Temporal dynamics of innate and adaptive immune responses in broiler birds to oral delivered chitosan nanoparticle-based Salmonella subunit antigens

Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) infection of poultry causes a significant risk to public health through contamination of meat and eggs. Current Salmonella vaccines have failed to provide strong mucosal immunity in the intestines to reduce Salmonella shedding and food contamination. Considering the short lifespan of broilers, an easy-to-deliver, safe and effective Salmonella vaccine is urgently needed. Our goal in this study was to demonstrate the ability of chitosan nanoparticle (CNP) vaccine delivery platform in activating immune response to Salmonella antigens in broilers inoculated orally. In an initial study, soluble whole antigen of SE entrapped in CNP was inoculated but the specific immune responses were poor. Therefore, the CNP entrapped immunogenic outer membrane proteins (OMP) and flagellin (FLA) of SE and surface conjugated with FLA [CNP-(OMP + FLA)] was developed. In broilers inoculated orally with CNP-(OMP + FLA) formulation once or twice, we monitored the temporal expression of innate immune molecules and antigen specific lymphocyte proliferation. In the cecal tonsils of CNP-(OMP + FLA) inoculated birds, we observed enhanced expression of mRNA coding Toll-like receptors (TLRs)- 1, 4, 5, and 7, especially at dpv 21. In addition, both OMP and FLA specific lymphocytes proliferation at dpv 7 and 21 by CNP-(OMP + FLA) were enhanced in the spleen. In conclusion, CNP-(OMP + FLA) formulation augmented both innate and lymphocyte responses in orally inoculated broilers. Further studies are needed to determine the candidate subunit CNP vaccine’s efficacy in a challenge trial.