Structural and mechanistic insights into antibody neutralization of human metapneumovirus

人类偏肺病毒抗体中和的结构和机制见解

基本信息

批准号：
10654625
负责人：
Jarrod Mousa
金额：
$ 5.35万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10654625
关键词：
Acute Address Adult Affinity Amino Acids Animal Model Antibodies Antibody Specificity Antigens B-Lymphocytes Binding Blood specimen Cells Characteristics Child Chimeric Proteins Chronic Obstructive Pulmonary Disease Circulation Clinic Collaborations Complex Cotton Rats Data Disease Electron Microscopy Epidemiologist Epitopes Genes Genetic Goals Growth Human Human Metapneumovirus Hybridomas Immune response Immunity Immunocompromised Host Immunologics Immunologist Inbred BALB C Mice Infection Mediating Memory B-Lymphocyte Modeling Molecular Molecular Conformation Monoclonal Antibodies Multiple Myeloma Mus Mutagenesis Mutation National Institute of Allergy and Infectious Disease Negative Staining Nicaragua Nicaraguan Prevention Process Proteins Research Personnel Research Project Grants Respiratory Disease Respiratory Tract Infections Role Site Somatic Mutation Specificity Structure Subgroup Surveillance Modeling Technology Testing Therapeutic Therapeutic Uses Treatment Efficacy Vaccine Design Vaccines Viral Virus X-Ray Crystallography cohort design disorder prevention epidemiologic data human monoclonal antibodies human pathogen human subject in vivo infectious disease treatment insight neutralizing antibody neutralizing monoclonal antibodies novel therapeutics permissiveness prevent rational design response scaffold structural biology three dimensional structure vaccine strategy

项目摘要

Human metapneumovirus (hMPV) is a leading cause of acute respiratory tract infection in children, and is being recognized more in recent years as a significant cause of disease in the immunocompromised, and in exacerbations of chronic obstruction pulmonary disease in adults. Although hMPV was discovered nearly two decades ago, no vaccine or therapeutic is currently available for prevention or treatment of hMPV disease, and we have a poor understanding of protective immune responses generated in response to natural infection. The sole target of neutralizing antibodies is the hMPV fusion (F) protein. The overall objective of this research project grant is to determine the human humoral immune response to the hMPV F protein. The data generated from this research project addresses our overall hypothesis that potent neutralizing antibody activity against hMPV is a result of (1) extensive antibody somatic mutation, and of (2) recognition of specific epitopes on the F protein that inhibit structural transitions critical for the viral attachment and fusion processes. Utilizing blood samples from human subjects, human memory B cells will be screened for antibody-specificity targeting the pre- fusion and post-fusion forms of the hMPV F protein, at which point the B cells will be fused to myeloma cells to generate human hybridomas. Isolated human mAbs targeting the hMPV F protein will be characterized to determine affinity, fine epitope specificity, neutralization potency, and for select mAbs we will determine the three-dimensional structure in complex with the hMPV F protein. These data will inform rational vaccine design of an effective hMPV vaccine. Determining the major neutralizing epitopes on the hMPV F protein will inform vaccine design by identifying amino acid regions important for hMPV F immunity, which can be incorporated into subunit or scaffolded vaccines to elicit target antibodies. Additionally, we will examine if these mAbs can prevent hMPV disease in vivo, which will be a prelude to use in the clinic. In Specific Aim 1, we will determine the ontogeny, neutralizing mechanisms, subgroup-reactivity, and conformational specificity of several hMPV F- specific human mAbs in both adults and children. In Specific Aim 2, we will determine the structural correlates of potent antibody neutralization of hMPV. In Specific Aim 3, we will determine the therapeutic efficacy of our top candidate human mAbs for prevention and treatment of hMPV disease in both mice and cotton rat models of infection.

人类偏肺病毒（hMPV）是儿童急性呼吸道感染的主要原因，目前正在研究中近年来，人们越来越认识到这是免疫功能低下者患病的一个重要原因，成人慢性阻塞性肺病的恶化。尽管 hMPV 被发现近两几十年前，目前还没有疫苗或治疗方法可用于预防或治疗 hMPV 疾病，并且我们对自然感染产生的保护性免疫反应知之甚少。这中和抗体的唯一目标是 hMPV 融合 (F) 蛋白。本研究的总体目标项目资助的目的是确定人类对 hMPV F 蛋白的体液免疫反应。生成的数据这个研究项目提出了我们的总体假设，即有效的中和抗体活性 hMPV 是 (1) 广泛的抗体体细胞突变和 (2) F 上特定表位识别的结果抑制对病毒附着和融合过程至关重要的结构转变的蛋白质。利用血液从人类受试者的样本中，将筛选人类记忆 B 细胞的抗体特异性，该抗体特异性针对前 hMPV F 蛋白的融合和融合后形式，此时 B 细胞将与骨髓瘤细胞融合产生人类杂交瘤。靶向 hMPV F 蛋白的分离人单克隆抗体将被表征为确定亲和力、精细表位特异性、中和效力，对于选定的单克隆抗体，我们将确定与 hMPV F 蛋白复合的三维结构。这些数据将为合理的疫苗设计提供信息有效的 hMPV 疫苗。确定 hMPV F 蛋白上的主要中和表位将提供信息通过识别对 hMPV F 免疫重要的氨基酸区域来设计疫苗，可以将其纳入亚单位或支架疫苗以引发目标抗体。此外，我们将检查这些单克隆抗体是否可以预防 hMPV在体内的疾病研究，这将是其应用于临床的前奏。在具体目标 1 中，我们将确定几种 hMPV F-的个体发育、中和机制、亚群反应性和构象特异性成人和儿童的特定人类单克隆抗体。在具体目标 2 中，我们将确定结构相关性 hMPV 的有效抗体中和。在具体目标 3 中，我们将确定我们的治疗效果用于预防和治疗小鼠和棉鼠模型中 hMPV 疾病的最佳候选人单克隆抗体的感染。