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

项目摘要 这项跨学科的计划将三项非常成功但独立的技术结合在一起，以创建 一种有效和广泛中和的通用流感疫苗。我们的方法结合了免疫的气息， 通过使用一致的HA战略，通过与肯塔基生物加工公司合作，实现生产速度和产能 (KBP)，谁可以在植物中表达HA蛋白，并使用TMV-HA极大地提高了亚单位疫苗的效力 接合方法创造候选通用流感病毒疫苗。根据我们的临床前数据 三种甲型流感病毒株，我们预计我们的方法将推动长期免疫，在 抗体和细胞免疫，为重叠的免疫保护机制提供了可能性。我们 汇聚一个由不同学科组成的强大团队，具有共同的目标，以快速展示概念验证 H1、H3、H5和M2e抗原。我们的具体目标如下：1)生产TMV-HA偶联物 集中的H1、H3、H5基因和M2e肽，2)免疫反应分析，小鼠挑战研究和 3)TMV-HAc和TMV-M2e疫苗的翻译和雪貂研究。韦弗博士已经开发出一种 表示HA的祖先序列的计算方法，表示内序列的一致性 一种流感亚型。这些疫苗对漂移的季节性流感变种的预防效果比 传统三价灭活病毒疫苗。与KBP合作，我们将使用已建立的植物表达 方法生产集中的HA蛋白，具有为计划的研究和 用于未来的临床试验开发。HA共识蛋白将与烟草花叶病毒表面融合 化学偶联(TMV)是McCormick博士开发的一种改进HA亚单位疫苗的方法 威力。TMV-HA疫苗还将与高度保守的M2e多肽疫苗结合，以扩大 保护和减少疫苗剂量。将使用免疫学分析来确认疫苗的效力，以便 优化给药剂量、给药程序和给药途径。疫苗的效力和广泛的保护性免疫将 在小鼠疾病模型中，使用9种不同的病毒类型进行致命流感攻击，证实了这一点。最后， 疫苗配方将在雪貂身上重新测试，雪貂是一种流感感染模型，更接近于 人类疾病的进展。我们的疫苗旨在促进局部和系统免疫，无论是 鼻腔或肌肉内给药途径，我们将使用免疫原性和病原体挑战 定义优化的单剂疫苗配方的数据。我们的目标是产生一个有效的普遍 可规模化生产的流感疫苗，具有转化为 已准备好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.