title: Endothelial P2Y2 receptor regulates LPS-induced neutrophil transendothelial migration in vitro.
abstract: Previous studies showed that P2 receptors are involved in neutrophil migration via stimulation of chemokine release and by facilitating chemoattractant gradient sensing. Here, we have investigated whether these receptors are involved in LPS-induced neutrophil transendothelial migration (TEM) using a Boyden chamber where neutrophils migrated through a layer of lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs). In line with a role of P2 receptors, neutrophil TEM was inhibited by the P2 receptor antagonists suramin and reactive blue 2 (RB-2) acting on the basolateral, but not luminal, HUVECs' P2 receptors. HUVECs express P2Y(1), P2Y(2), P2Y(4), P2Y(6) and P2Y(11). The involvement of P2Y(4) was unlikely as this receptor is insensitive to suramin while P2Y(1), P2Y(6) and P2Y(11) were excluded with available selective antagonists, leaving P2Y(2) as the only candidate. Indeed, the P2Y(2) knockdown in HUVECs inhibited neutrophil TEM compared to control HUVECs transfected with scrambled siRNA. Moreover, UTP, a P2Y(2) ligand, markedly potentiated LPS-induced TEM. Interestingly, IL-8 and ICAM-1 had a modest effect on neutrophil TEM in this 3h assay which was significantly diminished by the inhibition of Rho kinase in HUVECs with Y27632. In summary, endothelial P2Y(2) receptors control the early LPS-induced neutrophil TEM in vitro via Rho kinase activation.
authors.affiliation: Centre de Recherche en Rhumatologie et Immunologie, Centre Hospitalier Universitaire de Québec, Université Laval, Québec, QC, Canada.
authors.forename: Filip
authors.initials: F
authors.lastname: Kukulski
authors.forename: Fethia
authors.initials: F
authors.lastname: Ben Yebdri
authors.forename: Fariborz
authors.initials: F
authors.lastname: Bahrami
authors.forename: Michel
authors.initials: M
authors.lastname: Fausther
authors.forename: Alain
authors.initials: A
authors.lastname: Tremblay
authors.forename: Jean
authors.initials: J
authors.lastname: Sévigny
mesh_terms: D000577:Amides / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects* / Q000235:genetics / Q000276:immunology*; D042783:Endothelial Cells / Q000276:immunology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D055785:Gene Knockdown Techniques; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D016209:Interleukin-8 / Q000235:genetics / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009504:Neutrophils / Q000276:immunology*; D058919:Purinergic P2 Receptor Antagonists; D011725:Pyridines / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000276:immunology*; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D013498:Suramin / Q000494:pharmacology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology
chemical_list: D000577:Amides; D000970:Antineoplastic Agents; C508606:CXCL8 protein, human; D004791:Enzyme Inhibitors; D016209:Interleukin-8; D008070:Lipopolysaccharides; C546382:P2RY1 protein, human; C546385:P2RY2 protein, human; D058919:Purinergic P2 Receptor Antagonists; D011725:Pyridines; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; C104749:purinoceptor P2Y4; C099845:purinoceptor P2Y6; D018799:Intercellular Adhesion Molecule-1; C108830:Y 27632; D013498:Suramin; D054460:rho-Associated Kinases