TY - JOUR

T1 - Human cytomegalovirus chemokine receptor US28 induces migration of cells on a CX3CL1-presenting surface

AU - Hjortø, Gertrud M

AU - Kiilerich-Pedersen, Katrine

AU - Selmeczi, David

AU - Kledal, Thomas N.

AU - Larsen, Niels B

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

KW - Cell Movement

KW - Cells, Cultured

KW - Chelating Agents

KW - Chemokine CX3CL1

KW - Chemokines, CC

KW - Cytomegalovirus

KW - Cytomegalovirus Infections

KW - Dose-Response Relationship, Drug

KW - Egtazic Acid

KW - Endothelial Cells

KW - Estrenes

KW - GTP-Binding Proteins

KW - HEK293 Cells

KW - Humans

KW - Membrane Proteins

KW - Mutation

KW - Phosphodiesterase Inhibitors

KW - Phospholipase C beta

KW - Pyrrolidinones

KW - Receptors, Chemokine

KW - Signal Transduction

KW - Time-Lapse Imaging

KW - Viral Proteins

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1099/vir.0.047290-0

DO - 10.1099/vir.0.047290-0

M3 - Journal article

C2 - 23303826

VL - 94

SP - 1111

EP - 1120

JO - Journal of General Virology

JF - Journal of General Virology

SN - 0022-1317

IS - Pt 5

ER -