Universal Influenza DNA vaccine with Novel Toll-Like Receptor Adjuvants

描述（由申请人提供）：DNA疫苗通常提供增强的安全性、稳定性和加速的产品开发，使其成为开发“通用”流感疫苗的一种有吸引力的方法。不幸的是，成功的灵长类动物DNA疫苗接种需要多次接种不受欢迎的大剂量质粒。基因编码佐剂可以提供必要的剂量节约效应，以实现实用的基于dna的流感“通用”疫苗。我们打算利用toll样受体（DP-TLRs）的显性阳性版本来提供必要的免疫激活，以启动DNA疫苗接种的这种保护性免疫反应。我们这些“概念验证”研究的疫苗靶点将是基质2 （M2）和核蛋白（NP），这两种抗原在流感毒株中表现出高度同源性，已被广泛认为是这种“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TLR2、TLR4和TLR9的dp版本，并评估它们激活NF-？B在人和小鼠体外细胞基础上的测定；(2)评估DP-TLRs与表达流感M2和NP的质粒共配制的小鼠佐剂性；(3)在小鼠病毒攻毒研究中评估DP-TLRs对表达流感M2和NP的质粒的剂量节约作用。与GM-CSF或IL-12相比，显著提高免疫原性或显著降低保护剂量的佐剂结构将在小鼠、雪貂和灵长类动物研究中进一步评估，作为II期SBIR应用的高级流感DNA疫苗的组成部分。公共卫生相关性：该项目的目标是开发新型佐剂，以支持和提高“通用”流感DNA疫苗的功效。这些佐剂将基于通过NF-?激活的显性阳性toll样受体B信号。

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.