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

描述（由申请方提供）：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生产和向公众分发新流感疫苗是巨大的经济负担。在甲型流感病毒粒子的表面上发现了高度保守的膜蛋白（M2），其本身适合作为开发通用流感疫苗的潜在靶标。然而，在自然条件下，M2以非常小的数量存在（每个病毒颗粒16-22个），不能很好地暴露在病毒表面，免疫原性差。我们克服这些挑战的方法是通过使用与不同功能肽缀合的合成金（Au）纳米颗粒（AuNP）来模拟病毒结构，从而产生纳米工程病毒模拟物（NVM）。NVM可以以高密度携带选择的抗原。此外，其他功能性肽也可以容易地附着到NVM。例如，可以附着能够使NVM进入细胞或激活身体免疫细胞的肽。基于NVM的概念，我们建议开发一种通用的甲型流感疫苗输送系统。作为流感抗原，我们选择了流感-A基质蛋白M2（M2 e）的高度保守的细胞外部分，并且作为增强NVMs进入细胞的细胞穿透肽，我们选择了源自人免疫缺陷病毒（HIV-1）的达特（47-58）肽。我们的假设是，在其表面上呈现高密度的M2 e的NVMs与作为辅助肽的达特可以刺激广泛的保护性抗甲型流感免疫应答。本研究的目的是优化NVMs作为流感疫苗系统，并评估其在小鼠动物模型中的保护效力，使用异源和异亚型流感病毒株的活感染性攻击。重要的是，我们将评价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”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许多传染病的疫苗递送系统。 公共卫生相关性：该项目的重点是开发一种通用的甲型流感疫苗，可以保护所有甲型流感病毒株，从而消除对 每年接种流感疫苗。该项目的成功完成将大大降低发病率，特别是老年人和儿童的发病率。它也有可能被大规模部署作为一种针对流感的反恐疫苗剂。

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.