Novel Influenza nano vaccines for broad cross protection

总结 流感是一个重大的公共卫生风险。目前的季节性流感疫苗对密切 在健康成年人中匹配病毒，但它不能防止流行病或大流行病的爆发， 流感病毒经常变异，人畜共患病毒株可跨越物种障碍进入人类。其他 季节性流感疫苗的缺点包括每个季节都需要生产新的疫苗， 选择疫苗株的不确定性，以及对不匹配病毒的功效受损。一种新型 称为通用流感疫苗的流感疫苗的产生将克服这些挑战。在 在上一个资助期，我们已经从保守的HA茎中制备了层状蛋白质纳米颗粒（纳米簇 抗原和甲型流感的M2蛋白胞外域。纳米簇免疫诱导的交叉保护 针对甲型流感两个系统发育组的病毒，包括潜在的大流行禽流感毒株。 甲型流感和B型流感都可引起人类流感流行。在本提案中，我们建议 构建由新设计的抗原蛋白组成的多价层状纳米簇制剂， 流感A和流感B作为通用流感疫苗。新疫苗将引起广泛的交叉感染- 预防两种流感。我们有三个具体目标： 目标1。设计和构建甲型和B流感保守抗原， 从这些和以前设计的抗原蛋白质，并表征这些新的纳米簇。我们 将优化这些纳米团簇的编排、组成和稳定性，以用于生理激活的 游离抗原蛋白的释放，树突细胞摄取后的抗原加工和呈递， 这些纳米簇分布到引流淋巴结，并诱导强抗原特异性免疫 小鼠的反应。 目标2.为了测试这些层状纳米团簇或最佳组合是否会广泛诱导 反应性免疫反应以及免疫力是否会提供针对病毒的交叉保护 在小鼠中跨越甲型流感和B型流感。 目标3：为了测试领先的多价纳米簇组合是否会诱导强大的免疫反应， 反应，赋予广泛的交叉保护雪貂。 总的来说，我们的研究将开发出一种广泛交叉保护的通用流感疫苗。

SUMMARY Influenza is a major public health risk. The current seasonal influenza vaccine is effective against closely matched viruses in healthy adults, but it cannot prevent the outbreaks of epidemics or pandemics because influenza viruses mutate frequently and zoonotic strains can jump the species barrier into humans. Other disadvantages of the seasonal influenza vaccine include the need to produce new vaccines every season, the uncertainty in selecting vaccine strains, and the compromised efficacy for mismatched viruses. A novel generation of influenza vaccines, termed universal influenza vaccines, will overcome these challenges. In the previous grant period, we have produced layered protein nanoparticles (nanoclusters) from conserved HA stalk antigens and the M2 protein ectodomain of influenza A. Nanocluster immunizations induced cross protection against viruses from both phylogenic groups of influenza A, including pandemic-potential avian strains. Both influenza A and influenza B cause influenza epidemics in humans. In this proposal, we propose to construct a multivalent layered nanocluster formulation composed of newly designed antigenic proteins from both influenza A and influenza B as a universal influenza vaccine. The new vaccine will induce broad cross- protection against both influenza types. We have three specific aims: Aim 1. To design and construct conserved antigens from influenza A and B, fabricate nanoclusters from these and previously designed antigenic proteins, and characterize these new nanoclusters. We will optimize the orchestration, composition, and stability of these nanoclusters for the physiologically-activated release of free antigenic proteins, antigen-processing and presentation after the uptake by dendritic cells, distribution of these nanoclusters to draining lymph nodes, and induction of strong antigen-specific immune responses in mice. Aim 2. To test whether these layered nanoclusters or an optimal combination will induce broadly reactive immune responses and whether the immunity will grant cross-protection against viruses spanning both influenza A and influenza B in mice. Aim 3. To test whether the leading multivalent nanocluster combinations will induce robust immune responses which confer broad cross-protection in ferrets. Overall, our research will develop a broadly cross-protective universal influenza vaccine.