Universal Influenza DNA vaccine with Novel Toll-Like Receptor Adjuvants

描述（由申请人提供）：DNA疫苗接种通常提供的增强的安全性、稳定性和加速的产品开发使其成为开发“通用”流感疫苗的有吸引力的方法。不幸的是，灵长类动物的成功DNA疫苗接种需要用不期望的大剂量质粒进行多次接种。基因编码的佐剂可以提供必要的剂量节省效应，以实现实用的基于DNA的流感“通用”疫苗。我们打算利用显性阳性版本的Toll样受体（DP-TLR），以提供必要的免疫激活启动这样的保护性免疫反应的DNA疫苗接种。我们这些“概念验证”研究的疫苗靶标将是基质2（M2）和核蛋白（NP），这两种抗原在流感毒株之间表现出高度同源性，并已被广泛考虑用于此类“通用”疫苗。我们将通过以下具体目标来实现我们的目标：（1）构建TLR 2，TLR 4和TLR 9的DP版本，并评估它们激活NF-κ B的能力。B在人和小鼠体外基于细胞的测定中的作用;（2）在小鼠中评价与表达流感M2和NP的质粒共配制的DP-TLR的佐剂性和（3）在鼠病毒攻击研究中评价DP-TLR对表达流感M2和NP的质粒的剂量节省作用。与GM-CSF或IL-12相比，显著提高免疫原性或产生显著降低的保护剂量的佐剂构建体将在小鼠、雪貂和灵长类动物研究中作为II期SBIR应用中的高级流感DNA疫苗的组分进行进一步评价。公共卫生相关性：该项目的目标是开发新的佐剂，以支持和提高“通用”流感DNA疫苗的效力。这些佐剂将基于显性阳性的Toll样受体，通过NF-？B信令。

DESCRIPTION (provided by applicant): The enhanced safety, stability, and accelerated product development generally provided by DNA vaccination make it an appealing approach to develop a "universal" influenza vaccine. Unfortunately, successful DNA vaccination of primates requires multiple inoculations with undesirably large doses of plasmid. Genetically encoded adjuvants could provide the dose-sparing effect necessary to realize a practical DNA-based "universal" vaccine for influenza. We intend to exploit dominant-positive versions of Toll-like receptors (DP-TLRs) to provide the immune activation necessary to initiate such a protective immune response from DNA vaccination. Our vaccine targets for these "proof-of-concept" studies will be matrix 2 (M2) and nucleoprotein (NP), two antigens that exhibit a high degree of homology among strains of influenza and have been widely considered for such "universal" vaccines. We will pursue our objectives through the following specific aims: (1) construct DP-versions of TLR2, TLR4 and TLR9 and evaluate their capacity to activate NF-?B in human and mouse in vitro cell based assays; (2) evaluate the adjuvanticity of DP-TLRs co-formulated with plasmids expressing influenza M2 and NP in mice and (3) evaluate the dose-sparing effects of the DP- TLRs for the plasmids expressing influenza M2 and NP in murine viral challenge study. The adjuvant constructs that significantly improve immunogenicity or yield significant reductions in the protective dose over GM-CSF or IL-12 will be further evaluated in mice, ferret, and primate studies as components of an advanced influenza DNA vaccine in a Phase II SBIR application. PUBLIC HEALTH RELEVANCE: The objective of this project is to develop novel adjuvants that will support and improve the efficacy of a "universal" influenza DNA vaccine. These adjuvants will be based on dominant-positive toll-like receptors that activate through NF-?B signalling.