Novel broad spectrum vaccination against influenza

描述(由申请人提供)：Cure Lab发现了一种新的疫苗设计方法，可以提供对流感感染的广谱免疫，流感经常影响世界人口，并施加相当大的医疗保健和经济负担。每年，世界上有5%-15%的人口感染流感，导致300万-500万例严重疾病，估计有25万-50万人死于与流感相关的并发症。这笔赠款的目的是开发新一代流感疫苗并将其推向市场。在这一阶段，我们将在动物模型中为我们的方法建立原则证明，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细胞反应。这将通过改变这些蛋白质的构象来实现，使泛素-蛋白酶体系统更有效地降解它们，从而产生更多MHC类L呈现的多肽，诱导更有效和更长期的T细胞反应。首先，我们将构建表达甲型流感病毒修饰NP基因的重组痘苗病毒和重组痘苗病毒，并在小鼠模型中测试保护效果和免疫原性。其次，我们将测试流感病毒A和B的额外保守内部蛋白的蛋白酶体依赖的降解，根据Cure Lab算法修改识别的目标蛋白，并分析它们的细胞内降解，从而为用作我们疫苗方案的一部分做好准备。第三，我们将构建多个表达不稳定保守流感蛋白的疫苗载体，并在体内测试它们的免疫原性和保护效果，建立沿途保护的免疫相关因素，以及最佳免疫路线和免疫程序。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效果或/和增强免疫反应。

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.