title: Extracellular adenosine 5'-triphosphate and lipopolysaccharide induce interleukin-1β release in canine blood.
abstract: Binding of extracellular adenosine 5'-triphosphate (ATP) or lipopolysaccharide (LPS) to the damage-associated molecular pattern receptor P2X7 or the pathogen-associated molecular pattern receptor Toll-like receptor (TLR)4, respectively, can induce the release of the pleiotropic cytokine interleukin (IL)-1β in humans and mice. However, the release of IL-1β in dogs remains poorly defined. Using a canine IL-1β enzyme-linked immunosorbent assay, this study investigated whether ATP or LPS could induce IL-1β release in a canine blood-based assay. Short-term incubations (30 min) with ATP induced IL-1β release in LPS-primed canine blood, and this process could be near-completely impaired by the P2X7 antagonist, A438079. In contrast, ATP failed to induce IL-1β release from blood not primed with LPS. ATP-induced IL-1β release was observed with LPS-primed blood from eight different pedigrees or cross breeds. Long-term incubations (24h) with LPS induced IL-1β release in canine blood in a concentration-dependent manner. This process was not altered by co-incubation with A438079. LPS-induced IL-1β release was observed with blood from 10 different pedigrees or cross breeds. These results demonstrate that both extracellular ATP and LPS can induce IL-1β release in dogs, and that ATP- but not LPS-induced IL-1β release in blood is dependent on P2X7 activation. These findings support the role of both P2X7 and TLR4 in IL-1β release in dogs.
authors.affiliation: School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.
authors.forename: Mari
authors.initials: M
authors.lastname: Spildrejorde
authors.affiliation: Albion Park Veterinary Hospital, Albion Park, NSW 2527, Australia.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Curtis
authors.affiliation: Albion Park Veterinary Hospital, Albion Park, NSW 2527, Australia.
authors.forename: Belinda L
authors.initials: BL
authors.lastname: Curtis
authors.affiliation: School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia; Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia. Electronic address: rsluyter@uow.edu.au.
authors.forename: Ronald
authors.initials: R
authors.lastname: Sluyter
mesh_terms: D000255:Adenosine Triphosphate / Q000276:immunology*; D000818:Animals; D004285:Dogs / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D053583:Interleukin-1beta / Q000097:blood / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology*; D058920:Purinergic P2X Receptor Antagonists / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D058486:Receptors, Purinergic P2X7 / Q000276:immunology*; D013777:Tetrazoles / Q000494:pharmacology
chemical_list: C523668:3-(5-(2,3-dichlorophenyl)-1H-tetrazol-1-yl)methylpyridine; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D058920:Purinergic P2X Receptor Antagonists; D011725:Pyridines; D058486:Receptors, Purinergic P2X7; D013777:Tetrazoles; D000255:Adenosine Triphosphate
keywords: ATP; Damage-associated molecular pattern receptor; Dog; ELISA; IL; Interleukin-1β; LPS; PBMCs; Pathogen-associated molecular pattern receptor; Purinergic receptor; TLR; Toll-like receptor; adenosine 5′-triphosphate; enzyme-linked immunosorbent assay; interleukin; lipopolysaccharide; peripheral blood mononuclear cells