠状病毒(一种美沙贝病毒)和MERS冠状病毒(一种汞病毒)在过去20年中也有暴发。类似SARS的疾病 病毒在蝙蝠体内传播，并对居住在蝙蝠携带不同病毒的洞穴附近的人进行血清学监测 冠状病毒显示可直接传播具有大流行潜力的SARS类病毒，这表明 需要冠状病毒疫苗。在项目3中，比约克曼实验室将使用结构生物学和生化 了解SARS冠状病毒在人体内诱导的抗体中和机制的方法 2感染或接种疫苗以及在实验动物中通过免疫接种。在目标1中，使用单抗Fabs的3D结构 与冠状病毒尖峰(S)三聚体结合，我们将推导出中和/结合的结构相关性 Nussenzweig博士在项目1中从(I)人类中分离的单抗和多克隆血浆 在~1个月后感染SARS-CoV-2，(Ii)感染后半至两年分离的成熟人抗体，(Iii)人 用莫德纳疫苗接种针对SARS-CoV-2的疫苗，以及(Iv)用候选疫苗接种动物 在Bieniasz和HatziioAnnou博士的项目2中开发，或在比约克曼的本项目的目标2中开发 实验室。该项目的目标2将跟进我们实验室对交叉反应抗体的可能性的评估 对肉瘤病毒的反应，我们制作了呈现受体结合的同型纳米颗粒 仅SARS-CoV-2的结构域(RBD)或与动物的RBD共同显示SARS-CoV-2RBD 贝塔冠状病毒(镶嵌纳米颗粒；4-8个不同的RBD)。通过结合抗体的功能分析结果 与赖斯博士合作，通过假型和正品病毒中和试验得出的中和结果 通过对注射RBD纳米颗粒的小鼠分离出的抗体的结构分析，我们将提纯我们的纳米颗粒 候选疫苗(S)，以增加其抵御肉瘤病毒的潜力。此外，我们还将发展 其他针对MerbecoVirus和/或AlphacoronaVirus的候选疫苗将在 赖斯博士和/或我们的NIH合作者在动物保护模型中使用了最好的泛肉瘤病毒疫苗， 文森特·蒙斯特博士。这个高度整合的项目有可能为创造疫苗做出贡献(S) 可以避免未来的全球大流行。

Project 3: Summary/Abstract SARS-CoV-2, a newly-emergent betacoronavirus in the sarbecovirus genus, resulted in a global pandemic in 2020, infecting millions and causing COVID-19 disease. Two other zoonotic betacoronaviruses, SARS-CoV (a sarbecovirus) and MERS-CoV (a merbecovirus), also resulted in outbreaks within the last 20 years. SARS-like viruses circulate in bats and serological surveillance of people living near caves where bats carry diverse coronaviruses demonstrate direct transmission of SARS-like viruses with pandemic potential, suggesting a pan- coronavirus vaccine is needed. In Project 3, the Bjorkman lab will use structural biology and biochemical approaches to understand the neutralization mechanisms of antibodies (Abs) elicited in humans by SARS-CoV- 2 infection or vaccination and in experimental animals by immunization. In Aim 1, using 3D structures of Ab Fabs bound to coronavirus spike (S) trimers, we will derive the structural correlates of neutralization/binding for monoclonal Abs (mAbs) and polyclonal plasmas isolated in Project 1 by Dr. Nussenzweig from (i) humans infected by SARS-CoV-2 after ~1 month, (ii) matured human Abs isolated ½ - 2 years after infection, (iii) humans vaccinated against SARS-CoV-2 with the Moderna vaccine, and (iv) animals immunized with vaccine candidates developed in Project 2 by Drs. Bieniasz and Hatziioannou or developed in this project's Aim 2 in the Bjorkman laboratory. Aim 2 of this project will follow up on our lab's evaluations of the potential for cross-reactive antibody responses to sarbecoviruses, for which we made homotypic nanoparticles presenting the receptor-binding domain (RBD) of only SARS-CoV-2 or co-displaying SARS-CoV-2 RBD along with RBDs from animal betacoronaviruses (mosaic nanoparticles; 4-8 distinct RBDs). By combining results of functional analyses of Ab neutralization derived from pseudotyped and authentic virus neutralization assays in collaboration with Dr. Rice with structural analyses of Abs isolated from RBD-nanoparticle–injected mice, we will refine our nanoparticle vaccine candidate(s) to increase their potential to protect against sarbecoviruses. Furthermore, we will develop additional vaccine candidates against merbecoviruses and/or alphacoronaviruses that will be evaluated along with the best pan-sarbecovirus vaccine in animal models of protection by Dr. Rice and/or our NIH collaborator, Dr. Vincent Munster. This highly-integrated project has the potential to contribute to creation of vaccine(s) that could avert future global pandemics.