SBIR-STTR Award

A universal influenza vaccine is believed to be possible if conserved regions of influenza are effectively targeted and appropriate immune responses are generated against those targets. The enhanced safety, stability, and accelerated product development generally provided by DNA vaccination make it an appealing approach to develop such a universal influenza vaccine. Unfortunately, immune responses to universal influenza antigens are typically weak and in the past, DNA vaccination of humans has been disappointing. To overcome these and other obstacles to developing an effective, practical, and truly universal influenza vaccine, we intend to deliver our vaccine using a DNA prime / protein boost regimen and employ novel immunogens derived from the following three conserved influenza A antigens: 1) the stem region of hemagglutinin (HA); 2) the matrix 2 protein ectodomain (M2e); and, 3) the nucleoprotein (NP). We believe that together, these antigens will evoke the immunological breadth necessary to protect against a broad range of both seasonal and potential pandemic influenza strains. We will also use a potent DNA adjuvant combination to maximize immunogenicity and to tune the responses toward a Th1 phenotype. Finally, we will utilize a recombinant protein boost to amplify the humoral immune responses and increase their durability. In this Phase I SBIR, we will construct and express our influenza A immunogens and verify their immunogenicity and protective efficacy in mice to determine if the vaccine provides a wide breadth of protection from divergent seasonal and pandemic influenza A strains. If we are successful in this Phase I proof-of-concept study, we will move on to test our vaccine in a macaque challenge model under a Phase II application and begin development on influenza B, and possibly type C, immunogens.

Public Health Relevance Statement:
PROJECT NARRATIVE The objective of this project is to develop a DNA prime / subunit boost universal influenza vaccine. Such a vaccine is needed to combat seasonal and pandemic influenza outbreaks and to lessen the dependency on seasonal vaccines.

NIH Spending Category:
Biodefense; Biotechnology; Emerging Infectious Diseases; Genetics; Immunization; Infectious Diseases; Influenza; Pneumonia & Influenza; Prevention; Vaccine Related

Project Terms:
Address; Adjuvant; Algorithms; Animals; Antibodies; Antibody Response; antibody-dependent cell cytotoxicity; Antigens; base; Birds; Body Weight decreased; Cessation of life; Codon Nucleotides; combat; Combined Vaccines; Consensus; cytokine; Data; Databases; Dependency; design; Development; Disease; DNA; DNA Sequence; DNA Vaccines; Elderly; Epidemic; Equilibrium; expression vector; Family suidae; Future; Health; Hemagglutinin; Hospitals; Human; Human Resources; Immune response; Immunity; Immunization; immunogenicity; in vivo; Infection; Inflammatory; Influenza; Influenza A virus; Influenza B Virus; influenza outbreak; influenza virus vaccine; influenzavirus; Life; Macaca; Measures; Modeling; Morbidity - disease rate; Mus; neutralizing antibody; novel; Nucleoproteins; Outcome Study; pandemic influenza; Pathogenesis; Phase; Phenotype; Phylogenetic Analysis; Population; product development; protective efficacy; Protein Subunits; Proteins; Recombinant Proteins; Recombinants; Regimen; response; Risk; Safety; seasonal influenza; Secondary Immunization; Small Business Innovation Research Grant; Source; stem; Subunit Vaccines; T cell response; Testing; Vaccinated; Vaccination; Vaccine Antigen; vaccine candidate; Vaccines; Variant; Viral; Viral Load result; Wages; Western Blotting

A universal influenza vaccine is believed to be possible if conserved regions of influenza are effectively targeted and appropriate immune responses are generated against those targets. The enhanced safety, stability, and accelerated product development and production generally provided by DNA vaccination make it an appealing approach to develop such a universal influenza vaccine. Unfortunately, immune responses to universal influenza antigens are typically weak and earlier studies of DNA vaccination in humans showed poor humoral immunogenicity. However, recent advancements in vaccine delivery and use of adjuvants and viral vector boosts have improved the outlook of DNA vaccination. To overcome obstacles to developing an effective, practical, and truly universal influenza vaccine, we propose to further develop and test our DNA prime / viral-vectored boost vaccine regimen, which employs novel immunogens derived from the following four conserved influenza A antigens: 1) the stem region of hemagglutinin (Stem); 2) the matrix 2 protein ectodomain (M2e); 3) the nucleoprotein (NP); and 4) the matrix 1 protein (M1). Our phase I results indicate that together, these antigens evoke the immunological breadth and types of immune responses necessary to target a broad range of both seasonal and potential pandemic influenza strains and to evoke appropriate antiviral immune responses necessary to combat these influenza viral infections. Our strategy also uses a clinically proven DNA adjuvant that maximizes immunogenicity, tunes the responses toward an antiviral Th1 phenotype, and improves the anti- viral activities of the humoral immune response. Under this phase II SBIR application, we will test a viral-vectored boost for amplification of the DNA-raised immune responses. We will also determine the protective effects of our vaccine in other relevant preclinical animal models (ferrets and macaques) and perform studies intended to further define the vaccine's mechanisms of action. 3

Public Health Relevance Statement:
Project Narrative The objective of this project is to continue development of a “universal” influenza A vaccine. Such a vaccine is needed provide protection against all influenza A viruses to a substantial portion of the world's population.

Project Terms:
Adjuvant; Animal Model; Animals; Antibodies; Antibody Formation; Antigens; Antiviral Agents; booster vaccine; CD8-Positive T-Lymphocytes; Clinical; combat; Development; DNA; DNA amplification; DNA Vaccines; Fc Receptor; Ferrets; Hemagglutinin; Human; Immune response; Immunize; immunogenic; immunogenicity; Immunologics; improved; Individual; Inflammatory; Influenza; Influenza A virus; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Interferon Type II; Interleukin-12; Lung; Macaca; Mediating; Mucous Membrane; Mus; nonhuman primate; novel; Nucleoproteins; pandemic influenza; Pathogenicity; Phase; Phenotype; Population; pre-clinical; Pre-Clinical Model; product development; Production; protective effect; protective efficacy; Proteins; Publishing; Reagent; Recombinants; Regimen; response; Role; Safety; Small Business Innovation Research Grant; stem; T cell response; T-Lymphocyte Subsets; Testing; Time; Toxic effect; universal influenza vaccine; Vaccinated; Vaccination; vaccine delivery; Vaccines; vector; vector vaccine; Vesicular stomatitis Indiana virus; Viral Pathogenesis; Viral Physiology; Viral Vector; Virus; Virus Diseases