title: Human cytomegalovirus chemokine receptor US28 induces migration of cells on a CX3CL1-presenting surface.
abstract: Human cytomegalovirus (HCMV)-encoded G protein-coupled-receptor US28 is believed to participate in virus dissemination through modulation of cell migration and immune evasion. US28 binds different CC chemokines and the CX3C chemokine CX3CL1. Membrane-anchored CX3CL1 is expressed by immune-activated endothelial cells, causing redirection of CX3CR1-expressing leukocytes in the blood to sites of infection. Here, we used stable transfected cell lines to examine how US28 expression affects cell migration on immobilized full-length CX3CL1, to model how HCMV-infected leukocytes interact with inflamed endothelium. We observed that US28-expressing cells migrated more than CX3CR1-expressing cells when adhering to immobilized CX3CL1. US28-induced migration was G protein-signalling dependent and was blocked by the phospholipase Cβ inhibitor U73122 and the intracellular calcium chelator BAPTA-AM. In addition, migration was inhibited in a dose-dependent manner by competition from CCL2 and CCL5, whereas CCL3 had little effect. Instead of migrating, CX3CR1-expressing cells performed 'dancing-on-the-spot' movements, demonstrating that anchored CX3CL1 acts as a strong tether for these cells. At low receptor expression levels, however, no significant difference in migration potential was observed when comparing the migration of CX3CR1- and US28-expressing cells. Thus, these data showed that, in contrast to CX3CR1, which promotes efficient cell capture upon binding to anchored CX3CL1, US28 acts to increase the migration of cells upon binding to the same ligand. Overall, this indicates that infected cells probably move more than uninfected cells in inflamed tissues with high CX3CL1 expression, with soluble chemokines affecting the final migration.
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech, DK-2800 Kgs Lyngby, Denmark.
authors.forename: Gertrud M
authors.initials: GM
authors.lastname: Hjortø
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech, DK-2800 Kgs Lyngby, Denmark.
authors.forename: Katrine
authors.initials: K
authors.lastname: Kiilerich-Pedersen
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech, DK-2800 Kgs Lyngby, Denmark.
authors.forename: David
authors.initials: D
authors.lastname: Selmeczi
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech, DK-2800 Kgs Lyngby, Denmark.
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Kledal
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech, DK-2800 Kgs Lyngby, Denmark.
authors.forename: Niels B
authors.initials: NB
authors.lastname: Larsen
mesh_terms: D000073883:CX3C Chemokine Receptor 1; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D054428:Chemokine CX3CL1 / Q000235:genetics / Q000378:metabolism*; D019742:Chemokines, CC / Q000378:metabolism; D003587:Cytomegalovirus / Q000378:metabolism*; D003586:Cytomegalovirus Infections / Q000821:virology*; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D042783:Endothelial Cells; D004963:Estrenes / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D054799:Phospholipase C beta / Q000037:antagonists & inhibitors; D011760:Pyrrolidinones / Q000494:pharmacology; D019707:Receptors, Chemokine / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D059008:Time-Lapse Imaging; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000073883:CX3C Chemokine Receptor 1; C095235:CX3CR1 protein, human; D002614:Chelating Agents; D054428:Chemokine CX3CL1; D019742:Chemokines, CC; D004963:Estrenes; D008565:Membrane Proteins; D010726:Phosphodiesterase Inhibitors; D011760:Pyrrolidinones; D019707:Receptors, Chemokine; C411191:US28 receptor, Cytomegalovirus; D014764:Viral Proteins; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D054799:Phospholipase C beta; D019204:GTP-Binding Proteins