CpG oligonucleotides and recombinant interferon-γ in combination improve protection in chickens to Salmonella enterica serovar Enteritidis challenge as an adjuvant component, but have no effect in reducing Salmonella carriage in infected chickens.

Salmonella enterica serovar Enteritidis is the most common cause of human salmonellosis in many developed nations. It is frequently associated with both poultry meat and eggs. In the present study we have determined whether CpG oligonucleotides that stimulate the immune system via Toll like-receptors 15 and 21 in the chicken can be used as immunomodulatory agents to break carriage of S. Enteritidis in in vitro and in vivo infection models. We also investigated its use as a component in an adjuvant to stimulate cell mediated immunity with a killed vaccine preparation. Following infection of the chicken macrophage-like cell line HD11 with Salmonella enterica serovar Gallinarum, cells were stimulated with an oligonucleotide containing a CpG motif, or with a non-CpG oligonucleotide control at concentrations ranging from 0 to 80 µM. Addition of the CpG oligonucleotide greatly enhanced clearance of S. Enteritidis in dose-dependent manner, whilst the control oligonucleotide had no significant effect. In contrast, stimulation of cells infected with S. Gallinarum had no effect. The CpG or control oligonucleotide with recombinant chicken interferon-γ was administered intramuscularly into chickens experimentally colonized with S. Enteritidis, although neither preparation produced any change in intestinal colonization levels to that in untreated control birds. Finally, CpG oligonucleotides were incorporated with recombinant interferon-γ, double-stranded RNA (Poly I:C) and squalene as a Th1-stimulating combined adjuvant for synergistic activation of cellular immunity (CASAC) together with whole killed Salmonella as the antigen as an experimental vaccine. Following vaccination and challenge of chickens with S. Enteritidis, CASAC gave significant protection to intestinal colonization whereas the same antigen given with a proprietary adjuvant did not. Neither adjuvant increased protection to systemic infection. The data suggest that adjuvants incorporating CpG motifs and interferon-γ may improve protection afforded by killed-Salmonella vaccines.