Novel HCV vaccine antigens targeting conserved neutralizing epitopes

针对保守中和表位的新型 HCV 疫苗抗原

基本信息

批准号：
9096641
负责人：
Jiang Zhu
金额：
$ 28.97万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9096641
关键词：
Animal Model Animals Antibodies Antibody Repertoire Antibody Response Antigen Targeting Antigens B cell repertoire B-Lymphocytes Bacteriophages Biological Assay Cell Lineage Cell Separation Chimeric Proteins Chronic Collaborations Complex Data Development Engineering Epitope Mapping Epitopes Ferritin Flaviviridae Flow Cytometry Genes Genetic Genotype Glycoproteins HIV-1 HIV-1 vaccine Hepatitis C Hepatitis C Antibodies Hepatitis C Vaccination Hepatitis C Vaccine Hepatitis C virus Human Immunization Immunoglobulin Somatic Hypermutation Individual Infection Laws Length Link Maps Microfluidics Modeling Molecular Conformation Mus Mutation Nucleosome Core Particle Peptides Pharmaceutical Preparations Polysaccharides Population Primates Property Protein Engineering Public Health Reporting Research Respiratory Syncytial Virus Vaccines Role Sampling Scaffolding Protein Series Serum Structure Subunit Vaccines Surface Technology Testing Time Transcript Treatment Cost Vaccination Vaccine Antigen Vaccine Design Vaccine Research Vaccines Variant Viral Antigens Virus Virus-like particle base design env Gene Products experience hepatitis C virus envelope 2 protein immunogenic immunogenicity innovation iterative design nanoparticle neutralizing antibody next generation sequencing nonhuman primate novel particle prophylactic research study response scaffold small molecule structural biology success trend vaccine candidate vaccine development vaccine trial virology

项目摘要

Project Summary It has been estimated that ~3% of the word population is infected with hepatitis C virus (HCV). Recent studies have identified a number of neutralizing epitopes on the HCV E1 and E2 glycoproteins and demonstrated with atomic structures how broadly neutralizing antibodies (bnAbs) interact with these epitopes. The 2.65Å crystal structure of HCV E2 core (E2c) in complex with a bnAb (AR3C) revealed atomic details of this long-sought target, providing a structural basis for rational design of HCV vaccines. In Project 2 of this U19 proposal, we will combine the latest findings in HCV structural biology and cutting-edge technologies in computational protein design and next-generation sequencing (NGS) of B-cell repertoire to facilitate HCV immunogen design for the induction of bnAbs in vaccination. The specific aims are: (1) to develop epitope-focused immunogens and E2 core-based immunogens. Crystal structures of E2 and E1 neutralizing epitopes in complex with bnAbs have been determined recently. We hypothesize that heterologous protein scaffolds presenting a grafted HCV epitope can induce cross-neutralizing antibodies to the epitope. We also hypothesize that the E2c domain can be optimized as a subunit vaccine immunogen to induce cross-neutralizing antibodies to the conserved epitopes presented on the E2 surface. Based on the crystal structure of E2c, we will shorten the variable loops, engineer the surface N-linked glycans, and introduce space-filling mutations to the front layer region to stabilize the E2c conformation. Successfully designed and expressed HCV antigens will be displayed multivalently on ferritin particle and bacteriophage Qβ VLP. (2) to use NGS repertoire profiling to assess antibody response in mouse and non-human primate immunization. Promising immunogen candidates will be first tested in mice. In collaboration with the Law lab, an array of HCV-specific immunological assays will be available and performed to map the serum antibody responses. A small set of designed immunogens that can induce cross-neutralizing antibodies in mice will be tested in non-human primates (NHPs) at the Southwest National Primate Research Center (SNPRC). Flow cytometry-based single B-cell sorting and microfluidics-based B-cell encapsulation will be utilized to isolate HCV-neutralizing antibodies (nAbs). NGS-based repertoire profiling will be used to obtain a quantitative readout of antibody response during immunization and to trace the lineage development of HCV nAbs. The antibody repertoire profiles will be used to guide immunogen optimization and to compare different immunogen candidates. The research proposed in Project 2 will therefore be complementary to and will benefit from the human sample analysis, epitope mapping, and testing of the Novartis E1E2 vaccine in Project 1, and together constitute the center for studying hepatitis C virus antibody responses and vaccine antigens.

项目概要据最新研究估计，全球约 3% 的人口感染了丙型肝炎病毒 (HCV)。已鉴定出 HCV E1 和 E2 糖蛋白上的许多中和表位，并用原子结构如何广泛地中和抗体 (bnAb) 与这些表位相互作用 2.65Å 晶体。 HCV E2 核心 (E2c) 与 bnAb (AR3C) 复合物的结构揭示了这种长期寻找的原子细节目标，为 HCV 疫苗的合理设计提供结构基础。在该 U19 提案的项目 2 中，我们。将结合 HCV 结构生物学的最新发现和计算领域的尖端技术 B 细胞库的蛋白质设计和下一代测序 (NGS)，以促进 HCV 免疫原设计在疫苗接种中诱导bnAbs的具体目标是：（1）开发针对表位的免疫原。和基于 E2 核心的免疫原与 bnAb 复合物中 E2 和 E1 中和表位的晶体结构。我们最近已经确定了呈现移植 HCV 的异源蛋白质支架。表位可以诱导针对该表位的交叉中和抗体。我们还发现 E2c 结构域可以。被优化为亚单位疫苗免疫原，以诱导针对保守的交叉中和抗体基于E2c的晶体结构，我们将缩短可变环，设计表面 N 连接聚糖，并向前层区域引入空间填充突变以稳定成功设计并表达的HCV抗原将在E2c构象上多价展示。铁蛋白颗粒和噬菌体 Qβ VLP (2) 使用 NGS 库分析来评估抗体反应。小鼠和非人类灵长类动物的免疫原将首先在小鼠中进行测试。与法律实验室合作，将提供并执行一系列 HCV 特异性免疫检测绘制一小组可诱导交叉中和的设计免疫原。西南国家灵长类研究中心将在非人类灵长类动物 (NHP) 中测试小鼠体内的抗体中心（SNPRC）将基于流式细胞术的单 B 细胞分选和基于微流体的 B 细胞封装。用于分离 HCV 中和抗体 (nAb)，并使用基于 NGS 的谱分析来获得。免疫期间抗体反应的定量读数并追踪 HCV 谱系发展 nAbs 抗体谱将用于指导免疫原优化并比较不同的抗体。因此，项目 2 中提出的研究将与候选免疫原互补并受益。来自项目 1 中诺华 E1E2 疫苗的人体样本分析、表位作图和测试，以及共同构成了研究丙型肝炎病毒抗体反应和疫苗抗原的中心。