Development of a Novel Universal Influenza Vaccine

To determine the effectiveness of R-DOTAP as an adjuvant for recombinant COBRA H1 and H3 antigens, provided by Dr. Ted Ross, in the elicitation of a broad antibody response against historic and current circulating strains of IAV. Preliminary analytical development and formulation studies will be undertaken to establish effective conditions for mixture of the R-DOTAP nanoparticles and the protein antigens and enable characterization and monitoring of the stability of candidate vaccine mixtures. This will allow production of vaccine candidates to study the appropriate R-DOTAP/antigen ratios and dosing to generate maximal antibody and T-cell responses in animal studies. Mice will be vaccinated with two different H3 and one H1 COBRA antigens adjuvanted with R-DOTAP or squalene based adjuvants and sera tested in HAI assays with a variety of historic and current influenza strains Hypothesis: Based on these novel properties, we predict that H3 and H1 recombinant COBRA antigens adjuvanted with R-DOTAP will be superior to other adjuvants for the elicitation of antibodies that neutralize a broad number of co-circulating viral variants in both H1 and H3 subtypes. Specific Aim 2: To determine the ability of R-DOTAP to promote CD8 T cell responses to recombinant influenza nucleoprotein. Mice will be vaccinated with recombinant nucleoprotein adjuvanted with R-DOTAP or squalene based adjuvants and CD8 T cell responses will be evaluated by ELISPOT assay. Hypothesis: R-DOTAP will promote strong CD8 T cell responses to conserved epitopes within the nucleoprotein sequence. Specific Aim 3: To determine if a mixture of COBRA HA and nucleoprotein adjuvanted with R-DOTAP will generate antibody responses to HA and CD8 T cell responses to nucleoprotein conserved epitopes in a single vaccine. Mice will be vaccinated with a mixture of COBRA HA antigens and recombinant nucleoprotein. Sera and cells will be assayed by HAI and ELISPOT assays respectively. Hypothesis: Vaccination with COBRA HA + NP adjuvanted with R-DOTAP will promote both antibody and CD8 T cell responses.