Phase I SBIR proposals will be accepted. Fast-Track proposals will not be accepted. Phase I clinical trials will not be accepted. Number of anticipated awards: 1 Budget (total costs): Phase I: up to $243,500 for up to 6 months; Phase II of up to $1,000,000 and a Phase II duration of up to 2 years PROPOSALS THAT EXCEED THE BUDGET OR PROJECT DURATION LISTED ABOVE MAY NOT BE FUNDED. Background Influenza is the cause of considerable morbidity and mortality globally resulting in an estimated 290,000-650,000 fatalities annually and is a pathogen of significant public health importance. Vaccination remains the most effective measure against influenza infection. In addition to annual epidemics, pandemics are also a major concern as demonstrated by three major influenza pandemics in the 20th century in 1918, 1957 and 1968, and the first influenza pandemic of the 21st century in 2009 that spread worldwide in a short period, causing significant morbidity and mortality. In addition, circulation and infection of humans with novel avian influenza viruses from subtypes H5N1, H7N7, H7N1, H7N3, H7N9, and H9N2 has occurred. Vaccination remains the most effective measure against influenza infection. However, currently available vaccines include egg- or cell-derived inactivated split or live attenuated vaccines or insect cell-derived recombinant hemagglutinin (HA) protein. Apart from antibody responses to the globular head region of HA, antibody responses to the HA stalk, neuraminidase (NA), M2e and cell-mediated immune (CMI) responses to conserved internal proteins such as nucleoprotein (NP) have been shown to play a major role in viral clearance. Currently available vaccines are standardized based on HA content, although inactivated detergent-split vaccines do contain variable amounts of other viral proteins and the antibody responses to them varies. Furthermore, inactivated vaccines are not efficient in inducing/recalling CMI responses. Hence, a vaccine that induces antibody responses to all known antibody targets of influenza virus and, when delivered as nanoparticles, induces CMI responses to major conserved internal proteins, is needed to provide both the depth and breadth required. Project Goals The primary objective is to develop a recombinant protein and/or peptide-based influenza vaccine that can induce both humoral and cell-mediated immune responses, with sufficient breadth and depth to major antibody and CMI target proteins, when delivered with appropriate nanoparticles. The nanoparticle technology and recombinant protein/peptide synthesis process should be scalable with batch-to-batch consistency. Phase I Activities and Expected Deliverables 1. Develop a recombinant protein and/or peptide-based vaccine that contains HA, NA, M1, stalks of HA and the conserved protein, NP, from H1N1, H3N2 and B (Yamagata and Victoria lineages) viruses. 2. Demonstrate induction of antibody responses to HA, NA, M2e, stalks of HA and CMI responses to NP induced by the candidate vaccine as compared to those induced by a licensed, inactivated vaccine in appropriate animal models, mice and/or ferrets. 3. Compare immune responses induced by candidate vaccine by intranasal vs intramuscular routes in animal models, mice and/or ferrets. Page 132 Impact Currently available influenza vaccines induce strain-specific antibody responses against the vaccine strains included in the vaccine. Furthermore, they are poor inducers of CMI responses. If there is a mismatch between the circulating strain/s and vaccine strains, the vaccine efficacy will be suboptimum. Hence, a vaccine that induces a broader antibody and CMI responses to confer protection against disease, reduces morbidity, viral loads and symptoms will have a major impact on public health. Commercialization Potential Currently, there are no licensed recombinant protein/peptide influenza vaccines that contain, apart from HA, defined amounts of NA, M1 and NP to induce both humoral and cell-mediated immune responses. Hence, a vaccine that induces antibody responses to HA, NA, M2e, and stalk, and CMI responses to conserved internal protein NP, would increase the needed breadth and depth of vaccination and would have tremendous commercialization potential as this will provide broader protection from disease, even when the circulating strains of viruses are different from those contained in the vaccine.