Novel broad spectrum vaccination against influenza

描述（由申请人提供）：Cure Lab发现了一种疫苗设计的新方法，可提供针对流感感染的广谱免疫力，流感经常影响世界人口，并造成相当大的医疗保健和经济负担。每年，世界人口的5-15%感染流感，导致300 - 500万例严重疾病，估计有250，000 - 500，000人死于流感相关并发症。这笔赠款的目的是开发新一代流感疫苗并推向市场。在这个阶段，我们将在动物模型中证明我们的方法的原理，并开发产品原型。我们将修改流感病毒保守的内部NP和M1蛋白，使它们在免疫后产生增强的T细胞应答。这将通过改变所述蛋白质的构象来实现，使得泛素-蛋白酶体系统更有效地降解它们，从而产生更多的MHC I类呈递肽，诱导更有效和长期的T细胞应答。首先，我们将构建表达修饰的A型流感NP基因的质粒和重组牛痘病毒，并在小鼠模型中测试保护效力和免疫原性。其次，我们将测试流感病毒A和B的其他保守内部蛋白的蛋白酶体依赖性降解，根据Cure Lab算法修饰鉴定的靶蛋白，并分析其细胞内降解，从而制备它们用作我们疫苗方案的一部分。第三，构建表达去稳定化保守流感蛋白的多种疫苗载体，并在体内测试其免疫原性和保护效力，建立沿着的免疫保护相关方式，以及最佳免疫途径和时间表。该项目的总体目标是证明与野生型病毒蛋白相比，使用修饰的流感蛋白作为免疫原具有更好的保护作用或/和增强的免疫应答。

DESCRIPTION (provided by applicant): Cure Lab has discovered a novel approach to vaccine design that provides broad spectrum immunity against influenza infection, influenza regularly affects the world population and imposes a considerable health care and economic burden. Annually, 5-15% of the world's population contracts influenza, resulting in 3-5 million cases of severe illness, and an estimated 250,000 to 500,000 deaths from influenza-associated complications. The purpose of this grant is to develop and bring to market a new generation influenza vaccine. In this phase I grant, we will establish proof of principle for our approach in an animal model and develop a product prototype. We will modify conserved internal NP and M1 proteins of influenza virus in such a way that they will generate enhanced T-cell response upon immunization. This will be accomplished by changing the conformation of said proteins in such a way that the ubiquitin-proteasome system degrades them more efficiently and thus produces more MHC class l-presented peptides, inducing a more effective and long-term T-cell response. First, we will construct both a plasmid and a recombinant vaccinia virus expressing a modified NP gene of influenza A and test protective efficacy and immunogenicity in the mouse model. Second, we will test proteasome-dependent degradation of additional conserved internal proteins of influenza viruses A and B, modify identified target proteins according to Cure Lab algorithm and analyze their intracellular degradation, thus preparing them for use as a part of our vaccine regimen. Third, we will construct multiple vaccine vectors expressing destabilized conserved influenza proteins and test their immunogenicity and protective efficacy in vivo, establishing along the way the immune correlates of protection, as well as the best immunization routes and schedules. The overall aim of this project is to demonstrate better protective effect or/and enhanced immune response using modified influenza proteins as immunogens compared to wild-type viral proteins.