Serologic and Molecular Studies of human anti-hCoV antibody cross-immunity and protective responses among endemic HCoVs and SARS-CoV2

人类抗 hCoV 抗体交叉免疫和地方性 HCoV 与 SARS-CoV2 之间保护性反应的血清学和分子研究

• 批准号: 10689125
• 负责人: Wayne A. Marasco
• 金额: $ 87.47万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-17 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10689125
• 关键词: 2019-nCoV; ACE2; Address; Adult; Affinity; Antibodies; Antibody Response; Antibody-mediated protection; B-Lymphocytes; Binding; Biological Assay; Blood specimen; CD8-Positive T-Lymphocytes; COVID-19; COVID-19 patient; COVID-19 vaccine; Cancer Patient; Cells; Cessation of life; Child; China; Clinical; Coronavirus; Coronavirus Infections; Data; Databases; Development; Disease; Disparity; Enzyme-Linked Immunosorbent Assay; Epidemic; Epitope Mapping; Epitopes; Exposure to; Extracellular Domain; Generations; Goals; Grant; Hamsters; Health Personnel; Human; IgG1; Immune system; Immunity; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Inflammatory; Institutional Review Boards; Knowledge; Literature; Malignant Childhood Neoplasm; Memory B-Lymphocyte; Middle East Respiratory Syndrome; Modeling; Molecular; Monoclonal Antibodies; Mus; Patients; Peptides; Persons; Plasma; Population; Principal Investigator; Property; Prophylactic treatment; Proteins; Protocols documentation; Recording of previous events; Research; Risk Factors; SARS coronavirus; SARS-CoV-2 antibody; SARS-CoV-2 exposure; SARS-CoV-2 immunity; SARS-CoV-2 infection; SARS-CoV-2 spike protein; Sampling; Seasons; Serology; Severe Acute Respiratory Syndrome; Specificity; Stains; T-Cell Activation; Techniques; Testing; Vaccine Design; Virus; Writing; betacoronavirus; cohort; comorbidity; convalescent plasma; coronavirus disease; cross immunity; cross reactivity; early onset; gene cloning; human coronavirus; immunogenic; in vivo; in vivo evaluation; influenza virus vaccine; neutralizing antibody; novel; pandemic disease; pediatrician; pre-pandemic; receptor; receptor binding; respiratory; response; screening; seasonal influenza; transcriptome

Project Summary The emergence of the novel human betacoronavirus SARS-CoV2 in Wuhan, China in 2019 has rapidly evolved into a worldwide pandemic. Over a 100 million people have been infected and there have been several million deaths. There is also great disparity in the manner in which COVID-19 illness presents, from asymptomatic infection to death. COVID-19 illness in children is overall more mild or asymptomatic compared to adults. One hypothesis that may explain this disparity is that children have cross-immunity to SARS-CoV2 due to frequent early exposure to globally circulating human coronaviruses (HCoVs) that cause a milder respiratory illness. Whether there is some level of cross-immunity between the endemic HCoVs and SARS-CoV2 that carries into adulthood and can provide some level of protection from COVID-19 disease is the subject of this R01 application. Our primary goal is to provide serologic and molecular evidence of anti-HCoV/SARS-CoV2 spike (S) cross- reactive and neutralizing antibodies that can provide protection against SARS-CoV2 in vivo. We have an IRB- approved protocol to collect blood samples on 250 COVID-19 individuals. Our COVID cohort is comprised of 5 groups that includes adult and pediatric cancer patients, adult and pediatric healthcare providers and adults without COVID patient contact. In addition, we will study our pre-pandemic seasonal influenza cohort for evidence of pre-existing anti-SARS-CoV2 S Abs. In Aim 1 we will quantify the present of anti-S HCoV antibodies and quantitate their cross-reactivity to SARS-CoV2 S. The studies in subaim 1A will include FACS staining of S expressing cells and ELISAs of S subdomains for epitope mapping. In subaim 1B, selected plasma samples within each study group will be used for affinity column purification of plasma IgGs that will be passaged over and eluted from one of 4 HCoV or SARS-CoV2 spike columns and tested for cross-binding, cross-Fc effector activity and cross-neutralization activity. In subaim 1C, these purified IgGs will be tested in vivo in hACE2 mice for cross-protection against SARS-CoV2 challenge. In Aim 2 we will establish the molecular basis by which bi- directional immunity to among CoVs could provide cross immunity to HCoVs and SARS-CoV2 through common spike epitope recognition. In subaim 2A, we will perform memory B (mB) cell screening for presence of S cross- binding. Single mB cells that bind at least one hCoV S protein and SARS-CoV2 S will be isolated by FACS, their cognate VH/VL genes cloned, expressed as whole IgG1 mAbs and tested for cross-binding, virus neutralization and Fc effector activity against the different HoCoVs, SARS and SARS-CoV2. In subaim 2B, mAbs with cross- CoV activity will be tested in mouse and hamster models for protection against SARS-CoV2 challenge. In subaim 2C, we will adapt the novel LibraSeq technique to capture the single or multi-spike binding specificity, BCR repertoires and transcriptomes of selected Bm cells to study the potential different evolutionary origins that may exist between mono-spike and multi-spike binding cells. This R01 grant will provide proof-of-principle molecular studies of HCoV/SARS-CoV2 Ab cross-immunity that may aid in COVID-19 vaccine design.

项目概要 2019 年在中国武汉出现的新型人类乙型冠状病毒 SARS-CoV2 迅速演变 陷入全球性大流行。超过1亿人被感染，已有数百万人被感染 死亡人数。 COVID-19 疾病的表现方式也存在很大差异，从无症状到 感染致死。与成人相比，儿童的 COVID-19 疾病总体上更为轻微或无症状。一 可以解释这种差异的假设是，由于频繁接触儿童，儿童对 SARS-CoV2 具有交叉免疫力。 早期接触全球流行的人类冠状病毒（HCoV），会导致较轻微的呼吸道疾病。 地方性 HCoV 和 SARS-CoV2 之间是否存在一定程度的交叉免疫，从而传播病毒 R01 申请的主题是成年期并能提供一定程度的针对 COVID-19 疾病的保护。 我们的主要目标是提供抗 HCoV/SARS-CoV2 刺突 (S) 交叉的血清学和分子证据 反应性和中和性抗体，可以在体内提供针对 SARS-CoV2 的保护。我们有一个IRB- 已批准收集 250 名 COVID-19 个体血液样本的方案。我们的新冠肺炎队列由 5 名 包括成人和儿童癌症患者、成人和儿童医疗保健提供者以及成年人的群体 无需接触新冠肺炎患者。此外，我们将研究大流行前的季节性流感队列 预先存在的抗 SARS-CoV2 S 抗体的证据。在目标 1 中，我们将量化抗 S HCoV 抗体的存在 并量化它们与 SARS-CoV2 S 的交叉反应性。 subaim 1A 中的研究将包括 FACS 染色 S 表达细胞和 S 子域的 ELISA，用于表位作图。在子目标 1B 中，选定的血浆样本 每个研究组内将用于亲和柱纯化将传代的血浆 IgG 从 4 个 HCoV 或 SARS-CoV2 尖峰柱之一中洗脱，并测试交叉结合、交叉 Fc 效应子 活性和交叉中和活性。在 subaim 1C 中，这些纯化的 IgG 将在 hACE2 小鼠体内进行体内测试 针对 SARS-CoV2 挑战的交叉保护。在目标 2 中，我们将建立双- 对 CoV 之间的定向免疫可以通过共同作用为 HCoV 和 SARS-CoV2 提供交叉免疫 刺突表位识别。在 subaim 2A 中，我们将进行记忆 B (mB) 细胞筛选，以确定是否存在 S 交叉 绑定。将通过 FACS 分离结合至少一种 hCoV S 蛋白和 SARS-CoV2 S 的单个 mB 细胞，其 克隆同源 VH/VL 基因，表达为完整 IgG1 mAb，并测试交叉结合、病毒中和 和 Fc 效应子活性，针对不同的 HoCoV、SARS 和 SARS-CoV2。在 subaim 2B 中，单克隆抗体具有交叉 冠状病毒活性将在小鼠和仓鼠模型中进行测试，以防止 SARS-CoV2 的攻击。在 subaim 2C，我们将采用新颖的 LibraSeq 技术来捕获单或多尖峰结合特异性， 选定 Bm 细胞的 BCR 库和转录组，用于研究潜在的不同进化起源 可能存在于单刺和多刺结合细胞之间。此 R01 拨款将提供原理证明 HCoV/SARS-CoV2 Ab 交叉免疫的分子研究可能有助于 COVID-19 疫苗的设计。