Universal T cell targeted influenza vaccine

摘要 流感每年在全世界造成显著的发病率和死亡率，并引起严重的 当新的病毒株进化出以前没有在人类中传播时，就会发生大流行。病毒的高变异率 需要根据每年流行的毒株的流行情况来生产新的疫苗。这些 流感疫苗的改良版本并不总是具有保护作用;疫苗有效性（VE） 在过去10年中，根据疫苗株与流行株的匹配程度，从10%到60%。 对于诱导更大的交叉保护性免疫的流感疫苗存在迫切的未满足的需求。我们 建议利用广泛反应性流感特异性T细胞表位的免疫原性潜力， 具有普遍意义的疫苗。我们之前的R21资助工作提供了原理证明， 先进的免疫信息学工具可用于有效鉴定高度保守的甲型流感表位， 包括混杂的CD 4 + T细胞表位和HLA-A2限制的CD 8 + T细胞表位， 免疫原性，并可诱导保护性免疫。此外，我们令人信服地证明， 设计用于刺激人T细胞应答的基于T细胞的疫苗可以诱导异型保护性T细胞免疫应答。 免疫力我们现在建议扩展我们的R21研究，以更全面地评估混杂的CD 4 + T细胞 表位，以证实这些表位可以在>95%的所有人类中引发有效的CD 4 + T细胞应答 表达不同的HLA II类等位基因。我们还将鉴定相关的CD 8 + T细胞表位， 额外的非HLA-A2 I类超型，以获得足够的表位用于广泛的人群覆盖（>95%的 人类）。然后，我们将开发和比较多表位疫苗的免疫原性和保护效力 使用几种最先进的疫苗输送平台，包括：重组“裸”DNA，纯化蛋白质， 与新型佐剂混合，设计用于逃避预先存在的人Ad的新型腺病毒（Ad）疫苗 免疫和病毒样颗粒（VLP）包被RNA疫苗。疫苗将在体外进行测试， 人PBMC和体内使用表达转基因HLA的人源化小鼠。异型功效将是 在用3种不同的甲型流感病毒株（H1N1、H3 N2和H5 N1）攻击后进行评价。拟议工作 可为流感疫苗开发提供转型性新产品和方向， 一个关于诱导广泛保护性T细胞反应的范式转变概念。

Abstract Influenza is responsible for significant morbidity and mortality worldwide every year and causes severe pandemics when new strains evolve that have not previously circulated in humans. The high viral mutation rate necessitates that new vaccines be generated based on the prevalence of circulating strains every year. These reformulated versions of influenza vaccines are not always protective; vaccine effectiveness (VE) has varied from 10 to 60% over the past 10 years based on how well vaccine strains are matched with circulating strains. There is an urgent unmet need for influenza vaccines that induce greater cross-protective immunity. We propose to harness the immunogenic potential of broadly reactive influenza-specific T cell epitopes to produce vaccines with universal significance. Our previous R21 funded work has provided proof-of-principle that advanced immunoinformatic tools can be used to efficiently identify highly conserved influenza A epitopes, including promiscuous CD4+ T cell epitopes and HLA-A2-restricted CD8+ T cell epitopes, that are immunogenic and can induce protective immunity. Furthermore, we have convincingly demonstrated that T cell-based vaccines designed to stimulate human T cell responses can induce heterotypic protective immunity. We now propose to extend our R21 studies to more fully evaluate promiscuous CD4+ T cell epitopes to confirm that these epitopes can elicit potent CD4+ T cell responses in >95% of all humans expressing diverse HLA class II alleles. We also will identify relevant CD8+ T cell epitopes restricted by additional non-HLA-A2 class I supertypes, to obtain sufficient epitopes for broad population coverage (>95% of humans). We then will develop and compare immunogenicity and protective efficacy of multi-epitope vaccines using several state-of-the art vaccine delivery platforms including: recombinant ‘naked’ DNA, purified proteins mixed with novel adjuvants, novel adenovirus (Ad) vaccines designed to evade preexisting human Ad immunity, and virus-like particle (VLP) encapsidated RNA vaccines. Vaccines will be tested in vitro using human PBMC and in vivo using humanized mice expressing transgenic HLA. Heterotypic efficacy will be evaluated upon challenge with 3 distinct influenza A strains (H1N1, H3N2, and H5N1). The proposed work can provide transformational new products and direction for influenza vaccine development, focusing on a paradigm-shifting concept of inducing broadly protective T cell responses.