flowBEAT: a multiparameter flow cytometry assay to discover longitudinal antibody responses associated with efficient recovery from COVID-19

flowBEAT：一种多参数流式细胞术检测，用于发现与 COVID-19 有效恢复相关的纵向抗体反应

• 批准号: 10355136
• 负责人: Eliver Ghosn
• 金额: $ 23.22万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10355136
• 关键词: 2019-nCoV; Address; Animal Model; Antibodies; Antibody Response; Antibody Specificity; Antibody titer measurement; Biological Assay; COVID-19; COVID-19 mortality; COVID-19 patient; COVID-19 severity; Cells; Coronavirus; Data; Disease; Disease Outcome; Flow Cytometry; Fluorescence; Future; HIV; Human; IgA1; IgA2; IgE; IgG1; IgG2; IgG3; IgG4; Immune response; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Interferon Type I; Interferons; Knowledge; Label; Longevity; Longitudinal Studies; Measurement; Measures; Methods; Microspheres; Nonstructural Protein; Nucleocapsid Proteins; Pathogenesis; Patients; Prevalence; Proteins; Recovery; Research Personnel; Role; SARS-CoV-2 spike protein; SARS-CoV-2 variant; Sampling; Serology; Serology test; Serum; Severity of illness; Specificity; Symptoms; System; Technology; Variant; Viral Proteins; Virus; antigen detection; base; cohort; falls; fighting; flexibility; high dimensionality; human coronavirus; immunoregulation; improved; light scattering; multiplex assay; mutant; new technology; novel; pandemic disease; prevent; receptor binding; response; severe COVID-19; vaccine development; virtual

Project Summary Mounting an effective antibody response against SARS-CoV-2 is key to fight off the infection. However, not all antibodies produced against the virus are protective. Recent studies show that high titers of immunoglobulin G (IgG) against the SARS-CoV-2 Spike (S) protein during natural infection correlate with disease severity. The S protein, highlighted due to its receptor-binding role in host cell infection, represents only one of potentially 29 proteins that are encoded by the SARS-CoV-2 virus. In fact, recent studies show that SARS-CoV- 2 non-structural proteins, including ORF3b, ORF8, ORF9b, and ORF9c, have the ability to suppress host antiviral type-I interferon (IFN) and elicit sustained antibody responses. These findings raise the question of whether antibodies produced against these immunomodulatory viral proteins early during the infection could provide protection against severe COVID-19. In addition to the antibody specificity to different SARS-CoV-2 proteins, the antibody isotype and subclass can also influence the disease outcome. However, much of the serology data from SARS-CoV-2- infected patients mainly focus on IgG and IgM responses to S and nucleocapsid proteins only. In this proposal, we aim to tackle this problem by developing a new technology that will allow for simultaneous measurement of 8 antibody isotypes and subclasses (IgM, IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgE) against 24 SARS-CoV-2 and endemic HCoV-encoded proteins, including against proteins from emerging virus variants, all in a single multiplexed assay (Aim 1). We will apply this technology, namely flowBEAT (flow cytometry-based BEads assay to detect Antigen-specific antibody isoTypes), to 600 serum samples collected from a longitudinal study (6 timepoints; ≥ 6 months) of a well-characterized cohort of 100 COVID-19 patients presenting with either mild or severe symptoms (Aim 2). We hypothesize that mild, but not severe, COVID-19 patients will show an early and sustained antibody subclass against immunomodulatory viral proteins, particularly against viral proteins known to suppress IFN responses. We expect this study to provide an in-depth view of the breadth (type, specificity, and longevity) of antibody responses mounted against 24 SARS-CoV-2- and HCoV-encoded proteins that are associated with efficient recovery from COVID-19. Our study can also inform on ongoing and future vaccine development by identifying proteins that generate long-lasting and protective antibody subclasses in the recovered patients. Finally, the flexible modular feature of flowBEAT can quickly be adapted to study the breadth of antibody responses in virtually any disease.

项目概要 针对 SARS-CoV-2 建立有效的抗体反应是抵抗感染的关键。 最近的研究表明，并非所有针对该病毒的抗体都具有保护作用。 自然感染期间针对 SARS-CoV-2 Spike (S) 蛋白的免疫球蛋白 G (IgG) 与疾病相关 S 蛋白因其在宿主细胞感染中的受体结合作用而突出，仅代表其中之一。 SARS-CoV-2 病毒编码的潜在 29 种蛋白质事实上，最近的研究表明 SARS-CoV- 2种非结构蛋白，包括ORF3b、ORF8、ORF9b和ORF9c，具有抑制宿主抗病毒的能力 I 型干扰素 (IFN) 并引发持续的抗体反应，这些发现提出了一个问题： 在感染早期针对这些免疫调节病毒蛋白产生的抗体可以提供 预防严重的 COVID-19。 除了针对不同 SARS-CoV-2 蛋白的抗体特异性外，抗体同种型和 亚类也会影响疾病结果，然而，大部分血清学数据来自 SARS-CoV-2-。 感染患者主要关注对 S 和核衣壳蛋白的 IgG 和 IgM 反应。 我们的目标是通过开发一种新技术来解决这个问题，该技术将允许同时测量 针对 24 种 SARS-CoV-2 的 8 种抗体同种型和亚类（IgM、IgG1、IgG2、IgG3、IgG4、IgA1、IgA2、IgE） 和地方性 HCoV 编码的蛋白质，包括针对新出现的病毒变种的蛋白质，全部都在一个单一的 我们将应用这项技术，即 flowBEAT（基于流式细胞术的 BEads）。 检测抗原特异性抗体 isoTypes），从纵向研究中收集 600 个血清样本 （6 个时间点；≥ 6 个月）由 100 名表现出轻度症状的 COVID-19 患者组成的队列 或严重症状（目标 2），轻度但不严重的 COVID-19 患者会出现早期症状。 和针对免疫调节病毒蛋白，特别是针对病毒蛋白的持续抗体亚类 已知可抑制 IFN 反应。 我们希望这项研究能够深入了解疾病的广度（类型、特异性和寿命） 针对 24 种 SARS-CoV-2 和 HCoV 编码蛋白的抗体反应，这些蛋白与 我们的研究还可以为当前和未来的疫苗开发提供信息。 鉴定在康复患者体内产生持久和保护性抗体亚类的蛋白质。 最后，flowBEAT 灵活的模块化功能可以快速适应抗体的广度研究 几乎对任何疾病都有反应。