Start Date: May 01, 2013
End Date: May 01, 2014
Initially, we will test several different assays that evaluate cell-mediated immune responses, specifically interferon-gamma production in a recall assay by ELISA and ELISpot, using peripheral blood mononuclear cells or whole blood. We will also evaluate mucosal antibody in both nasal wash and oral fluids in a neutralization assay and whole-virus ELISA. We propose these types of assays because previous work indicates that intranasal SIV vaccines induce a strong cell-mediated immune response as well as antibody at the mucosa, as opposed to peripheral antibody in the sera. In addition, these sample types (peripheral blood, nasal wash and/or oral fluids) would be easy to collect and therefore producers would be more amenable to their collection. In addition, diagnostic labs already handle these specimen types. Initially pigs will be vaccinated by the intranasal route with either a live-attenuated IAV or replication-defective adenovirus encoding IAV HA and used for repeated sampling (blood, nasal wash and oral fluids). Once the assays have been established a subsequent study will be performed in which vaccinated pigs are challenged with heterologous virus to evaluate cross-protection and how protection correlates to responses detected in each assay.