Start Date: Jan 03, 2006
End Date: Jan 02, 2011
1) Assess innate immune variability in chickens and turkeys by examining the frequency of genetic polymorphism in genes related to innate immunity (cytokines/chemokines, toll-like receptors [TLR]). Initial analysis will be undertaken in 2 pedigree lines of chickens that we have characterized in terms of innate immune function and disease susceptibility and commercial lines of turkeys and their wild turkey counterparts; 2) Identify differentially expressed or genetically altered genes in heterophils from the innate immune functionally divergent lines of chickens and turkeys by use of suppressive subtractive hybridization (SSH). Normalized, directionally cloned avian heterophil cDNA libraries will be constructed from pools of mRNA purified from resting, inflammatory, and activated heterophils following either the in vitro stimulation with inflammatory agonists or in vivo following infection with Salmonella or Campylobacter; 3) Develop new modulators of innate immunity such as pathogen associated molecular patterns (PAMPs) as immune modulators of early colonization, PAMPs as adjuvants for commercial live vaccines, and the development of new live vaccines that specifically induce a heterophil-mediated innate immune response; 4) Apply anti-sense and RNAi technologies in poultry by introducing anti-sense oligonucleotides and RNAi to available cell lines (the macrophage HD-11 cell line) by transfection, using western-blot and Real Time-PCR to assess the effectiveness of gene silencing, and characterizing the function of the target gene by stimulating the cells with PAMPs and measuring immune responses. The in vivo experimental approach is similar to that stated above, except that in ovo route will be used to introduce anti-sense oligonucleotides and RNAi; and 5) Evaluate anti-microbial peptides from chicken and turkey heterophils that have potential as new biotherapeutics for food-borne bacteria.