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

项目摘要 这项跨学科的提案汇集了三项非常成功但独立的技术， 一种有效的、广泛中和的通用流感疫苗。我们的方法结合了免疫之息， 通过使用共识HA策略，与肯塔基州生物处理公司合作， (KBP)他们可以在植物中表达HA蛋白，并使用TMV-HA大大提高了亚单位疫苗的效力。 因此，本发明提供了一种缀合方法以产生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，我们希望我们的方法将推动长期免疫， 抗体和细胞免疫，为免疫保护的重叠机制提供了可能。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M2 e抗原。我们的具体目的如下：1）生产TMV-HA缀合物， 集中的H1、H3、H5基因和M2 e肽，2）免疫应答分析，鼠攻击研究和 3)TMV-HAc和TMV-M2 e疫苗的转化和雪貂研究。韦弗博士发明了一种 一种计算方法来表达HA的祖先序列，代表HA内序列的共有序列， 流感亚型。这些疫苗对漂移的季节性流感变种的保护效果优于 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 生产集中的HA蛋白的方法，具有为计划的研究生产蛋白的能力， 用于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)McCormick博士开发的一种改进HA亚单位疫苗的方法 力量TMV-HA疫苗还将与高度保守的M2 e肽疫苗组合，以拓宽 减少疫苗剂量。免疫学分析将用于确认疫苗效力， 以优化剂量、时间表和给药途径。疫苗效力和广泛的保护性免疫将 通过在疾病的鼠模型中使用9种不同病毒类型的致死性流感攻击来证实。最后， 疫苗配方将在雪貂中重新测试，雪貂是一种流感感染模型， 人类疾病的发展。我们的疫苗旨在驱动局部和全身免疫后， 鼻内或肌内给药途径，我们将使用免疫原性和病原体攻击 数据来定义优化的单剂量疫苗制剂。我们的目标是建立一个有效的普遍 可以大规模生产的流感疫苗，具有转化为 准备进行临床试验的通用疫苗产品。

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.