Antibody repertoire characterization in the context of coronaviruses

项目摘要。SARS-CoV-2，或2019年新型冠状病毒，是一种重大的大流行威胁，已经 截至2020年3月，导致数十万确诊病例和数万人死亡。 制定预防和治疗措施，以抵消正在进行的和任何未来的 因此，冠状病毒大流行对全世界的公共卫生具有极其重要的意义。S蛋白是 冠状病毒(CoV)的免疫优势区，由免疫系统识别并作为靶标 一系列中和抗体。被动转移中和抗体已被证明可以预防 冠状病毒感染动物模型。此外，工程化的灌流稳定的S蛋白免疫原已经 在MERS的背景下，已被证明在动物模型中诱导高滴度的冠状病毒中和抗体。 综上所述，这项先前的工作为确定和表征 SARS-CoV-2背景下的中和抗体。更广泛地说，更好地理解人类 对SARS-CoV-2 S蛋白以及其他相关CoV成员的抗体反应有助于了解 治疗性抗体优化和加速疫苗设计工作。 我们实验室最近开发了Libra-seq技术(将B细胞受体与抗原联系起来 通过测序的特异性)用于抗体发现和抗原特异性抗体的表征 剧目。与其他B细胞方法不同，Libra-seq是第一个能够同时检测 大量B细胞对理论上无限数量B细胞的bcr序列和抗原特异性 不同的抗原，在单细胞水平上。因此，Libra-seq提供了一个独特的机会来描述 识别SARS-CoV-2和其他病毒S蛋白的抗体类型和特异性 冠状病毒。 在这里，我们建议在SARS-CoV-2的背景下利用Libra-seq技术，主要有两个方面 目的：(1)鉴定识别人类多种抗原变异体的交叉反应抗体 冠状病毒感染，包括SARS-CoV-2、SARS-CoV-1和MERS-CoV，以及(2)评估 目前领先的冠状病毒候选疫苗与来自健康个体的抗体库相结合。 综上所述，这一应用中提出的努力将具有很高的翻译/临床潜力 对SARS-CoV-2和其他具有生物医学意义的冠状病毒病原体的影响。抗体谱系的类型 我们在这里建议开发的特征也很容易推广到其他病原体，并且作为 这将对制定既有和持久的反措施产生广泛和持久的影响。 新出现的传染病。

Project Summary. SARS-CoV-2, or the 2019 novel coronavirus, is a significant pandemic threat that has resulted in hundreds of thousands of diagnosed cases and tens of thousands of mortalities as of March 2020. The development of preventive and therapeutic measures that can counteract the ongoing, and any future, coronavirus pandemics is therefore of utmost significance for public health worldwide. The S protein is the immunodominant region of coronaviruses (CoV) recognized by the immune system and serves as the target for a number of neutralizing antibodies. Passive transfer of neutralizing antibodies has been shown to prevent coronavirus infection in animal models. Further, engineered prefusion-stabilized S protein immunogens have been shown to elicit high titers of coronavirus-neutralizing antibodies in animal models, in the context of MERS. Together, this prior work establishes a strong premise for targeting the identification and characterization of neutralizing antibodies in the context of SARS-CoV-2. More generally, a better understanding of the human antibody response to the S protein of SARS-CoV-2 as well as other related CoV members can help inform therapeutic antibody optimization and accelerate vaccine design efforts. Our laboratory recently developed the LIBRA-seq technology (LInking B-cell Receptor to Antigen specificity through sequencing) for antibody discovery and characterization of antigen-specific antibody repertoires. Unlike other B cell approaches, LIBRA-seq is the first to enable the simultaneous determination of BCR sequence and antigen specificity for a large number of B cells against a theoretically unlimited number of diverse antigens, at the single-cell level. LIBRA-seq therefore provides a unique opportunity for characterizing the types and specificities of antibodies that can recognize the S protein from SARS-CoV-2, as well as other CoV viruses. Here, we propose to utilize the LIBRA-seq technology in the context of SARS-CoV-2, with two major goals: (1) To identify cross-reactive antibodies that recognize multiple antigen variants associated with human coronavirus infection, including SARS-CoV-2, SARS-CoV-1, and MERS-CoV, and (2) To evaluate the ability of current lead CoV vaccine candidates to engage with antibody repertoires from healthy individuals. Taken together, the efforts proposed in this application will be of high potential translational/clinical impact for SARS-CoV-2 and other CoV pathogens of biomedical significance. The types of antibody repertoire characterization that we propose to develop here will also be readily generalizable to other pathogens, and as such, will have a broad and lasting impact on the development of countermeasures for established and emerging infectious diseases.