Project 3: Influenza Virus HA and NA Immunogen Design

项目3：流感病毒HA和NA免疫原设计

• 批准号: 10549613
• 负责人: Aaron Gregory Schmidt
• 金额: $ 36.16万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10549613
• 关键词: Address; Affect; Affinity; Antibodies; Antigen-Antibody Complex; Antigens; B cell repertoire; B-Lymphocyte Epitopes; B-Lymphocytes; Binding Sites; Biochemical; Biological Assay; Biophysics; Birds; Catalytic Domain; Chimera organism; Coupling; Cysteine; Data; Development; Directed Molecular Evolution; Engineering; Epitopes; Escape Mutant; Evolution; Exposure to; Generations; Genes; Goals; HIV; Hemagglutinin; Homo; Human; Immune; Immune response; Immunity; Immunization; In Vitro; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza Hemagglutinin; Island; Libraries; Macaca mulatta; Mammalian Cell; Massachusetts; Modification; Molecular; Monoclonal Antibodies; Mus; Mutagens; Mutation; Neuraminidase; Outcome; Pattern; Property; Reaction; Recombinant Proteins; Recombinants; Regimen; Role; Site; Structure; Structure of germinal center of lymph node; Surface; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Vaccines; Variant; Viral; Viral Antibodies; Virus; Yeasts; arms race; design; exhaustion; human subject; imprint; in vivo; influenza virus vaccine; innovation; movie; neutralizing antibody; next generation; novel; novel vaccines; pathogen; pre-pandemic; programs; receptor binding; response; scaffold; viral RNA; viral fitness; viral resistance

Abstract Developing next-generation vaccines for rapidly evolving pathogens (e.g., influenza, HIV) requires implementing rational approaches to engineer immunogens that can direct immune responses to conserved epitopes; in doing so, broad, durable responses are likely to occur. A “universal” or broadly protective influenza vaccine should induce broad immunity within circulating influenzas (e.g., H1, H3) and provide heterosubtypic and pre-pandemic protection. A goal of Project 3 is to design immunogens that elicit such protective humoral responses by targeting key conserved viral sites on the influenza hemagglutinin (HA) and neuraminidase (NA). We build upon our initial immunogen design strategies primarily focused on directing B-cell responses to the HA receptor binding site (RBS) to address how T-cell help influences the elicited immune responses. Furthermore, we integrate our immunogen design strategies established for HA and extend them to NA in order to understand if the design principles for directing immune responses to HA are generalizable. Specifically, we use our structure-guided “resurfacing” approach to graft the NA catalytic site from circulating N1 and N2 NAs onto exotic, avian non- circulating NAs. These “acceptor” NAs will serve as molecular scaffolds to present the conserved N1 and N2 catalytic site but with a heterologous periphery; elicited responses will maintain conserved contacts but adapt to the heterologous periphery and thus broadening the response. We use these resurfaced NAs to additionally develop NA heterochimeric tetramers (NAtChs) that present four copies of the grafted catalytic site but on four antigenically distinct scaffolds in efforts to increase immune focusing to this epitope. Finally, we will develop a novel viral-antibody coevolution assay using directed evolution platforms to understand how influenza may evolve to escape a focused immune response elicited by our engineered immunogens. This innovative approach may help anticipate viral escape and allow identification of vaccine regimen(s) that force influenza into an “evolutionary trap” by compromising overall viral fitness. Collectively, the data generated within this Project will contribute to developing next-generation influenza vaccines incorporating both HA and NA as potential immunogens.

摘要 为快速演变的病原体(如流感、艾滋病毒)开发下一代疫苗需要实施 合理的方法设计免疫原，引导免疫反应到保守的表位；在做 因此，可能会出现广泛、持久的反应。一种“通用”或具有广泛保护性的流感疫苗应该 在循环中的流感病毒(如H1、H3)中诱导广泛的免疫，并提供异型和大流行前的免疫 保护。项目3的一个目标是设计免疫原，通过靶向诱导这种保护性体液反应 流感血凝素(HA)和神经氨酸酶(NA)上的关键保守病毒位点。我们在最初的基础上 免疫基因设计策略主要集中于将B细胞反应定向到HA受体结合部位 (苏格兰皇家银行)，以解决T细胞如何帮助影响引发的免疫反应。此外，我们将我们的 为HA建立了免疫基因设计策略，并将其扩展到NA，以了解设计是否 将免疫反应定向到HA的原则是可以概括的。具体地说，我们使用我们的结构指导 将循环中的N1和N2NA的NA催化部位移植到外来的，非禽类的 循环中的NAS。这些“受体”将作为分子支架呈现保守的n1和n2。 催化部位，但具有异源外围；激发的反应将保持保守的接触，但适应 异源外围，从而扩大了反应范围。我们使用这些重新浮出水面的NAS来另外 开发NA异构体四聚体(NAtChs)，其呈现四个副本的接枝催化中心，但在四个 抗原性不同的支架，努力增加对该表位的免疫聚焦。最后，我们将开发一种 使用定向进化平台的新型病毒-抗体协同进化分析以了解流感如何 进化以逃避由我们的工程免疫原引发的集中免疫反应。这一创新方法 可能有助于预测病毒逃逸，并允许识别迫使流感进入 “进化陷阱”，因为它损害了病毒的整体适应性。总而言之，本项目内生成的数据将 帮助开发下一代流感疫苗，将HA和NA并列为潜在疫苗 免疫原。