Adenoviral Vector-based Pandemic Influenza Vaccine

项目总结 H5、H7和H9亚型的低致病性和高致病性禽流感病毒都有能力 反复感染人类揭示了其人畜共患病的性质和大流行的可能性。此外，季节性流感 病毒(H1N1、H3N2和B型流感)继续演变，并在全球范围内构成重大公共卫生威胁。 虽然可以为个别流感毒株制造候选疫苗病毒，但准备疫苗是不切实际的。 每种可能的大流行病毒都有大量的疫苗储备。发展普适性的重要意义 流感疫苗对于制定更好的策略来对抗季节性和 潜在的大流行性流感病毒。 我们开发了一种新型复制缺陷牛腺病毒3型(BAd3)疫苗 平台，比目前可用的提供异源流感的Ad载体系统更好 通过剂量节约进行保护，不受预先存在的人类Ad载体免疫的影响。最近，我们 已发现不良疫苗平台可显著提高免疫原的表达水平 并与人Ad载体在小鼠体内的先天免疫和获得性免疫相关因子进行比较。我们还有 从猪瘟病毒CFP10蛋白中鉴定出一种22个氨基酸残基的自噬诱导多肽C5(AIP-C5 结核分枝杆菌可增强小鼠对H7N9流感病毒NP的强大T细胞免疫反应 通过广告载体交付。它提供了完全的保护(免于疾病、死亡或肺部病毒滴度)。 H1N1、H3N2H5N2H7N9和H9N2型流感病毒。重组腺病毒载体免疫小鼠的实验研究 H5N1 M2e-HA2[基质2的胞外区与血凝素(HA)的茎部分(HA2)连接，通过 HA信号肽和GCN4亮氨酸拉链三聚结构域]导致肺组织显著减少 H5N1病毒挑战后的病毒滴度。 这一建议是基于一种假设，即异亚型细胞介导免疫(CMI)的组合 对NP的反应和交叉反应(不一定是交叉中和)体液免疫反应 抗NP或其他保守结构域(M2e-HA2)，当用AIP-C5表达并通过 BAD疫苗平台，将为潜在的H5、H7或H9禽流感大流行提供广泛保护 病毒以及季节性的H1、H3和B型流感病毒。这项提议的目的是：i)调查 新型抗原设计和疫苗传递平台在小鼠模型中的免疫原性和保护作用 用于开发通用流感疫苗(目标1)；二)确定疫苗的免疫原性和保护效力 两种精选的雪貂通用流感疫苗配方(目标2)；和三)监测病毒传播 从接种疫苗的动物到未接种疫苗的动物，记忆B细胞和T细胞反应的质量， 保护性免疫和媒介免疫，以及可能与疫苗相关的加强呼吸系统疾病(目标3)， 利用最先进的技术。

PROJECT SUMMARY The ability of both low and highly pathogenic avian influenza (HPAI) viruses of H5, H7, and H9 subtypes to repeatedly infect humans reveals their zoonotic nature and pandemic potential. Besides, the seasonal influenza viruses (H1N1, H3N2 & influenza B) continue to evolve and pose significant public health threats worldwide. While candidate vaccine viruses can be made for individual influenza strains, it is impractical to prepare significant vaccine stocks for each of the potentially pandemic viruses. The significance of developing a universal influenza vaccine is of utmost importance for developing a better strategy for combatting seasonal as well as potential pandemic influenza viruses. We have developed a novel replication-defective bovine adenovirus (Ad) type 3 (BAd3)-based vaccine platform, which is better than the currently available Ad vector systems for providing heterologous influenza protection with dose sparing and is not impacted by the pre-existing human Ad vector immunity. Recently, we have revealed that the BAd vaccine platform induces significantly higher expression levels of the immunogen and innate and adaptive immunity-related factors compared to that of human Ad vectors in mice. We have also identified a 22 amino acid residues Autophagy-Inducing Peptide (AIP) C5 (AIP-C5) from the CFP10 protein of M. tuberculosis that enhances robust T cell immune responses in mice to NP of H7N9 influenza virus when delivered through an Ad vector. It conferred complete protection (from disease, death, or lung viral titers) against H1N1, H3N2, H5N2, H7N9, and H9N2 influenza viruses. Immunization of mice with an Ad vector expressing H5N1 M2e-HA2 [the extracellular domain of matrix 2 linked to the stem portion (HA2) of hemagglutinin (HA) with the HA signal peptide and the GCN4 leucine zipper trimerization domain] led to a significant reduction in lung viral titers following challenge with an H5N1 virus. This proposal is based on the hypothesis that a combination of heterosubtypic cell-mediated immune (CMI) responses against NP and the cross-reactive (not necessarily cross-neutralizing) humoral immune responses against NP or other conserved domains (M2e-HA2), when expressed with AIP-C5 and delivered through the BAd vaccine platform, will provide broad protection against potential pandemic H5, H7, or H9 avian influenza viruses as well as seasonal H1, H3, and influenza B viruses. The aims of this proposal are: i) To investigate immunogenicity and protective efficacy of novel antigen design and vaccine delivery platform in a mouse model for developing universal influenza vaccines (Aim 1); ii) To ascertain immunogenicity and protective efficacy of two selected universal influenza vaccine formulations in ferrets (Aim 2); and iii) To monitor virus transmission from the vaccinated to non-vaccinated animals, the quality of memory B and T cell responses, the durability of protective and vector immunity, and potential vaccine-associated enhancement of respiratory disease (Aim 3), utilizing the state-of-the-art technologies.