Co-Presentation and Delivery of TLR Agonist Combinations with Subunit Antigens to Pathogen-Match the Immune Response

项目总结/摘要 亚单位疫苗是预防传染病和相关死亡的关键策略。但使用 与完整病原体相比，亚单位抗原引起较弱的免疫应答， 安全和有效的佐剂，以提高疫苗效力，增加免疫应答，并减少大小和 免疫力所需的疫苗数量。然而，我们缺乏一个详细的图片， 先天性免疫受体同时或依次在同一细胞或不同细胞上， 免疫系统的反应。我们的研究团队设计了一种新的，灵活的，模仿病原体的 能够一起呈递功能性抗原的重组外膜囊泡（rOMV）平台 与所需的佐剂组合。我们打算利用这项创新技术来研究如何 TLR和NOD激动剂的不同佐剂组合参与先天免疫系统，并帮助引导免疫应答。 对病毒和细菌病原体的免疫应答。我们的假设，将通过 特异性目的是rOMV可以与佐剂的特异性组合共呈递抗原以诱导免疫应答。 病原体匹配和保护性免疫应答（例如，抗病毒和抗细菌）。具体目标 目的1：鉴定由单个佐剂或佐剂组合对rOMV引起的分子途径。 将用TLR和NOD激动剂修饰rOMV，以了解所产生的先天性免疫受体是如何被激活的。 激动剂使用已建立的报告细胞激活和影响细胞信号传导途径，并确定这些 通过TLR和NOD激动剂的共呈递改变了信号通路。我们将比较可溶性 在一些实施方案中，相同佐剂的组合对信号传导和树突状细胞活化和成熟的影响是显著的。目标二： 确定rOMV-佐剂组合如何指导使用OVA免疫的适应性免疫应答 作为模型抗原。我们将使用OVA抗原来表征rOMV诱导的适应性免疫应答 表达单一佐剂或佐剂的组合，并将它们与单一可溶性佐剂或可溶性佐剂的组合进行比较。 我们将鉴定促进强的和定向的T细胞、B细胞和/或记忆应答的组合。 目的3：确定定制的rOMV佐剂组合是否可以指导特异性抗病毒和抗细菌 保护性免疫我们将使用OVA呈递的rOMV，以及在细胞表面呈递的病原体特异性抗原。 在病毒（流感）和病毒（流感）的激发模型中，用不同的单一或组合佐剂修饰的rOMV 和经修饰以表达OVA的细菌病原体（李斯特菌）以及天然病原体（即，无OVA）。我们 将研究先天信号通路激活和免疫反应如何与保护性免疫相关。 所提出的研究的预期结果是对佐剂组合如何 在分子、细胞和生物体水平上发挥作用，以防止病毒和细菌病原体。成果 这将有助于确定一套指导原则，我们和该领域的其他调查人员可以更有力地利用这些原则， 设计佐剂系统以最大化亚单位疫苗的功效。

Project Summary/Abstract Subunit vaccines are a key strategy for preventing infectious disease and related deaths.. However, the use of subunit antigens, which elicit a weaker immune response than intact pathogen, has coincided with a need for safe and effective adjuvants to improve vaccine efficacy, increase immune response, and reduce the size and quantity of vaccine necessary to impart immunity. However, we lack a detailed picture of how engaging multiple innate immune receptors simultaneously or sequentially, on the same cell, or different cells, alters the outcome of the immune response. Our research team has engineered a new, flexible, pathogen-mimicking recombinant outer membrane vesicle (rOMV) platform capable of presenting functional antigen together with desired combinations of adjuvants. We intend to use this innovative technology to investigate how different adjuvant combinations of TLR and NOD agonists engage the innate immune system and help direct the immune response against viral and bacterial pathogens. Our hypothesis, to be evaluated through the Specific Aims, is that rOMVs can co-present antigen with specific combinations of adjuvants to induce pathogen-matched and protective immune responses (e.g., anti-viral and anti-bacterial). The specific aims are to: Aim 1: Identify the molecular pathways elicited by individual or combinations of adjuvants on rOMVs. rOMVs will be decorated with TLR and NOD agonists to understand how the resulting innate immune receptor agonists activate and influence cell signaling pathways using established reporter cells, and determine how those pathways are altered by co-presentation of TLR and NOD agonists. We will compare the response to soluble combinations of the same adjuvants on both signaling and dendritic cell activation and maturation. Aim 2: Determine how rOMV-adjuvant combinations direct the adaptive immune response to immunization using OVA as a model antigen. We will use OVA antigen to characterize the adaptive immune response induced by rOMV expressing single or combinations of adjuvants and compare them to single or combinations of soluble adjuvants. We will identify the combination(s) that promote strong and directive T cell, B cell and/or memory responses. Aim 3: Determine whether tailored rOMV adjuvant combinations can direct specific anti-viral and anti-bacterial protective immunity. We will use OVA-presenting rOMVs, as well as pathogen-specific antigens presented on rOMVs, decorated with different single or combined adjuvants in challenge models for both a viral (influenza) and a bacterial pathogen (Listeria) modified to express OVA as well as the native pathogens (i.e., no OVA). We will examine how innate signaling pathway activation and immune response correlate to protective immunity. The anticipated outcome of the proposed studies is a mechanistic understanding of how adjuvant combinations work at the molecular, cellular and organismal level to protect against viral and bacterial pathogens. The outcome will help to define a set of guiding principles that we and other investigators in the field can use to more robustly design adjuvant systems to maximize the efficacy of subunit vaccines.