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

• 批准号: 10439797
• 负责人: Jarrod Mousa
• 金额: $ 55.48万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10439797
• 关键词: Acute; Address; Adult; Affinity; Amino Acids; Animal Model; Antibodies; Antibody Specificity; Antigens; B-Lymphocytes; Binding; Blood Circulation; Blood specimen; Cells; Characteristics; Child; Chimeric Proteins; Chronic Obstructive Pulmonary Disease; Clinic; Collaborations; Complex; Cotton Rats; Crystallization; Data; Disease; Electron Microscopy; Epidemiologist; Epitopes; Genes; 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; Molecular Genetics; 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; base; 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; prevent; rational design; response; scaffold; structural biology; three dimensional structure; vaccine strategy

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.

人元病毒（HMPV）是儿童急性呼吸道感染的主要原因，并且正在 近年来，人们更多地认为是免疫功能低下的重要原因，在 成人慢性阻塞肺疾病的加剧。虽然发现了HMPV将近两个 几十年前，目前没有疫苗或治疗性可用于预防或治疗HMPV疾病，并且 我们对响应自然感染产生的保护性免疫反应的理解很差。这 中和抗体的唯一靶标是HMPV融合（F）蛋白。这项研究的总体目标 项目授予是为了确定人类对HMPV F蛋白的免疫力反应。生成的数据 从这项研究项目解决了我们的总体假设，即对抗体活性有效 HMPV是（1）广泛的抗体体细胞突变的结果，以及（2）识别F上的特定表位 抑制结构过渡的蛋白质对病毒附着和融合过程至关重要。利用血液 来自人类受试者的样本，将筛选人类记忆B细胞的抗体特异性，以靶向前 HMPV F蛋白的融合和融合形式，此时B细胞将融合到骨髓瘤细胞中 产生人类杂交瘤。靶向HMPV F蛋白的分离的人mAb将被表征为 确定亲和力，精细的表位特异性，中和效力，对于精选的mAb，我们将确定 与HMPV F蛋白复合物中的三维结构。这些数据将为理性疫苗设计提供信息 有效的HMPV疫苗。确定HMPV F蛋白上的主要中和表位将告知 通过识别对HMPV F免疫重要的氨基酸区域来设计疫苗的设计，可以将其纳入 亚基或脚手架疫苗引起靶抗体。此外，我们将检查这些mab是否可以防止 体内HMPV疾病将是诊所使用的前奏。在特定目标1中，我们将确定 多种HMPV F-的本体发育，中和机制，亚组反应性和构象特异性 成人和儿童的特定人物单位。在特定的目标2中，我们将确定结构相关性 HMPV的有效抗体中和。在特定目标3中，我们将确定我们的治疗功效 在小鼠和棉花大鼠模型中，用于预防和治疗HMPV疾病的顶级人物单元 感染。