COVID19: Anti-SARS-CoV-2 Antibodies and Infection Severity

COVID19：抗 SARS-CoV-2 抗体和感染严重程度

• 批准号: 10152299
• 负责人: KENNETH M KAUFMAN
• 金额: --
• 依托单位: CINCINNATI VA MEDICAL CENTER RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10152299
• 关键词: 2019-nCoV; Amino Acids; Antibodies; Antibody Response; Antibody Specificity; Antigenic Specificity; Antigens; Asphyxia; B-Lymphocyte Epitopes; Behavior; Biological Assay; Breathing; COVID-19; COVID-19 mortality; COVID-19 pandemic; Cessation of life; Characteristics; Clinical; Contracts; Coronavirus; Dangerousness; Development; Diagnostic tests; Disease; Disease Outcome; Disease model; Donor Selection; Enzyme-Linked Immunosorbent Assay; Epidemiology; Epitope spreading; Epitopes; Formulation; Functional disorder; Healthcare Systems; Human; Human Herpesvirus 4; Immune response; Individual; Infection; Infectious Agent; Inflammatory Response; Knowledge; Ku70 protein; Learning; Libraries; Lung diseases; Lupus; Maps; Mediating; Middle East Respiratory Syndrome; Minor; Morbidity - disease rate; Occupations; Outcome; Passive Immunotherapy; Pathogenicity; Patient Care; Patients; Peptides; Phage Display; Phase; Plasma; Polyethylenes; Population; Property; Proteins; Public Health; Reagent; Recombinant Proteins; Research Personnel; Resolution; Risk; SARS coronavirus; SARS-CoV-2 antibody; SARS-CoV-2 antigen; SARS-CoV-2 infection; Scourge; Severe Acute Respiratory Syndrome; Severities; Severity of illness; Solid; Source; Specificity; Structure; Techniques; Testing; Therapeutic; Time; Vaccination; Vaccine Design; Veterans; Virus; Western Blotting; Work; base; biosafety level 3 facility; comorbidity; experience; experimental study; glycosylation; lung injury; military veteran; older men; pandemic disease; prevent; theories; therapy design; vaccine response

The new Covid-19 virus is deadly for 0.2% to 4% of people infected, with older men the most severely afflicted with this scourge, being mortal to ~15% of those infected over 80 years, making the US Veteran population especially vulnerable. It is critically important to understand the mechanisms of this lethal pathophysiology in order to design interventions that will save lives. MERS, SARS-CoV, and now the new human Covid-19 mean that three coronaviruses have threatened to become horrific public health problems for the entire world; this last and most recent one, Covid-19, has succeeded. Some of the antibodies formed against this virus appear to accentuate the clinical severity of disease, possibly making the specific details of the immune response responsible for the demise of many infected patients. We have had more than two decades of experience as VA Merit supported investigators exploring the antigenic structure of the lupus autoantigens, leading us to the theory that Epstein-Barr virus causes lupus, which is a hypothesis now supported by convincing circumstantial evidence. We propose to apply our experience to exploring the details of the fine antigenic specificity of the anti-Covid-19 antibody response in order to understand what specific antibodies are dangerous. We propose to explore two hypotheses: Hypothesis 1. Specific and detailed knowledge of antibody immune responses to Covid-19 will be useful for elucidating pathogenic mechanisms, predicting disease severity, designing vaccines, selecting therapeutic plasmas, making patient care decisions, informing population epidemiology, determining infection dynamics, and choosing individual behaviors in the face of the Covid-19 pandemic. Hypothesis 2. Anti-Covid-19 antibody fine antigenic specificity will correlate with clinical manifestations and infection severity. We propose two aims: Aim 1. Develop and apply assays to dissect the antigenic fine specificity of antibodies forming against shared and sub-strain specific Covid-19 antigens in the natural human Covid-19 infection. Aim 2. Isolate and purify anti-Covid-19 antibodies against the major epitopes and any specific antibodies that are associated with severe pulmonary disease. We will use Western blotting, solid phase assays (ELISAs), phage display libraries expressing 30-mer amino acid peptides, and synthesized overlapping fmoc peptides to fully characterize the fine specificity antibody response to the Covid-19 virus. We will obtain plasma and serum from Veterans and others which we will evaluate for correlates with clinical outcome. We will isolate individual specificities and characterize their properties with respect to type, isotype, glycosylation, and activity. We will perform pseudotype assays on whole repertoires and individual antibody specificities in BSL2 and test for confirmation with neutralization with collaborators with BSL3 facilities. From these studies we hope to learn how to identify Veterans who are likely to have a more severe illness and provide clues for developing approaches that could succeed in ameliorating infection severity and prevent severe disease.

新的 Covid-19 病毒对 0.2% 至 4% 的感染者来说是致命的，其中老年男性受影响最严重 由于这种祸害，80 年来约有 15% 的感染者死亡，这使得美国退伍军人人口 特别脆弱。了解这种致命的病理生理学机制至关重要 以便设计能够拯救生命的干预措施。 MERS、SARS-CoV 以及现在新的人类 Covid-19 意味着 三种冠状病毒有可能成为全世界可怕的公共卫生问题；这 最后一个也是最近的一个，Covid-19，已经成功了。 针对这种病毒形成的一些抗体似乎加剧了疾病的临床严重性，可能是 免疫反应的具体细节导致了许多感染患者的死亡。我们 拥有二十多年作为 VA Merit 支持研究人员探索抗原的经验 狼疮自身抗原的结构，使我们得出 Epstein-Barr 病毒引起狼疮的理论，这是一种 假设现在得到了令人信服的间接证据的支持。我们建议将我们的经验应用于 探索抗 Covid-19 抗体反应的精细抗原特异性的细节，以便 了解哪些特定抗体是危险的。我们建议探索两个假设： 假设 1. 对 Covid-19 抗体免疫反应的具体而详细的了解将是 对于阐明致病机制、预测疾病严重程度、设计疫苗、 选择治疗血浆，做出患者护理决定，告知人群流行病学， 确定感染动态并选择面对 Covid-19 的个人行为 大流行。 假设 2. 抗 Covid-19 抗体良好的抗原特异性将与临床相关 表现和感染严重程度。 我们提出两个目标： 目标 1. 开发并应用分析方法来剖析针对共享抗原形成的抗体的抗原精细特异性。 以及自然人类 Covid-19 感染中的亚株特异性 Covid-19 抗原。 目标 2. 分离和纯化针对主要表位的抗 Covid-19 抗体以及任何特定抗体 与严重的肺部疾病有关。 我们将使用蛋白质印迹、固相测定 (ELISA)、表达 30 聚体氨基的噬菌体展示文库 酸性肽，并合成重叠的 fmoc 肽，以充分表征精细特异性抗体 对 Covid-19 病毒的反应。我们将从退伍军人和其他人那里获得血浆和血清 评估与临床结果的相关性。我们将分离出个体的特殊性并描述他们的特征 关于类型、同种型、糖基化和活性的特性。我们将进行假型分析 BSL2 中的全部库和个体抗体特异性，并通过中和测试进行确认 与 BSL3 设施的合作者。从这些研究中，我们希望了解如何识别可能的退伍军人 患有更严重的疾病，并为开发可以成功改善病情的方法提供线索 感染严重程度并预防严重疾病。