The evolution of dengue virus-reactive circulating antibody repertoire

登革热病毒反应性循环抗体库的进化

• 批准号: 10647572
• 负责人: Eva Harris
• 金额: $ 24.99万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647572
• 关键词: Acute; Address; Antibodies; Antibody Repertoire; Antibody Response; Antibody-Dependent Enhancement; Antibody-mediated protection; B cell repertoire; B-Lymphocytes; B-cell receptor repertoire sequencing; Biological Assay; Blood Vessels; Cell Compartmentation; Cellular biology; Child; Childhood; Circulation; Cohort Studies; Communities; Complex; Convalescence; Coupled; Dengue; Dengue Infection; Dengue Vaccine; Dengue Virus; Disease; Epidemiology; Evolution; Flavivirus; Frequencies; Future; Generations; Genomics; Human; Humoral Immunities; Immune; Immune response; Immunity; Immunoglobulin G; Immunologics; Immunology; Individual; Infection; Investigation; Licensing; Maintenance; Memory B-Lymphocyte; Monoclonal Antibodies; Neutralization Tests; Nicaragua; Persons; Plasma; Plasma Cells; Population; Populations at Risk; Predisposition; Primary Infection; Prospective, cohort study; Proteomics; Recombinants; Research; Resolution; Risk; Sampling; Serology; Serotyping; Shapes; Sorting; Specificity; Standardization; System; Techniques; Technology; Time; Vaccine Design; Vaccinee; Vaccines; Viral; Viral Antigens; Virus; Virus Diseases; bioinformatics pipeline; climate change; convalescent plasma; defined contribution; human disease; improved; infection risk; innovation; mosquito-borne; multidisciplinary; neutralizing antibody; novel; recruit; response; secondary infection; sequencing platform; stem; vaccine strategy; virology

ABSTRACT The four dengue virus serotypes (DENV1-4) cause the most important mosquito-borne viral disease of humans, with ~50 million dengue cases annually and over 3 billion people worldwide at risk of infection. Yet, no treatment is currently approved for use, and the only registered vaccine is not safe and effective in all populations. The overall approach of this study is to take advantage of unique longitudinal sample sets from our long-standing cohort study in Nicaragua to address complex questions about DENV antibody and B cell immunology in a relevant epidemiological context. While antibodies generated in primary DENV infection confer protection against the same serotype, they can either protect against or enhance a subsequent infection with a different DENV serotype; however, after a secondary DENV infection, individuals acquire broader protection against multiple DENV serotypes, and the risk of severe disease is lowered. There is a critical gap in understanding the generation and maintenance of broadly neutralizing circulating antibodies and B cell immunity stemming from secondary DENV infection. Our preliminary results show that expansion of B cell recall contributes to acute secondary DENV immunity and that post-secondary memory B cells (MBCs) display broader cross- serotype neutralization as compared to post-primary MBCs. We hypothesize that the recruitment of B cell memory from a primary DENV infection into acute secondary circulating IgG, and then substantial addition of new lineages from the secondary DENV infection into long-lasting serological and B cell repertoires, together shape the distinct broadly protective profile of durable post-secondary dengue immunity. In this study, we will investigate how pre-existing MBCs generated during primary DENV infection and newly elicited lineages in secondary DENV infection contribute to broad neutralization. In particular, we will identify whether broadly neutralizing antibodies originate from the pre-existing B cell compartment or are newly elicited in secondary infection (Aim 1), and how they are maintained in serological or B cell repertoires for future protection (Aim 2). We will use innovative viral antigens for isolation of DENV-specific antibodies and B cells coupled with state-of- the-art IgG antibody proteomics (Ig-Seq) and B cell receptor sequencing (BCR-Seq) platforms, with robust bioinformatics pipelines. Selected representative monoclonal antibodies will be expressed and functionally evaluated using established, standardized neutralization assays. We have assembled a team with complementary expertise in B cell biology, flavivirus virology, immunology, and epidemiology, coupled with cutting-edge ‘antibodyomic’ serological and genomic sequencing technologies and analytic systems. Overall, the proposed research will begin to define at the cellular and serological level the evolution of antibody- mediated immunity that generates broad neutralization against DENV upon secondary infection in order to guide improved vaccine design strategies.

抽象的 四种登革热病毒血清型（DENV1-4）引起人类最重要的蚊媒病毒性疾病， 每年约有 5000 万登革热病例，全世界有超过 30 亿人面临感染风险，但尚无治疗方法。 目前已批准使用，唯一注册的疫苗并不对所有人群安全有效。 这项研究的总体方法是利用我们长期积累的独特的纵向样本集 尼加拉瓜的队列研究旨在解决有关 DENV 抗体和 B 细胞免疫学的复杂问题 而原发性 DENV 感染中产生的抗体可提供针对相关流行病学的保护。 相同血清型，它们可以预防或增强不同 DENV 的后续感染 然而，继发性 DENV 感染后，个体获得了更广泛的针对多种病毒的保护。 DENV 血清型和严重疾病的风险降低了。 广泛中和循环抗体的产生和维持以及 B 细胞免疫干预 我们的初步结果表明，B 细胞回忆的扩展有助于预防继发性 DENV 感染。 急性继发性 DENV 免疫和二次记忆 B 细胞 (MBC) 表现出更广泛的交叉免疫 与原代后 MBC 相比，我们在 B 细胞的招募方面遇到了困难。 将原发性 DENV 感染记忆转化为急性继发性循环 IgG，然后大量添加 从继发性 DENV 感染到持久的血清学和 B 细胞库的新谱系 在这项研究中，我们将塑造持久的登革热免疫的独特广泛保护特征。 研究在原发性 DENV 感染期间如何产生预先存在的 MBC 以及如何在 继发性 DENV 感染有助于广泛中和，特别是，我们将确定是否广泛中和。 中和抗体源自预先存在的 B 细胞区室或在次级细胞中新引发 感染（目标 1），以及如何将它们保留在血清学或 B 细胞库中以供未来保护（目标 2）。 我们将使用创新的病毒抗原来分离 DENV 特异性抗体和 B 细胞，并结合状态- 最先进的 IgG 抗体蛋白质组学 (Ig-Seq) 和 B 细胞受体测序 (BCR-Seq) 平台，具有强大的功能 选定的代表性单克隆抗体将被表达并发挥功能。 我们已经组建了一个团队，使用已建立的标准化中和测定进行评估。 B 细胞生物学、黄病毒病毒学、免疫学和流行病学方面的互补专业知识，加上 尖端的“抗体组学”血清学和基因组测序技术和分析系统。 拟议的研究将开始在细胞和血清学水平上定义抗体的进化 介导的免疫，在继发感染时产生针对 DENV 的广泛中和作用 指导改进的疫苗设计策略。