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

ZIKV、MARV、VEEV 包膜蛋白的 B 细胞表位发现和抗体保护机制

• 批准号: 10668917
• 负责人: BENJAMIN DORANZ
• 金额: $ 104.46万
• 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 2023-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10668917
• 关键词: Alanine; Amino Acids; 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; Human; Immune; Immune response; In Vitro; Infection; Maps; Marburgvirus; Measures; Molecular; Molecular Conformation; Monoclonal Antibodies; Mutagenesis; Pathogenesis; Point Mutation; Positioning Attribute; Process; Proteins; Resources; Role; Sampling; Scanning; Shotguns; Support Contracts; Testing; Vaccinated; Vaccinee; Validation; Venezuelan Equine Encephalitis Virus; Viral Pathogenesis; Virus; ZIKA; 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. Rapid mapping of specific antibodies using integral molecular' s high throughput epitope mapping platform (arrays of cells expressing S protein with point mutations at every amino acid position).

该合同支持识别来自三种病毒病原体(寨卡病毒、马尔堡病毒和委内瑞拉马脑炎病毒)的人类B细胞表位，并结合基础研究了解抗体介导的保护性免疫。里程碑包括1)表位识别；2)表位验证，必须包括使用人类样本进行体外评估；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)帮助理解与新识别的表位相关的抗体所引发的保护或致病机制的研究。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)包膜蛋白(Env)上的B细胞表位。他们将利用来自自然感染和接种疫苗的人的临床样本，通过B细胞克隆和下一代测序产生新的单抗；绘制每一单抗的结合位置；验证每一表位的功能相关性，以中和或在ZIKV单抗的情况下，增强传染性；并确定每一单抗的作用机制。 使用积分分子的S高通量表位作图平台(表达S蛋白的细胞阵列，每个氨基酸位置都有点突变)快速绘制特异性抗体图谱。