Activation of the damage-associated molecular pattern receptor P2X7 induces interleukin-1β release from canine monocytes.
P2X7, a damage-associated molecular pattern receptor and adenosine 5'-triphosphate (ATP)-gated cation channel, plays an important role in the activation of the NALP3 inflammasome and subsequent release of interleukin (IL)-1β from human monocytes; however its role in monocytes from other species including the dog remains poorly defined. This study investigated the role of P2X7 in canine monocytes, including its role in IL-1β release. A fixed-time flow cytometric assay demonstrated that activation of P2X7 by extracellular ATP induces the uptake of the organic cation, YO-PRO-1(2+), into peripheral blood monocytes from various dog breeds, a process impaired by the specific P2X7 antagonist, A438079. Moreover, in five different breeds, relative P2X7 function in monocytes was about half that of peripheral blood T cells but similar to that of peripheral blood B cells. Reverse transcription-PCR demonstrated the presence of P2X7, NALP3, caspase-1 and IL-1β in LPS-primed canine monocytes. Immunoblotting confirmed the presence of P2X7 in LPS-primed canine monocytes. Finally, extracellular ATP induced YO-PRO-1(2+) uptake into and IL-1β release from these cells, with both processes impaired by A438079. These results demonstrate that P2X7 activation induces the uptake of organic cations into and the release of IL-1β from canine monocytes. These findings indicate that P2X7 may play an important role in IL-1β-dependent processes in dogs.
School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia.
Iman
I
Jalilian
Michelle
M
Peranec
Belinda L
BL
Curtis
Aine
A
Seavers
Mari
M
Spildrejorde
Vanessa
V
Sluyter
Ronald
R
Sluyter
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001583:Benzoxazoles / Q000276:immunology; D002460:Cell Line; D004285:Dogs / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D015151:Immunoblotting; D058847:Inflammasomes / Q000037:antagonists & inhibitors / Q000276:immunology; D053583:Interleukin-1beta / Q000235:genetics / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D051379:Mice; D009000:Monocytes / Q000276:immunology*; D011725:Pyridines / Q000494:pharmacology; D011806:Quinolinium Compounds / Q000276:immunology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013777:Tetrazoles / Q000494:pharmacology
C523668:3-(5-(2,3-dichlorophenyl)-1H-tetrazol-1-yl)methylpyridine; D001583:Benzoxazoles; D058847:Inflammasomes; D053583:Interleukin-1beta; D011725:Pyridines; D011806:Quinolinium Compounds; D058486:Receptors, Purinergic P2X7; D013777:Tetrazoles; C089813:YO-PRO 1; D012313:RNA; D000255:Adenosine Triphosphate