title: Direct infection of primary endothelial cells  with human cytomegalovirus prevents  angiogenesis and migration.
abstract: Human cytomegalovirus (hCMV) is a beta herpesvirus that establishes lifelong infection. Although the virus does not usually cause overt clinical symptoms in immunocompetent individuals it can have deleterious effects in immunocompromised patients, such as those on post-transplant medication or with HIV infection. hCMV is the most common congenital infection and can lead to serious fetal sequelae. Endothelial cells (ECs) are natural hosts for hCMV in vivo, therefore, investigations of how this cell type is modulated by infection are key to understanding hCMV pathogenesis. Previous studies have examined the effect of secretomes from hCMV-infected cells on EC angiogenesis, whereas the effect of direct infection on this process has not been so well investigated. Here, we show that placental ECs are viral targets during congenital infection and that vessels in infected tissue appear morphologically abnormal. We demonstrate that the clinical hCMV strain VR1814 impaired EC tube assembly in in vitro angiogenesis assays and inhibited wound healing ability in scratch assays. Secretomes from infected cultures did not impair angiogenesis of uninfected ECs, suggesting that cell-intrinsic changes, as opposed to secreted factors, were responsible. We observed viral gene transcription dependent downregulation of the expression of angiogenesis-associated genes, including angiopoietin-2, TEK receptor and vascular endothelial growth factor receptors. An alternative clinical hCMV stain, TB40E showed similar effects on EC angiogenesis. Together, our data indicate that direct infection with hCMV can induce an anti-migratory and anti-angiogenic EC phenotype, which could have a detrimental effect on the vasculature development in infected tissues.
authors.affiliation: Clinical Chemistry, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.|Institute for Clinical Chemistry and Laboratory Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Rasmus K L
authors.initials: RKL
authors.lastname: Gustafsson
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Hannah C
authors.initials: HC
authors.lastname: Jeffery
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Koon-Chu
authors.initials: KC
authors.lastname: Yaiw
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Wilhelmi
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Ourania N
authors.initials: ON
authors.lastname: Kostopoulou
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Belghis
authors.initials: B
authors.lastname: Davoudi
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Afsar
authors.initials: A
authors.lastname: Rahbar
authors.affiliation: INSERM UMR 1043, Hospital Purpan, Paul Sabatier University, Toulouse 31024, France.
authors.forename: Melinda
authors.initials: M
authors.lastname: Benard
authors.affiliation: Clinical Chemistry, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.|Institute for Clinical Chemistry and Laboratory Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Renné
authors.affiliation: Department of Medicine, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Söderberg-Nauclér
authors.affiliation: Clinical Chemistry, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.|Institute for Clinical Chemistry and Laboratory Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Lynn M
authors.initials: LM
authors.lastname: Butler
mesh_terms: D002465:Cell Movement; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000151:congenital* / Q000821:virology; D042783:Endothelial Cells / Q000502:physiology* / Q000821:virology*; D005260:Female; D015967:Gene Expression Regulation, Viral; D006801:Humans; D018445:Infectious Disease Transmission, Vertical; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D018919:Neovascularization, Physiologic; D010920:Placenta / Q000098:blood supply / Q000166:cytology / Q000821:virology; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D016753:Interleukin-10