Structure-guided vaccine design of HCV E1E2 to induce broadly neutralizing antibodies (bNAbs)

HCV E1E2 的结构引导疫苗设计可诱导广泛中和抗体 (bNAb)

• 批准号: 10409760
• 负责人: Steven Foung
• 金额: $ 38.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10409760
• 关键词: Achievement; Address; Adjuvant; Affinity; Alberta province; Animals; Antibodies; Antibody Response; Antibody titer measurement; Antigens; B-Lymphocytes; Back; Benchmarking; Binding; Bone Marrow; Cavia; Collaborations; Crystallization; Databases; Elements; Epitope Mapping; Epitopes; Foundations; Genotype; Glycoengineering; Goals; Hepatitis C Vaccine; Hepatitis C virus; Human; Immune; Immune Sera; Immunity; Immunologics; Liver; Maps; Measures; Modeling; Molecular; Molecular Conformation; Mus; Pan Genus; Plasma Cells; Plasmablast; Polysaccharides; Process; Production; Property; Protein Engineering; Proteins; Recombinants; Resistance; Resolution; Series; Structure; Study models; T cell response; T-Lymphocyte; Testing; Transmembrane Domain; Vaccinated; Vaccine Design; Vaccines; Viral; Virus; Work; base; chronic infection; design; flexibility; human monoclonal antibodies; immunogenic; immunogenicity; improved; in vivo; molecular dynamics; molecular modeling; mouse model; mutant; neutralizing antibody; polyclonal antibody; preservation; prevent; rational design; response; synergism; vaccine development

ABSTRACT - PROJECT 1 An effective HCV vaccine to prevent chronic infection will need to induce robust T and B cell responses shortly after exposure in the susceptible host, the overall goal of this U19. Project 1 focuses on a structure- guided approach to develop an immunogen that increases the neutralization potency/affinity of elicited broadly neutralizing antibodies (bNAbs) to conserved epitopes and particularly to those epitopes associated with neutralization synergy. It will be an optimized mutant (mt) E1E2 heterodimer that maintains native conformational structures for multiple bNAb epitopes to minimize viral escape. The increasing information on E2 crystal structures made possible by human monoclonal antibodies (HMAbs) that we and others have isolated and characterized as bNAbs, provides a strong foundation for the molecular approaches in rational design. A critical and complementary element is the availability of a high-resolution, functional map of both bNAb and non-neutralizing epitopes in E2 and E1E2. We have developed a large database on immunogenic domains on E2 that is derived from epitope mapping studies with a large panel of E2 bNAbs. We have also developed a large panel of bNAbs to E1E2 that will contribute to structural studies on E1E2 heterodimer in Project 4. In addition, Project 1 is built on substantial achievements with native recombinant wild-type (wt) E1E2 heterodimer as a vaccine against HCV. We have developed the necessary production and purification processes and tested to show that our wt E1E2 expresses many bNAbs epitopes and that antisera generated from vaccinated mice, guinea pigs, chimpanzees and humans contains bNAbs. However, there are gaps in the breadth of protection against some genotypes 2, 3 and 7 HCV isolates. Thus, a benchmark is to design a mt E1E2 that elicits antisera more able to neutralize these genotype isolates while maintaining a high level of neutralization towards the other genotype isolates achieved with wt E1E2. We propose to accomplish this goal by the following Specific Aims: Aim 1. Structure-guided modeling to redesign E1E2 structures to optimize presentation of E1E2 bNAb epitopes; Aim 2. Expression, purification, and characterization of modified/mutant E1E2 heterodimers; and Aim 3. Immunological characterization of against diverse HCV clades. select optimized E1E2. Finally, we aim to combine optimized E1E2 with powerful adjuvants to enhance neutralizing antibody titers (Project 3). Aim 4 is then the immunological characterization of NHP receiving an HCV vaccine optimized for bNAbs and T cell responses

摘要 - 项目1 有效的HCV疫苗可防止慢性感染，需要诱导鲁棒的T和B细胞反应 在易感宿主接触后，U19的总体目标。项目1专注于结构 - 引导的方法来开发一种增加引起的中和效力/亲和力的免疫原 对保守表位，尤其是这些表位的广泛中和抗体（BNAB） 与中和协同作用相关。它将是一种维持的优化突变体（MT）E1E2异二聚体 多种BNAB表位的天然构象结构可最大程度地减少病毒逃逸。增加 关于人类单克隆抗体（HMAB）使E2晶体结构可能成为可能的信息 其他人已经孤立并将其特征为bnab，为分子提供了坚实的基础 理性设计的方法。关键和互补的要素是高分辨率的可用性 E2和E1E2中BNAB和非中和表位的功能图。我们已经发展了一个大的 E2上免疫原性的数据库，该数据库是从表位图中得出的，该研究与大型面板 E2 bnabs。我们还为E1E2开发了一大批BNAB，这将有助于有关的结构研究 项目4中的E1E2异二聚体。此外，项目1基于本机的实质成就 重组野生型（WT）E1E2异二聚体作为针对HCV的疫苗。我们已经开发了必要的 生产和纯化过程并进行了测试以表明我们的WT E1E2表示许多BNAB 表位和由接种小鼠，豚鼠，黑猩猩和人类产生的抗血清包含 bnabs。但是，针对某些基因型2、3和7 HCV分离株的保护广度存在差距。 因此，基准是设计一种MT E1E2，该MT E1E2能够更有能力中和这些基因型分离株 同时对使用WT E1E2实现的其他基因型分离株保持高水平的中和水平。我们 建议通过以下特定目的来实现这一目标：目标1。结构引导的建模以重新设计 E1E2结构以优化E1E2 BNAB表位的呈现；目标2。表达，净化和 修饰/突变E1E2异二聚体的表征；和目标3。免疫特征 反对多样化的HCV进化枝。选择优化的E1E2。最后，我们的目标是将优化的E1E2与强大的 增强中和抗体滴度的佐剂（项目3）。 AIM 4是免疫学 接受针对BNAB和T细胞反应优化的HCV疫苗的NHP的表征