Rapid Manufacturing of a Universal Flu Vaccine Using TMV-conjugated Centralized Antigens

项目摘要 这个跨学科的提案汇集了三种非常成功但独立的技术来创造 一种有效且广泛中和的通用流感疫苗。我们的方法结合了免疫力的呼吸， 通过使用共识 HA 策略、生产速度和产能 与 Kentucky BioProcessing 合作 (KBP)，可以在植物中表达 HA 蛋白，并使用 TMV-HA 大大提高了亚单位疫苗的效力 结合方法创建候选通用流感病毒疫苗。根据我们的临床前数据 三种甲型流感毒株，我们预计我们的方法将推动长期免疫，在 抗体和细胞免疫，提供了免疫保护重叠机制的潜力。我们 汇集不同学科的强大团队，共同目标是快速展示概念验证 H1、H3、H5 和 M2e 抗原。我们的具体目标如下： 1) 生产 TMV-HA 缀合物 集中的 H1、H3、H5 基因和 M2e 肽，2) 免疫反应分析、小鼠攻击研究和 3) TMV-HAc 和 TMV-M2e 疫苗的转化和雪貂研究。韦弗博士开发了一种 表达 HA 祖先序列的计算方法，代表内部序列的共识 流感亚型。这些疫苗比流感疫苗更好地预防季节性流感变异 传统的三价灭活病毒疫苗。与 KBP 合作，我们将使用已建立的植物表达 生产集中HA蛋白的方法，具有生产用于计划研究的蛋白质的能力， 以供未来的临床试验开发。 HA共有蛋白将融合到烟草花叶病毒的表面 (TMV) 通过化学缀合，麦考密克博士开发的一种改进 HA 亚单位疫苗的方法 效力。 TMV-HA 疫苗还将与高度保守的 M2e 肽疫苗结合使用，以拓宽 保护并减少疫苗剂量。将使用免疫学分析来确认疫苗的效力，以便 优化剂量、时间表和给药途径。疫苗功效和广泛的保护性免疫力将 通过在小鼠疾病模型中使用 9 种不同病毒类型的致命流感攻击来证实。最后， 疫苗配方将在雪貂身上进行重新测试，雪貂是一种更接近于模拟流感病毒感染的模型 人类疾病的进展。我们的疫苗旨在在以下任一情况后驱动局部和全身免疫： 鼻内或肌内给药途径，我们将利用免疫原性和病原体攻击 数据来定义优化的单剂量疫苗配方。我们的目标是建立一个有效的通用 可以大规模生产的流感疫苗，具有转化为疫苗的巨大潜力 准备进行临床测试的通用疫苗产品。

Project Abstract This interdisciplinary proposal brings together three highly successful but independent technologies to create an effective and broadly neutralizing universal influenza vaccine. Our approach combines breath of immunity, by using a consensus HA strategy, production speed and capacity by working with Kentucky BioProcessing (KBP), who can express HA protein in plants, and vastly improved subunit vaccine potency using a TMV-HA conjugation approach to create a candidate universal influenza virus vaccine. Based on our preclinical data on three influenza A strains, we expect our approach will drive long term immunity that is balanced between antibody and cellular immunity, providing potential for overlapping mechanisms of immune protection. We bring together a strong team of different disciplines with a combined goal to quickly show proof of concept with H1, H3, H5, and M2e antigens. Our Specific Aims are as follows: 1) Production of TMV-HA conjugates of centralized H1, H3, H5 genes and M2e peptide, 2) Immune response analysis, murine challenge studies and 3) Translational and ferret studies of TMV-HAc and TMV-M2e vaccines. Dr. Weaver has developed a computational method to express an ancestral sequence of HA, representing a consensus of sequences within an Influenza subtype. These vaccines protect against drifted seasonal influenza variants better than a traditional trivalent inactivated virus vaccine. In partnership with KBP, we will use established plant expression methods to produce centralized HA proteins, with the capacity to produce protein for the planned studies and for future clinical trial development. HA consensus protein will be fused to the surface of Tobacco Mosaic virus (TMV) by chemical conjugation, a method developed by Dr. McCormick to improve HA subunit vaccine potency. TMV-HA vaccines will also be combined with a highly conserved M2e peptide vaccine, to broaden protection and reduce vaccine dose. Immunological analysis will be used to confirm vaccine potency, in order to optimize dose, schedule and route of administration. Vaccine efficacy and broadly protective immunity will be confirmed by lethal influenza challenge using 9 divergent virus types in a murine model of disease. Finally, vaccine formulations will be re-tested in ferrets, a models of influenza infection which more closely mimics the progress of disease in humans. Our vaccine is designed to drive local and systemic immunity after either intranasal or intramuscular routes of administration, and we will use immunogenicity and pathogen challenge data to define an optimized single dose vaccine formulation. Our goal is to generate an effective universal vaccine against influenza that can be manufactured at scale, with significant potential for translation into a universal vaccine product that is ready for clinical testing.