Concomitant activation of P2Y(2) and P2Y(6) receptors on monocytes is required for TLR1/2-induced neutrophil migration by regulating IL-8 secretion.
Extracellular nucleotides regulate a variety of cellular responses involved in inflammation via the activation of P2 receptors. Here, we show that nucleotides regulate TLR2-induced neutrophil migration both in vivo and in vitro. The nucleotide scavenger apyrase inhibited neutrophil recruitment in murine air pouches injected with the TLR2 agonist Pam(3)CSK(4). In agreement, the supernatants of either human primary monocytes or monocytic cells (THP-1 and U937) treated with Pam(3)CSK(4) recruited significantly fewer neutrophils when the former cells were treated in the presence of apyrase. As demonstrated with inhibitory Ab, these supernatants induced neutrophil migration due to IL-8 secretion. In addition, IL-8 secretion was markedly diminished by the non-selective P2 receptor antagonists reactive blue 2 and suramin, and by a selective P2Y(6) antagonist, MRS2578. Selective antagonists of P2Y(1) (MRS2500) and P2Y(11) (NF157) did not affect IL-8 release. The knockdown of either P2Y(2) or P2Y(6) with specific shRNA diminished IL-8 secretion from Pam(3)CSK(4)-treated THP-1 cells. Altogether, these results show that extracellular nucleotides, via P2Y(2) and P2Y(6) receptors, regulate neutrophil migration by controlling TLR2-induced IL-8 release from human monocytes. In line with our previous work on TLR4, this study further supports the importance of nucleotides in bacterial-induced neutrophil migration.
Ben Yebdri
Fethia
F
Centre de Recherche en Rhumatologie et Immunologie, Centre Hospitalier Universitaire de Québec, Université Laval, Québec, QC, Canada.
Kukulski
Filip
F
Tremblay
Alain
A
Sévigny
Jean
J
D000243:Adenosine Deaminase / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D001081:Apyrase / Q000494:pharmacology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005260:Female; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism*; D055666:Lipopeptides / Q000494:pharmacology; D051379:Mice; D008815:Mice, Inbred Strains; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism*; D009504:Neutrophils / Q000166:cytology*; D009711:Nucleotides / Q000378:metabolism / Q000494:pharmacology; D058910:Purinergic P2 Receptor Agonists; D058919:Purinergic P2 Receptor Antagonists; D034622:RNA Interference; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D058473:Receptors, Purinergic P2Y2; D051194:Toll-Like Receptor 1 / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000819:agonists / Q000378:metabolism*; D051198:Toll-Like Receptor 5 / Q000819:agonists; D051200:Toll-Like Receptor 8 / Q000819:agonists; D020298:U937 Cells
D000906:Antibodies; D017077:Culture Media, Conditioned; D016209:Interleukin-8; D055666:Lipopeptides; D009711:Nucleotides; C546385:P2RY2 protein, human; C546386:P2ry2 protein, mouse; C508075:Pam(3)CSK(4) peptide; D058910:Purinergic P2 Receptor Agonists; D058919:Purinergic P2 Receptor Antagonists; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; C495346:TLR5 protein, human; C495349:TLR8 protein, human; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2; D051198:Toll-Like Receptor 5; D051200:Toll-Like Receptor 8; C099845:purinoceptor P2Y6; D000243:Adenosine Deaminase; D001081:Apyrase