Structure based design of trimer interface epitope focused universal influenza vaccines

基于三聚体界面表位的通用流感疫苗的结构设计

基本信息

批准号：
10361516
负责人：
James E Crowe
金额：
$ 122.11万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-12 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10361516
关键词：
Affinity Antibodies Antibody Repertoire Antigens B-Lymphocytes Binding Binding Sites Biological Biological Products Blood Cells Cells Complex Computer Models Crystallization Crystallography Development Electron Microscopy Engineering Epitopes Exposure to Generations Genes Glycoproteins Goals Hand Head Hemagglutinin Human Immune Immune response Immunity In Vitro Individual Infection Influenza Influenza Hemagglutinin Influenza vaccination Knowledge Laboratories Linear Programming Machine Learning Maps Masks Mass Spectrum Analysis Memory B-Lymphocyte Methods Modeling Nature Polysaccharides Population Proteins Research Research Project Grants Roentgen Rays Sequence Analysis Site Standardization Structure Techniques Testing Therapeutic antibodies Vaccine Antigen Vaccine Design Vaccines Validation Viral Viral Vaccines Virus Virus Diseases X-Ray Crystallography antibody engineering base combat cross reactivity design experimental study immunogenicity in silico in vivo influenza virus strain influenza virus vaccine influenzavirus innovation laboratory experiment molecular recognition mouse model multidisciplinary nanoparticle neutralizing antibody next generation next generation sequencing novel pandemic disease particle programs protective efficacy rational design receptor binding response scaffold screening structural biology therapeutic vaccine tool universal influenza vaccine vaccine candidate

项目摘要

The “Computational Models of Immunity” projects in this application focus on development and implementation of new structure-based design tools for influenza hemagglutinin (HA) protein trimer interface specific antibodies or vaccine antigens. These projects will use knowledge about the structure and function of human neutralizing antibodies to the trimer interface of the HA head that we have in hand or will discover, in order to design new antibodies or vaccines in silico. We have access to peripheral blood cells from a diverse panels of subjects with prior natural infection, or exposure to experimental inoculation with vaccines encoding HA molecules with both seasonal vaccines and unusual experimental influenza subtypes, including H3variant, H5, H6, H7, H9, and H10 viruses. The immune B memory cell populations from these individuals are the ideal starting materials to isolate unusual heterosubtypic antibodies. Recently, we identified the HA head trimer interface as a major new site of vulnerability for universal influenza antibodies and candidate vaccines. Here, we will study existing and isolate additional broadly heterosubtypic human antibodies to the trimer interface of the HA head. We will determine the immunome of the responding heterosubtypic clones using high-throughput next generation sequencing of antibody gene repertoires that comprise the clonal lineages of the most heterosubtypic antibodies isolated. Once antibodies with unusual breadth or activity are isolated, the structure of these antibodies will be determined in complex with purified HA molecules in the Structural Core using crystallography and single particle electron microscopy (EM) studies. Such structures will provide the coordinates for the modeling experiments using Rosetta. We will in silico mature human antibodies to increase affinity for the HA antigen of specific virus types and use multi-state design to maximize breadth, i.e., create antibodies that recognize HAs of all clades, subtypes, groups, or even types. We then will synthesize and express these novel antibodies and determine neutralization activity, binding affinity, and competition binding groups of designed antibodies, using a diverse HA panel and pseudotyped viruses with all type A HAs in nature. The co-crystal structure of these human antibodies with HA will be the template for in silico design of structurally stable epitope-focused immunogens. We will first validate these designed immunogens by testing the interaction with the target human antibodies. Further, these immunogens will be experimentally tested by evaluating immune responses. Then, we will use the novel immunogens to isolate new antibodies from subjects naturally exposed to influenza, to show that the immunogens present antigens recognized by natural immune responses.

本申请中的“免疫计算模型”项目重点关注开发和实施新的基于结构的流感血凝素（HA）蛋白设计工具这些项目将使用有关三聚体界面特异性抗体或疫苗抗原的知识。 HA 头三聚体界面的人中和抗体的结构和功能我们已经掌握或将要发现的信息，以便在计算机上设计新的抗体或疫苗。可以获得来自先前自然感染的不同受试者的外周血细胞，或暴露于编码HA分子的疫苗的实验接种，具有季节性疫苗和不寻常的实验性流感亚型，包括 H3 变种、H5、H6、H7、H9 和来自这些个体的 H10 病毒免疫 B 记忆细胞群是理想的起点。最近，我们鉴定出了 HA 头三聚体。界面作为通用流感抗体和候选者的主要新脆弱点在这里，我们将研究现有的并分离更多的广泛异亚型人类。 HA 头三聚体界面的抗体我们将确定 HA 头的免疫组。使用高通量下一代抗体测序响应异亚型克隆包含分离的大多数异亚型抗体的克隆谱系的基因库。一旦分离出具有异常广度或活性的抗体，这些抗体的结构将与结构核心中纯化的 HA 分子复合使用来确定晶体学和单粒子电子显微镜（EM）研究将提供此类结构。使用 Rosetta 进行建模实验的坐标我们将在计算机上模拟成熟的人类。抗体以增加对特定病毒类型的 HA 抗原的亲和力并使用多状态设计以最大化广度，即创建识别所有进化枝的 HA 的抗体，然后我们将合成并表达这些新的抗体。并确定设计的中和活性、结合亲和力和竞争结合基团抗体，使用不同的 HA 组和具有自然界中所有 A 型 HA 的假型病毒。这些人类抗体与 HA 的共晶结构将成为计算机设计的模板我们将首先验证这些设计的结构稳定的表位免疫原。此外，通过测试与目标人类抗体的相互作用来识别免疫原。然后，我们将通过评估免疫反应来对免疫原进行实验测试。新的免疫原可以从自然暴露于流感的受试者中分离出新的抗体，显示免疫原呈现自然免疫反应识别的抗原。