Core C: Immunoglobulin Proteomics in COVID-19

摘要 研究核心C将成为COVID-19研究中血清学抗体库检查的组成部分 受试者以及从幸存者获得的恢复期血浆中。核心C将采用一套独特的 实验技术，包括用于鉴定单克隆抗体的蛋白质组学方法（Ig-seq） 序列，并用于确定分泌组分免疫球蛋白的表位特异性和功能 (IgG和伊加），其包含对SARS-CoV-2的多克隆应答。UT核心将检查 作为时间函数的个体血清mAb克隆型以及伊加和IgG之间的关系 在循环和BAL的剧目。到目前为止，已经分离出了所有人SARS-CoV-2反应性抗体， 仅来自外周中短暂循环的B细胞。丰富性、耐久性和 血清学抗体的相互联系-IgG与伊加，中和与非中和，粘膜与 系统-将变得清晰，因为这个研究核心全面分析的分子组成， 抗病毒IgG和伊加，并追踪它们产生的B细胞亚群。将启用这些研究 通过强大的样本收集从北查佩尔山和德州奥斯汀。我们预计， 研究核心C的结果将阐明（i）组成IgG的抗体宽度和效力的程度 和伊加在血清学谱系中的作用以及在保护中的作用（通过病毒阻断或通过Fc依赖性 机制）在小鼠模型中对抗地方病和人畜共患冠状病毒（项目1），以及（ii）详细的 在COVID-19临床中用于输血治疗的恢复期血浆的分子水平表征 试验（项目2）。应进行比较分析，以确定适应性免疫特征， 解释无症状、少症状和严重疾病之间的差异模式。这项研究将是 这对我们理解人类抗SARS-CoV-2抗体反应的性质和复杂性至关重要 大流行的祸害。

Abstract Research Core C will be integral to the examination of serological antibody repertoires in COVID-19 study subjects as well as in convalescent plasmas obtained from survivors. Core C will employ a set of unique experimental techniques including a proteomic methodology (Ig-seq) for the identification of the monoclonal sequences and for determining the epitope specificity and function of the secreted component immunoglobulins (IgG and IgA) comprising the polyclonal response to SARS-CoV-2. The UT Core will examine the persistence of individual serum mAb clonotypes as a function of time and also the relationships between the IgA and IgG repertoires in circulation and in BAL. To date, all human SARS-CoV-2-reactive antibodies have been isolated exclusively from B cells transiently circulating in the periphery. The significance of the abundance, durability, and interconnectivity of serological antibodies—IgG versus IgA, neutralizing versus non-neutralizing, mucosal versus systemic—shall become clear as this research core comprehensively analyzes the molecular composition of anti-viral IgG and IgA and traces the B-cell subpopulations from which they arose. These studies will be enabled by robust sample collections from UNC Chapel Hill and from UT Austin. We expect that the experimental outcomes of Research Core C will clarify (i) the extent of antibody breadth and potency of the constituent IgG and IgA in the serological repertoires and role in protection (via viral blockade or through Fc dependent mechanisms) against endemic and zoonotic coronaviruses in mouse models (Project 1), and (ii) the detailed molecular-level characterization of convalescent plasmas used for transfusion therapy in a COVID-19 clinical trial (Project 2). Comparative analyses shall be performed to determine adaptive immune signatures which may explain differential patterns among asymptomatic, oligosymptomatic, and severe disease. This research will be critical to our understanding the nature and complexity of human antibody responses against the SARS-CoV-2 pandemic scourge.