Universal Influenza DNA vaccine with Novel Toll-Like Receptor Adjuvants

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 vaccinati on 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 do minant-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 nucl eoprotein (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 TL R2, 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 ef fects 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 fur ther 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 ef ficacy of a universal influenza DNA vaccine. These adjuvants will be based on dominant-positive toll-like receptors that activate through NF-?B signalling.