Nanoengineered virus-mimics as templates for design of a universal influenza A va

描述（由申请人提供）：流感病毒引起严重的呼吸道疾病，并有可能引起大流行。由于流感基因的高突变率，抗原漂移每年都会产生一种新的毒株。因此，每年监测病毒活动以及制造和向公众分发新的流感疫苗带来了巨大的经济负担。在甲型流感病毒粒子的表面发现了一个高度保守的膜蛋白（M2），它使自己成为开发通用流感疫苗的潜在靶标。然而，在自然情况下，M2的数量非常少（每个病毒颗粒16-22个），不能很好地暴露在病毒表面，免疫原性很差。我们克服这些挑战的方法是通过使用与不同功能肽结合的合成金（Au）纳米颗粒（AuNP）来模拟病毒结构，从而创建纳米工程病毒模拟物（NVM）。NVM可以高密度携带抗原。此外，其他功能肽也可以很容易地附着在NVM上。例如，可以附着能够使NVMs进入细胞或激活人体免疫细胞的肽。基于NVM的概念，我们建议开发一种通用的甲型流感疫苗递送系统。作为流感抗原，我们选择了流感a- a基质蛋白M2 （M2e）的高度保守的细胞外部分，作为细胞穿透肽，增强NVMs进入细胞，我们选择了来自人类免疫缺陷病毒（HIV-1）的（47-58）肽。我们的假设是，NVMs表面呈现高密度的M2e，并以其作为辅助肽，可以刺激广泛的保护性抗甲型流感免疫反应。本研究的目的是优化NVMs作为流感疫苗系统，并利用异源和异亚型流感毒株的活感染攻击来评估其在小鼠动物模型中的保护效果。重要的是，我们将评估鼻内免疫途径的全身和粘膜B细胞免疫。这个项目是新颖的，因为它试图利用纳米技术来创建一个模块化的疫苗输送系统，就像“乐高”积木一样，可以用来组装具有独特功能的nvm。这一概念可广泛用于创建针对多种传染病的疫苗输送系统。

DESCRIPTION (provided by applicant): Influenza virus causes serious respiratory illness and has potential to cause pandemics. Due to the high mutation rate in influenza genes, antigenic drift creates a new strain each year. Consequently there is significant economic burden to monitor virus activity and to create and distribute new influenza vaccines to the public each year. On the surface of the influenza A virion is found a highly conserved membrane protein (M2) which lends itself as a potential target for developing a universal influenza vaccine. However, under natural circumstances, M2 is present in very small numbers (16-22 per virus particles), is not well exposed at the virus surface and is poorly immunogenic. Our approach to overcome these challenges is to emulate virus structure through the use of a synthetic gold (Au) nanoparticle (AuNP) conjugated with different functional peptides creating a nanoengineered virus-mimic (NVM). The NVM can carry an antigen of choice in high density. In addition, other functional peptides can also be easily attached to the NVM. For example, peptides that can enable NVMs to enter cells or to activate the immune cells of the body could be attached. Based on the NVM concept, we propose to develop a universal influenza A vaccine delivery system. As the influenza antigen we have selected the highly conserved extracellular portion of the influenza-A matrix protein M2 (M2e), and as the cell-penetrating peptide to enhance entry of NVMs into cells we have selected tat (47-58) peptide derived from the human immunodeficiency virus (HIV-1). Our hypothesis is that NVMs presenting a high density of M2e on their surface with tat as a helper peptide can stimulate a broadly protective anti-influenza A immune response. The objectives of this study are to optimize NVMs as an influenza vaccine system and to evaluate their protective efficacy in mice animal models using live infectious challenges with heterologous and heterosubtypic influenza strains. Importantly we will evaluate systemic and mucosal B cell immunity for the intranasal routes of immunization. This project is novel because it seeks to exploit nanotechnology to create a modular vaccine delivery system, which like 'lego' pieces, can be used to assemble NVMs with unique functionalities. This concept can be used broadly used to create vaccine delivery systems against a host of infectious diseases. PUBLIC HEALTH RELEVANCE: This project focuses on development of a universal influenza A vaccine that can enable protection against all influenza A strains, thus eliminating the need for yearly vaccination against influenza. Successful completion of the project will reduce much morbidity, especially amongst elderly and children. It also has potential to be deployed on a mass scale as an anti-terror vaccine-agent against influenza.