Multi-Inhibitory Effects of A2A Adenosine Receptor Signaling on Neutrophil Adhesion Under Flow.
A2A adenosine receptor (A2AAR) signaling negatively regulates inflammatory responses in many disease models, but the detailed mechanisms remain unclear. We used the selective A2AAR agonist, ATL313, to examine how A2AAR signaling affects human and murine neutrophil adhesion under flow. Treating neutrophils with ATL313 inhibited selectin-induced, β2 integrin-dependent slow rolling and chemokine-induced, β2 integrin-dependent arrest on ICAM-1. ATL313 inhibited selectin-induced β2 integrin extension, which supports slow rolling, and chemokine-induced hybrid domain "swing-out," which supports arrest. Furthermore, ATL313 inhibited integrin outside-in signaling as revealed by reduced neutrophil superoxide production and spreading on immobilized anti-β2 integrin Ab. ATL313 suppressed selectin-triggered activation of Src family kinases (SFKs) and p38 MAPK, chemokine-triggered activation of Ras-related protein 1, and β2 integrin-triggered activation of SFKs and Vav cytoskeletal regulatory proteins. ATL313 activated protein kinase A and its substrate C-terminal Src kinase, an inhibitor of SFKs. Treating neutrophils with a protein kinase A inhibitor blocked the actions of ATL313. In vivo, ATL313-treated neutrophils rolled faster and arrested much less frequently in postcapillary venules of the murine cremaster muscle after TNF-α challenge. Furthermore, ATL313 markedly suppressed neutrophil migration into the peritoneum challenged with thioglycollate. ATL313 did not affect A2AAR-deficient neutrophils, confirming its specificity. Our findings provide new insights into the anti-inflammatory mechanisms of A2AAR signaling and the potential utility of A2AAR agonists in inflammatory diseases.
Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104;
Tadayuki
T
Yago
Immunobiology and Cancer Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; Laboratory of Oncology, Pharmacy Practice and Sciences, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan;
Hiroki
H
Tsukamoto
Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104;
Zhenghui
Z
Liu
Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104; and.
Ying
Y
Wang
Immunobiology and Cancer Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104 linda-thompson@omrf.org rodger-mcever@omrf.org.
Linda F
LF
Thompson
Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104; and linda-thompson@omrf.org rodger-mcever@omrf.org.
Rodger P
RP
McEver
D000818:Animals; D018821:CD18 Antigens / Q000235:genetics / Q000276:immunology; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D006801:Humans; D036904:Leukocyte Rolling / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D009504:Neutrophils / Q000166:cytology / Q000276:immunology*; D010880:Piperidines / Q000494:pharmacology; D051567:Proto-Oncogene Proteins c-vav / Q000235:genetics / Q000276:immunology; D043705:Receptor, Adenosine A2A / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology; D019061:src-Family Kinases / Q000235:genetics / Q000276:immunology
C510376:ATL 313; D018821:CD18 Antigens; D010880:Piperidines; D051567:Proto-Oncogene Proteins c-vav; D043705:Receptor, Adenosine A2A; D014409:Tumor Necrosis Factor-alpha; D019061:src-Family Kinases