title: Comprehensive analysis of chemokine-induced cAMP-inhibitory responses using a real-time luminescent biosensor.
abstract: Chemokine receptors are members of the G-protein-coupled receptor (GPCR) family coupled to members of the Gi class, whose primary function is to inhibit the cellular adenylate cyclase. We used a cAMP-related and PKA-based luminescent biosensor (GloSensor™ F-22) to monitor the real-time downstream response of chemokine receptors, especially CX3CR1 and CXCR4, after activation with their cognate ligands CX3CL1 and CXCL12. We found that the amplitudes and kinetic profiles of the chemokine responses were conserved in various cell types and were independent of the nature and concentration of the molecules used for cAMP prestimulation, including either the adenylate cyclase activator forskolin or ligands mediating Gs-mediated responses like prostaglandin E2 or beta-adrenergic agonist. We conclude that the cAMP chemokine response is robustly conserved in various inflammatory conditions. Moreover, the cAMP-related luminescent biosensor appears as a valuable tool to analyze the details of Gi-mediated cAMP-inhibitory cellular responses, even in native conditions and could help to decipher their precise role in cell function.
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, Inserm U 1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses, 91 Boulevard de l'Hôpital, F-75013 Paris, France.
authors.forename: Virginia
authors.initials: V
authors.lastname: Felouzis
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, Inserm U 1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses, 91 Boulevard de l'Hôpital, F-75013 Paris, France.
authors.forename: Patricia
authors.initials: P
authors.lastname: Hermand
authors.affiliation: Université de Cergy-Pontoise, CNRS, UMR 8089, Laboratoire de Physique Théorique et Modélisation, 2 Avenue A. Chauvin, F-95302 Cergy-Pontoise, France.
authors.forename: Guy Trambly
authors.initials: GT
authors.lastname: de Laissardière
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, Inserm U 1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses, 91 Boulevard de l'Hôpital, F-75013 Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Combadière
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, Inserm U 1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses, 91 Boulevard de l'Hôpital, F-75013 Paris, France. Electronic address: philippe.deterre@upmc.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Deterre
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D015374:Biosensing Techniques; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D007700:Kinetics; D019718:Receptors, CXCR4 / Q000187:drug effects
chemical_list: D018925:Chemokines; D019718:Receptors, CXCR4; D005576:Colforsin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D000262:Adenylyl Cyclases; D007545:Isoproterenol
keywords: Adenylate cyclase; Biosensor; Chemokine; Forskolin; Gi; cAMP