2012 Annual Report 1a.Objectives (from AD-416): Identify functional genetic variations that modulate the immune responses to swine mucosal pathogens; and determine the genetic profiles of ”good responders” to swine vaccines. 1b.Approach (from AD-416): Out-bred commercial breeds of swine will be vaccinated with recombinant products from bacterial and viral pathogens by various routes including mucosal exposure with novel and conventional adjuvants and vaccine platforms. Local and systemic responses to vaccination will be evaluated by measuring antigen specific antibody isotype responses in the serum, and eye, lung, and intestinal fluids, and local and whole blood changes in cytokine and chemokine biomarkers by gene expression. This information will be used by both ARS and the COOPERATOR to jointly develop novel adaptive and acquired immune readout systems to evaluate vaccine efficacy as it applies to swine directly and as a second species for testing important new vaccine constructs. 3.Progress Report: Pigs were immunized at different sites with soluble protein antigens with and without adjuvant and serum antibody isotypes specific for the test protein, and then were evaluated by an enzyme-linked immunosorbent assay. Although immunization via the conjunctiva, by mouth, in the peritoneum, and intradermal routes produced measurable serum antibodies of all isotypic classes, skin site sensitization was most immunogenic for the induction of specific antibodies (IgE). This response can be used to evaluate allergic disease responses to immunization because IgE is the antibody that links responses to protein to immediate hypersensitivity diseases.