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：
10576343
负责人：
James E Crowe
金额：
$ 121.34万
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/10576343
关键词：
Affinity Antibodies Antibody Repertoire Antigen Presentation Antigens B-Lymphocytes Binding Binding Sites Biological Biological Products Blood Cells Cells Complex Computer Models Crystallography Development Electron Microscopy Engineering Epitope Mapping Epitopes Experimental Models 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 Mass Spectrum Analysis Memory B-Lymphocyte Methods Modeling Nature Polysaccharides Population Proteins Reaction Research Research Project Grants Roentgen Rays Seasons 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 combat cross reactivity design design verification immunogenicity in silico in vivo influenza virus strain influenza virus vaccine influenzavirus innovation laboratory experiment molecular recognition monomer mouse model multidisciplinary nanoparticle neutralizing antibody next generation next generation sequencing novel pandemic potential 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.

本应用程序中的“免疫计算模型”项目的重点是开发和用于影响型hemagglutin（HA）蛋白的新的基于结构的设计工具三聚体界面特异性抗体或疫苗抗原。这些项目将使用有关的知识人类中和抗体的结构和功能为了在计算机中设计新的抗体或疫苗，我们已经有或会发现。我们有能力从具有先前自然感染的受试者的潜水员面板中获取外周血细胞，或与编码HA分子的疫苗接触实验接种，这两个季节疫苗和异常的实验影响亚型，包括H3Variant，H5，H6，H7，H9和 H10病毒。这些人的免疫B记忆细胞种群是理想的开始分离出异常的杂型抗体的材料。最近，我们确定了Ha Head Trimer 界面是通用影响力抗体和候选者的主要脆弱性新地点疫苗。在这里，我们将研究现有的并隔离额外的广泛异质型人类 HA头的触发界面的抗体。我们将确定使用高通量的下一代抗体响应异源性克隆完成分离的最异性替代抗体的克隆谱系的基因曲线。一旦孤立了具有异常宽度或活性的抗体，这些抗体的结构将使用在结构核中使用纯化的HA分子在复合物中确定晶体学和单颗粒电子显微镜（EM）研究。这样的结构将提供使用Rosetta进行建模实验的协调。我们将在计算机成熟的人类中增加对特定病毒类型的HA抗原的亲和力的抗体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抗体，亚型，组甚至类型。然后，我们将合成并表达这些新型抗体并确定设计的神经元化活性，结合亲和力和竞争结合组使用潜水员HA面板和A型A类具有ASTY AS的抗体。这这些人类抗体与HA的共结晶结构将是用于计算机设计的模板结构稳定的以表位为中心的免疫原。我们将首先验证这些设计通过测试与靶标人类抗体的相互作用来进行免疫原子。此外，这些免疫原子将通过评估免疫回报对实验测试。然后，我们将使用新型免疫原以分离新抗体与自然暴露于影响力的受试者，向表明免疫原的抗原具有天然免疫回报识别的抗原。