Project 2

项目2

• 批准号: 9982207
• 负责人: Pamela J Bjorkman
• 金额: $ 58.5万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982207
• 关键词: Abbreviations; Affinity; Antibodies; Antibody titer measurement; Antibody-Dependent Enhancement; Antigen Targeting; Antigens; Binding; Cells; Collaborations; Complementarity Determining Regions; Complex; Cryoelectron Microscopy; Crystallization; Dengue; Dengue Virus; Development; Disease; E protein; Epitopes; Flavivirus; Genes; Glycoproteins; Goals; HIV-1; Human; Immunoglobulin G; Immunoglobulin Genes; Immunoglobulins; Individual; Infection; Joints; Knowledge; Laboratories; Lateral; Libraries; Light; Link; Liposomes; Masks; Membrane; Membrane Proteins; Memory B-Lymphocyte; Methods; Molecular Sieve Chromatography; Mus; Paper; Polysaccharides; Pre-Clinical Model; Pregnancy; Process; Production; Reporting; Rice; Risk; Roentgen Rays; Safety; Somatic Mutation; Structure; Surface Plasmon Resonance; Tertiary Protein Structure; Testing; Vaccination; Vaccine Antigen; Vaccines; Variant; Viral; Virion; Virus; Work; X-Ray Crystallography; Yeasts; ZIKV disease; ZIKV infection; Zika Virus; Zika virus vaccine; antigen binding; base; cross reactivity; design; experience; experimental study; fetal; global health; immunogenicity; in vivo; nanoparticle; neutralizing antibody; particle; protein E; research clinical testing; screening; virology; virus envelope

Project Summary - Project 2 (PD: Pamela Bjorkman) Zika virus (ZIKV) infection is an emerging global health concern due to the potential for fetal abnormalities when infection occurs during pregnancy. A ZIKV vaccine should preferentially elicit neutralizing antibodies (Abs) rather than non-neutralizing Abs, which may exacerbate disease caused by ZIKV or related dengue virus strains through the phenomenon of Ab-dependent enhancement (ADE). The Bjorkman lab proposes to use knowledge of the structural basis for Ab neutralization of ZIKV to design potential vaccine immunogens optimized to elicit neutralizing Abs while reducing the risk of ADE by minimizing the production of non- neutralizing Abs. The Nussenzweig, Rice, and Bjorkman laboratories have recently-established a collaboration to isolate and characterize Abs against ZIKV Envelope Domain III (ZEDIII). A group of ZIKV-infected individuals with high ZIKV neutralizing Ab activity have been found to have Abs derived from immunoglobulin genes VH3- 23/VK1-5. Several of these Abs neutralize ZIKV in vivo and are effective in challenge and in treatment experiments in mice. The structural basis for ZIKV neutralization by these Abs has been revealed by the Bjorkman lab in crystal structures of Ab–EDIII complexes. The crystal structures of these Abs bound to ZEDIII and to the counterpart domain of dengue 1 virus (DENV1) revealed a common mechanism of recognition of the ZIKV and DENV1 EDIII lateral ridge. The Bjorkman lab will extend these efforts to solve and compare structures of additional neutralizing Abs bound to their target antigens, both isolated envelope domains and virions, in order to determine which features correlate with neutralizing activity. Complexes of antigen with neutralizing Abs of varying cross-reactivity and potency (isolated by Dr. Nussenzweig and evaluated by Dr. Rice) will be crystallized and their structural features compared to identify viral vulnerabilities as well as characterize epitopes targeted by weak or non-neutralizing antibodies. The Bjorkman lab proposes to use the germline-targeting approach for ZIKV immunogen design to mitigate the potential for ADE, an approach made possible by the identification and characterization of the VH3-23/VK1-5 class of anti-ZIKV Abs. Initial immunogen design efforts will be focused on the lateral ridge epitope, which is recognized by the potent VH3-23/VK1-5 Abs. Yeast library screening methods successfully used for HIV-1 immunogen design will be adapted to identify ZEDIII variants that bind with higher affinity to iGL versions of VH3-23/VK1-5 Abs. Non-neutralizing epitopes will be masked by adding N-linked glycans or altered to reduce immunogenicity. Optimized antigens will be multimerized to generate candidate immunogens. These immunogens will be evaluated for efficacy and safety in pre-clinical models in collaboration with Drs. Nussenzweig and Rice. The goal is to develop immunogens suitable to move towards clinical testing.

项目摘要 - 项目 2（PD：Pamela Bjorkman） 由于可能导致胎儿畸形，寨卡病毒 (ZIKV) 感染已成为一个新兴的全球健康问题 当怀孕期间发生感染时，ZIKV 疫苗应优先引发中和抗体。 （抗体）而不是非中和抗体，这可能会加剧 ZIKV 或相关登革热病毒引起的疾病 Bjorkman 实验室建议使用 Ab 依赖性增强 (ADE) 现象来抑制菌株。 了解 ZIKV 抗体中和的结构基础，以设计潜在的疫苗免疫原 优化以引发中和抗体，同时通过最大限度地减少非抗体的产生来降低 ADE 的风险 中和 Abs。 Nussenzweig、Rice 和 Bjorkman 实验室最近建立了合作关系，以分离 并表征针对 ZIKV 包膜结构域 III (ZEDIII) 的一组 ZIKV 感染个体。 已发现具有高 ZIKV 中和抗体活性的抗体源自免疫球蛋白基因 VH3- 23/VK1-5。其中几种抗体可在体内中和 ZIKV，并且在攻击和治疗中有效。 小鼠实验揭示了这些抗体中和 ZIKV 的结构基础。 Bjorkman 实验室研究 Ab-EDIII 复合物的晶体结构 这些 Ab 与 ZEDIII 结合的晶体结构。 登革热 1 病毒 (DENV1) 的对应结构域揭示了一种常见的识别机制 Bjorkman 实验室将扩展这些努力来解决和比较 ZIKV 和 DENV1 EDIII 侧脊。 与其包膜靶抗原结合的额外中和抗体的结构，包括分离的结构域和 病毒粒子，以确定哪些特征与抗原复合物的中和活性相关。 不同交叉反应性和效力的中和抗体（由 Nussenzweig 博士分离并由 Nussenzweig 博士评估） 大米）将被结晶并与它们的结构特征进行比较，以识别病毒的脆弱性以及 表征弱或非中和抗体靶向的表位。 Bjorkman 实验室建议使用种系靶向方法进行 ZIKV 免疫原设计 减轻 ADE 的潜力，这种方法通过识别和表征 VH3-23/VK1-5 类抗 ZIKV 抗体的初始免疫原设计工作将集中在侧脊。 表位，可被有效的 VH3-23/VK1-5 酵母文库筛选方法成功识别。 用于 HIV-1 免疫原设计的 ZEDIII 变体将适用于识别与 iGL 具有更高亲和力的 ZEDIII 变体 VH3-23/VK1-5 Abs 的版本将通过添加 N 连接聚糖或改变来掩盖。 以降低免疫原性。优化的抗原将被多聚化以产生候选免疫原。 将与 Drs 合作在临床前模型中评估免疫原的有效性和安全性。 Nussenzweig 和 Rice 的目标是开发适合进行临床测试的免疫原。