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

• 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
• 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

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”项目侧重于开发和 流感病毒血凝素（HA）蛋白基于结构设计新工具的实现 三聚体界面特异性抗体或疫苗抗原。这些项目将利用有关知识， HA头部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，以便通过计算机设计新的抗体或疫苗。我们 可以接触到来自不同组的先前有自然感染的受试者的外周血细胞， 或暴露于编码HA分子的疫苗的实验性接种， 疫苗和不常见的实验性流感亚型，包括H3变异型、H5、H6、H7、H9和 H10病毒。来自这些个体的免疫B记忆细胞群是理想的开始 分离不寻常的异亚型抗体的材料。最近，我们鉴定了HA头部三聚体， 作为通用流感抗体和候选物主要新的脆弱性位点的界面 疫苗。在这里，我们将研究现有的和分离的其他广泛的异亚型人类 抗体与HA头部的三聚体界面接触。我们将确定 使用高通量下一代抗体测序的应答异亚型克隆 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宽度或活性的抗体，这些抗体的结构 将在结构核心中与纯化的HA分子复合， 晶体学和单粒子电子显微镜（EM）研究。这些结构将提供 使用罗塞塔进行建模实验的坐标。我们将在电脑上成熟的人类 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 设计以最大化宽度，即，产生识别所有进化枝的HA的抗体， 子类型、组甚至类型。然后我们将合成并表达这些新的抗体 并确定设计的抗体的中和活性、结合亲和力和竞争结合基团， 抗体，使用不同的HA组和具有所有A型HA的假型病毒。的 这些人抗体与HA的共晶体结构将是用于在计算机上设计HA的模板。 结构稳定的表位聚焦免疫原。我们将首先验证这些设计 通过测试与靶向人抗体的相互作用来确定免疫原。此外，这些 免疫原将通过评估免疫应答进行实验测试。然后，我们将使用 从自然暴露于流感的受试者中分离新抗体的新免疫原， 表明免疫原呈递天然免疫应答识别的抗原。