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

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。 使用Integral Molecular的高通量表位作图平台（表达在每个氨基酸位置具有点突变的S蛋白的细胞阵列）快速作图特异性抗体。

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).