Expression of the avian-specific toll-like receptor 15 in chicken heterophils is mediated by Gram-negative and Gram-positive bacteria, but not TLR agonists

Toll-like receptors (TLRs) are a critical component of the innate immune response of mammalian and avian species. While most mammalian TLRs have been well characterized, the chicken-specific TLR15 has not been extensively studied. We recently demonstrated that TLR15 is differentially expressed between Salmonella-susceptible-and-resistant chickens, indicating a potential role in the innate immune response to infection with Salmonella. The aim of the present study was to gain better insight into the nature of the ligand for TLR15 by characterizing gene expression patterns of TLR15 by heterophils in response to numerous bacterial-derived TLR agonists LPS, flagellin, CpG oligodeoxynucleotides, lipotechoic acid (LTA), peptidoglycan (PGN), and Pam3CSK4 (PAM), stimulation with live Salmonella enterica serovar Enteritidis (SE-used as a positive control), chicken isolates of Escherichia coli (EC) and Enterococcus gallinarum (EG), the equine-specific pathogen Rhodococcus equi, and stimulation with heat-killed, and formalin-killed SE, EC, and EG. TLR15 expression increased significantly in response to stimulation with live, heat-killed and formalin-killed SE, EC, and EG, but was unaffected by stimulation with known TLR agonists and R. equi. Overall, these observations demonstrate that the individual TLR agonists are not the ligand for TLR15, and that TLR15 recognizes a unique, non-secreted, heat-stabile component of both Gram-negative and Gram-positive bacteria commonly found in and/or capable of causing disease in chickens.