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.

抽象的 开发用于快速发展的病原体（例如，影响力，艾滋病毒）的下一代疫苗需要实施 可以将免疫反应构成表位的工程剂免疫原理的理性方法；做 因此，可能会发生广泛的耐用响应。 “通用”或广泛保护的影响应疫苗 在循环影响力（例如H1，H3）中诱导广泛的免疫力 保护。项目3的目标是设计免疫原，通过靶向来引起这种受保护的体液反应 在影响扎血凝集素（HA）和神经瘤酶（NA）上的关键配置病毒位点。我们以最初的 免疫原设计策略主要集中于指导B细胞对HA受体结合位点的反应 （RB）解决T细胞如何有助于影响引起的免疫反应。此外，我们整合了 为HA建立的免疫原设计策略并将其扩展到NA，以了解该设计是否 指导对HA的免疫反应的原则是可推广的。具体而言，我们使用我们的结构引导 从循环N1和N2 NAS移植到异国情调的非鸟类非 - 循环NAS。这些“受体” NAS将用作分子支架，以呈现配置的N1和N2 催化部位，但具有异源外周；引起的响应将保持保守的联系，但适应 异源外周，从而扩大了反应。我们使用这些重新浮出水面的NAS 开发Na杂聚聚物四聚体（Natchs），这些四聚体呈现移植催化位点的四个副本，但在四个 抗原上不同的支架，以增加对该表位的免疫聚焦。最后，我们将发展一个 新型病毒 - 抗体协同进化分析使用定向的进化平台，以了解影响力可能如何 进化是为了逃避我们工程的免疫原子引起的重点免疫反应。这种创新的方法 可能有助于预测病毒逃逸，并允许识别疫苗疗法的疫苗，从而影响疫苗 总体病毒适应性受损“进化陷阱”。总的来说，该项目中生成的数据将 有助于发展下一代影响编码HA和NA作为潜力的疫苗 免疫原子。