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: 10490889
• 负责人: Wayne A. Marasco
• 金额: $ 87.69万
• 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/10490889
• 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; Childhood; China; Clinical; Coronavirus; Coronavirus Infections; Data; Databases; Development; Disease; 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; Lead; Literature; Malignant Childhood Neoplasm; Memory B-Lymphocyte; Middle East Respiratory Syndrome; Modeling; Molecular; Monoclonal Antibodies; Mus; Patients; Peptides; Persons; Plasma; Plasma Cells; 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; Sampling; Serology; Severe Acute Respiratory Syndrome; Specificity; Stains; T-Cell Activation; Techniques; Testing; Vaccine Design; Virus; 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; 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.

项目摘要 2019年，中国武汉的新型人类贝塔科罗纳病毒SARS-COV2的出现迅速发展 进入全球大流行。超过1亿人被感染了，已经有数百万 死亡人数。从无症状的情况下，covid-19疾病呈现的方式也存在很大的差异 感染致死。与成年人相比，儿童的Covid-19疾病总体上更为轻度或无症状。一 可能解释这种差异的假设是，由于经常出现，儿童与SARS-COV2具有跨免疫性 早期暴露于引起温和呼吸系统疾病的全球循环冠状病毒（HCOV）。 流行的HCOV和SARS-COV2是否有一定程度的跨免疫力 成年，可以提供一定程度的保护与Covid-19疾病的保护是该R01应用的主题。 我们的主要目的是提供抗HCOV/SARS-COV2尖峰的血清学和分子证据 可反应性和中和抗体，可以在体内提供保护SARS-COV2。我们有一个IRB- 批准的方案以收集250 COVID-19个人的血液样本。我们的共同队列由5个组成 包括成人和儿科癌症患者，成人和小儿医疗保健提供者和成人的组 没有共同患者接触。此外，我们还将研究我们的大大季节性季节性流感队列 预先存在的抗SARS-COV2 S ABS的证据。在AIM 1中，我们将量化抗S HCOV抗体的当下 并量化其对SARS-COV2S的交叉反应性。 S表达S子域的细胞和ELISA用于表位图。在Subaim 1B中，选定的血浆样品 在每个研究组中，将使用将被转移的等离子IgG的亲和力柱纯化 并从4个HCOV或SARS-COV2尖峰柱之一洗脱，并测试了交叉结合，交叉FC效应器 活性和跨中和活性。在Subaim 1C中，这些纯化的IgG将在HACE2小鼠的体内进行测试 针对SARS-COV2挑战的交叉保护。在AIM 2中，我们将建立分子基础 COV中的方向免疫力可以通过共同提供对HCOV和SARS-COV2的交叉免疫力 尖峰表位识别。在Subaim 2a中，我们将执行记忆B（MB）细胞筛选，以实现S交叉的存在 结合。 facs将分离至少一个HCOV的蛋白和SARS-COV2 S的单个MB细胞，它们 同源VH/VL基因克隆，以整个IgG1 mAb表示，并测试了交叉结合，病毒中和 和FC效应子活性针对不同的Hocovs，SARS和SARS-COV2。在subiaim 2b中，mabs cross- COV活动将在小鼠和仓鼠模型中进行测试，以防止SARS-COV2挑战。在 Subaim 2C，我们将调整新型Libraseq技术以捕获单个或多尖峰结合特异性， 选定BM细胞的BCR曲目和转录组，研究潜在的不同进化起源 单尖峰和多尖峰结合细胞之间可能存在。 R01赠款将提供原理证明 HCOV/SARS-COV2 AB跨免疫的分子研究可能有助于Covid-19疫苗设计。