2012 Annual Report
2. Characterize humoral and cellular immune responses to avian influenza viruses and identify epitopes associated with immunity.
3. Develop immunological reagents and methodologies to evaluate vaccine efficacy and protection.
The first objectives initiate the characterization of host cytokine expression profiles from specific innate immune cells in vitro. These profiles will then be compared with cytokine profiles obtained from avian influenza (AI) infected tissues, in vivo, for a better understanding of the overall innate immune response to avian influenza viruses (AIV). Innate immune cells will be isolated or produced for in vitro analysis from multiple sources, including: specific pathogen-free (SPF) White Leghorn (egg laying-type), SPF White Plymouth Rock (meat-type) chickens, and SPF Small Beltsville White SPF turkeys from the Southeast Poultry Research Laboratory. In addition, some experiments will utilize commercial chickens, turkeys, geese, ducks or others as needed, as sources for primary cell culture. We have in-house supplies of SPF birds and eggs, which can be used for these studies. In addition, we also have access to major histocompatibility complex (MHC)-defined birds (Avian Disease Oncology Laboratory, East Lansing, MI) which can be used for immunogenetic comparison of cytokine responses within members of Gallus gallus species. Expected outcomes for this objective will be the determination of which cytokines and transcription factors contribute to a resistant phenotype of avian species to AIV. It may be likely that different profiles are determined for different bird species. Once innate immunity profiles have been established, the cellular and humoral immune responses that contribute to natural and vaccine-induced protection will be characterized. We will utilize chickens, turkeys, ducks and other bird species in these studies. Experiments in Objective 2 will determine antibody levels following infection and vaccination, identify cellular immune responses to homologous and heterologous AI isolates, and determine putative epitopes involved with immunity. We will specifically address: (1) the induction of anti-viral antibodies that correlates with protective efficacy, decreases in virus shedding, and provide cross-reactivity to homo- and heterosubtypic isolates, (2) the induction of cellular immunity in poultry and wild birds following infection or vaccination and challenge, (3) identify T-cell epitopes to the hemagglutinin and nucleoprotein proteins of AI. Finally, we will develop immunological reagents and methodologies to evaluate vaccine efficacy and protection. Besides the usual indicators of vaccine induced protection, including survival, or decreases in shedding, assays to determine why and how a particular vaccine induces immunological protection against challenge are lacking. By incorporating cytokines and toll-like receptor agonists into vaccine formulations, their contributions to humoral and cellular immunity can be evaluated. Finally, the extent of cross protective immunity developed in vaccinated birds will be examined by utilizing antigenic cartography.
Kapczynski, D.R., Martin, A., Haddad, E.E., King, D.J. 2012. Protection from clinical disease against three highly virulent strains of Newcastle disease virus after in ovo application of an antibody-antigen complex vaccine in maternal antibody-positive chickens. Avian Diseases. 56(3):555-560. DOI: 10.1637/9980-110311-Reg.1.
Mesonero, A., Suarez, D.L., Van Santen, E., Tang, D., Toro, H. 2011. Avian influenza in ovo vaccination with replication defective recombinant adenovirus in chickens: Vaccine potency, antibody persistence, and maternal antibody transfer. Avian Diseases. 55:285-292.
Sylte, M.J., Suarez, D.L. 2012. Vaccination and acute phase mediator production in chickens challenged with low pathogenic avian influenza virus; novel markers for vaccine efficacy. Vaccine. 30(2012):3097–3105.
Hai, R., Garcia-Sastre, A., Swayne, D.E., Palese, P. 2011. A reassortment-incompetent live attenuated influenza virus vaccine for use in protection against pandemic virus strains. Journal of Virology. 85(14):6832-6843.
Cilloniz, C., Pantin Jackwood, M.J., Ni, C., Carter, V.S., Korth, M.J., Swayne, D.E., Tumpey, T.M., Katze, M.G. 2012. Molecular signatures associated with Mx-1 mediated resistance to highlyl pathogenic influenza virus infections: mechanisms of survival. Journal of Virology. 86:2437-2446.
Jiang, H., Yang, H., Kapczynski, D.R. 2011. Chicken interferon alpha pretreatment reduces virus replication of pandemic H1N1 and H5N9 avian influenza viruses in different avian species lung cell cultures. Virology Journal.8:447.