B-Cell Epitope Discovery and Mechanisms of Antibody Protection for ZIKV, MARV, and VEEV Envelope Proteins

ZIKV、MARV 和 VEEV 包膜蛋白的 B 细胞表位发现和抗体保护机制

• 批准号: 10021078
• 负责人: BENJAMIN DORANZ
• 金额: $ 136.33万
• 依托单位: INTEGRAL MOLECULAR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-16 至 2020-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10021078
• 关键词: Alanine; Antibodies; Antibody-Dependent Enhancement; Antibody-mediated protection; B-Lymphocyte Epitopes; B-Lymphocytes; Binding Sites; Biochemical; Cells; Clinical; Clone Cells; Computer software; Contractor; Cryoelectron Microscopy; Data; Databases; Epitope Mapping; Epitopes; Evaluation; Family member; Frankfurt-Marburg Syndrome Virus; Human; Immune; Immune response; In Vitro; Individual; Infection; Maps; Measures; Molecular Conformation; Monoclonal Antibodies; Mutagenesis; Pathogenesis; Process; Resources; Role; Sampling; Scanning; Shotguns; Structure; Support Contracts; Testing; Vaccinated; Validation; Venezuelan Equine Encephalitis Virus; Viral Pathogenesis; Virus; Zika Virus; biophysical properties; env Gene Products; human subject; in vivo; next generation sequencing; pathogenic virus; protein complex

This contract supports the identification of human B cell epitopes derived from three viral pathogens; Zika, Marburg and Venezuelan equine encephalitis viruses, combined with basic studies to understand protective immunity mediated by antibodies. Milestones include 1) epitope identification; 2) epitope validation that must include in vitro evaluation using human samples; 3) submission of epitope data, computer software, and structural data to the Immune Epitope Database and Analysis Resource; and 4) studies to help understand the mechanisms of protection or pathogenesis elicited by antibodies associated with the newly identified epitopes. The Contractor will discover and characterize B cell epitopes on the Envelope (Env) proteins of Zika virus (ZIKV prM/E), Marburg virus (MARV GP), and Venezuelan equine encephalitis virus (VEEV E1/E2). They will generate new monoclonal antibodies (MAbs) by B cell cloning and next-generation sequencing using human clinical samples from both naturally infected and vaccinated individuals; Map the binding site of each Mab; Validate the functional relevance of each epitope to neutralize or, in the case of ZIKV MAbs, to enhance infectivity; and determine the mechanism of action of each Mab.

该合同支持鉴定来自三种病毒病原体的人类B细胞表位;寨卡病毒，马尔堡病毒和委内瑞拉马脑炎病毒，并结合基础研究以了解抗体介导的保护性免疫。重要性包括1）表位鉴定; 2）表位验证，其必须包括使用人样品的体外评价; 3）向免疫表位数据库和分析资源提交表位数据、计算机软件和结构数据;以及4）研究以帮助理解与新鉴定的表位相关的抗体引起的保护或发病机制。承包商将发现并表征寨卡病毒（ZIKV prM/E）、马尔堡病毒（MARV GP）和委内瑞拉马脑炎病毒（VEEV E1/E2）包膜（Env）蛋白上的B细胞表位。他们将通过B细胞克隆和下一代测序，使用来自自然感染和接种疫苗个体的人类临床样本生成新的单克隆抗体（MAb）;绘制每个MaB b的结合位点;确定每个表位的功能相关性，以中和或在ZIKV MAb的情况下增强感染性;并确定每个MaB b的作用机制。