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 开发针对快速进化的病原体（例如流感、艾滋病毒）的下一代疫苗需要实施 设计免疫原的合理方法可以将免疫反应引导至保守表位； 因此，应该会出现广泛、持久的反应，即“通用”或具有广泛保护性的流感疫苗。 在循环流感（例如 H1、H3）中诱导广泛的免疫力，并提供异亚型和大流行前的免疫力 项目 3 的目标是设计通过靶向引起这种保护性体液反应的免疫原。 我们在最初的基础上研究了流感血凝素（HA）和神经氨酸酶（NA）上的关键保守病毒位点。 免疫原设计策略主要集中于引导 B 细胞对 HA 受体结合位点的反应 (RBS) 来解决 T 细胞如何帮助影响引发的免疫反应。此外，我们整合了我们的免疫反应。 为 HA 建立免疫原设计策略并将其扩展到 NA，以了解设计是否有效 具体来说，我们使用我们的结构引导的原则来引导针对 HA 的免疫反应。 “表面重塑”方法将循环 N1 和 N2 NA 中的 NA 催化位点移植到外来的、禽类非 这些“受体”NA 将作为分子支架来呈现保守的 N1 和 N2。 催化位点但具有异源外围；引发的反应将保持保守的接触但适应 我们使用这些重新表面的 NA 来额外地扩展响应。 开发 NA 异嵌合四聚体 (NAtChs)，其呈现四个嫁接催化位点副本，但在四个 最后，我们将开发一种抗原性不同的支架，以增强对该表位的免疫聚焦。 使用定向进化平台的新型病毒-抗体协同进化测定来了解流感如何可能 进化以逃避我们的工程免疫原引起的集中免疫反应。 可能有助于预测病毒逃逸并允许识别疫苗方案，迫使流感进入 总的来说，该项目生成的数据将陷入“进化陷阱”。 有助于开发结合 HA 和 NA 的下一代流感疫苗 免疫原。