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: 10205549
• 负责人: Steven Foung
• 金额: $ 70.31万
• 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/10205549
• 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; 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; 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 预防慢性感染的有效 HCV 疫苗需要在短期内诱导强有力的 T 和 B 细胞反应 在暴露于易感宿主后，本次U19的总体目标是。项目 1 侧重于结构 - 开发免疫原的指导方法，可增加引发的中和效力/亲和力 针对保守表位，特别是针对这些表位的广泛中和抗体 (bNAb) 与中和协同作用有关。它将是一个优化的突变体 (mt) E1E2 异二聚体，可维持 多个 bNAb 表位的天然构象结构可最大限度地减少病毒逃逸。不断增加的 我们通过人单克隆抗体 (HMAb) 实现了有关 E2 晶体结构的信息 等人已分离并鉴定为 bNAb，为分子生物学奠定了坚实的基础。 理性设计的方法。一个关键且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。我们开发了大型 E2 上免疫原性结构域的数据库，该数据库源自表位作图研究，涉及大量 E2 bNAb。我们还开发了一大组针对 E1E2 的 bNAb，这将有助于结构研究 项目 4 中的 E1E2 异二聚体。此外，项目 1 是建立在原生 重组野生型 (wt) E1E2 异二聚体作为 HCV 疫苗。我们已经开发了必要的 生产和纯化过程并经过测试表明我们的 wt E1E2 表达许多 bNAb 表位以及从接种疫苗的小鼠、豚鼠、黑猩猩和人类中产生的抗血清含有 bNAb。然而，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，一个基准是设计一种 mt E1E2，它能引发更能够中和这些基因型分离株的抗血清 同时保持用wt E1E2实现的对其他基因型分离株的高水平中和。我们 建议通过以下具体目标来实现这一目标： 目标 1. 结构引导建模以进行重新设计 E1E2 结构以优化 E1E2 bNAb 表位的呈现；目标 2. 表达、纯化和 修饰/突变 E1E2 异二聚体的表征；目标 3. 免疫学特征 对抗不同的 HCV 分支。选择优化的E1E2。最后，我们的目标是将优化的 E1E2 与强大的 佐剂以增强中和抗体滴度（项目 3）。目标 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