title: Human cytomegalovirus infection impairs endothelial cell chemotaxis by disturbing VEGF signalling and actin polymerization.
abstract: OBJECTIVE
  Human cytomegalovirus (HCMV) infection has been linked to the pathogenesis of vasculopathies; however, its pathogenic relevance remains to be established. A prerequisite for vascular repair is endothelial cell migration. We evaluated the influence of HCMV on chemokinesis and chemotactic response of human coronary artery endothelial cells (HCAEC) towards vascular endothelial growth factor (VEGF).
  
  
  RESULTS
  A virus dose-dependent reduction in chemokinesis and VEGF-dependent chemotaxis was observed (P < 0.05). UV-inactivated virus did not inhibit chemotaxis or chemokinesis, indicating that viral gene expression is mandatory. We identified two HCMV-induced mechanisms explaining the reduction of chemotaxis: first, a non-ambiguous reduction of VEGFR-2 protein was observed, due to decreased transcription. This protein down-modulation could not be inhibited by Ganciclovir. The remaining VEGFR-2 expressed on infected HCAEC was able to stimulate cell activation. Second, HCMV infection influences actin polymerization in HCAEC as shown by FACS analysis: actin polymerization was significantly reduced to 53 and 51% (P < 0.05) compared with non-infected HCAEC at 24 and 72 h p.i., respectively. Genetically and pharmacologically eliminated VEGFR-2 function resulted in a significant (P < 0.05) reduction of VEGF-induced activation of actin polymerization.
  
  
  CONCLUSIONS
  We demonstrated a significant reduction of the chemotactic mobility of HCMV-infected HCAEC mediated by down-modulation of the VEGFR-2 and by inhibition of actin polymerization. This VEGF resistance of HCMV-infected endothelial cells is likely to promote atherogenesis.
authors.affiliation: Department of Virology, Ulm University Medical Center, Ulm D-89081, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Reinhardt
authors.affiliation: Cells-in-Motion Cluster of Excellence (EXC 1003-CiM), University of Münster, Münster, Germany Department of Cardiovascular Medicine, University Hospital Muenster, Albert-Schweitzer-Campus 1-A1, Muenster D-48149, Germany.
authors.forename: Rinesh
authors.initials: R
authors.lastname: Godfrey
authors.affiliation: Department of Cardiology, University Medical Center, Ulm, Germany.
authors.forename: Guido
authors.initials: G
authors.lastname: Fellbrich
authors.affiliation: Department of Cardiology, University Medical Center, Ulm, Germany.
authors.forename: Hedwig
authors.initials: H
authors.lastname: Frank
authors.affiliation: Department of Virology, Ulm University Medical Center, Ulm D-89081, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Lüske
authors.affiliation: Department of Cardiology, Maastricht University Medical Center (MUMC) and Cardiovascular Research Institute Maastricht (CARIM), Maastricht, The Netherlands.
authors.forename: Servé
authors.initials: S
authors.lastname: Olieslagers
authors.affiliation: Department of Virology, Ulm University Medical Center, Ulm D-89081, Germany johannes.waltenberger@ukmuenster.de thomas.mertens@uniklinik-ulm.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Mertens
authors.affiliation: Cells-in-Motion Cluster of Excellence (EXC 1003-CiM), University of Münster, Münster, Germany Department of Cardiovascular Medicine, University Hospital Muenster, Albert-Schweitzer-Campus 1-A1, Muenster D-48149, Germany Department of Cardiology, University Medical Center, Ulm, Germany Department of Cardiology, Maastricht University Medical Center (MUMC) and Cardiovascular Research Institute Maastricht (CARIM), Maastricht, The Netherlands johannes.waltenberger@ukmuenster.de thomas.mertens@uniklinik-ulm.de.
authors.forename: Johannes
authors.initials: J
authors.lastname: Waltenberger
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism / Q000821:virology*; D002460:Cell Line; D002633:Chemotaxis / Q000187:drug effects*; D003587:Cytomegalovirus / Q000235:genetics / Q000472:pathogenicity*; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D054884:Host-Pathogen Interactions; D006801:Humans; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects*; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D042461:Vascular Endothelial Growth Factor A; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Chemotaxis; Endothelial cell; Human cytomegalovirus; Signal transduction; Vascular endothelial growth factor