title: Human cytomegalovirus infection induces leukotriene B4 and 5-lipoxygenase expression in human placentae and umbilical vein endothelial cells.
abstract: BACKGROUND
  Human cytomegalovirus (HCMV) can cause congenital infection with risk of neurological disability. Maternal-fetal transmission is associated with placental inflammation. 5-lipoxygenase (5-LO) is the key enzyme in the biosynthesis of Leukotrienes (LTs), which are proinflammatory mediators. This study investigated the effect of HCMV infection on 5-LO expression and Leukotriene-B4 (LTB4) induction in human placentae and umbilical vein endothelial cells (HUVEC).
  
  
  METHODS
  Seven placentae from fetuses with congenital HCMV infection and brain damage and six controls were stained with HCMV-immediate-early-antigen (HCMV-IEA) and 5-LO by immunohistochemistry. 5-hydroxyeicosatetraenoic acid (5-HETE) and LTB4 were measured in culture supernatant from ex vivo HCMV-infected placental histocultures by liquid chromatography. In vitro, HCMV infected HUVEC cells were analyzed for 5-LO mRNA and protein expression by real time PCR and immunofluorescence staining.
  
  
  RESULTS
  HCMV-IEA was abundant in all HCMV infected placentae but absent in control placentae. 5-LO expression was higher in endothelial and smooth muscle cells of HCMV-infected placentae, compared to control placentae. HCMV infection induced an up-regulation of LTB4 in ex vivo placental explants with higher levels of LTB4 at 72 h compared to controls (p = 0.002). In vitro, 5-LO transcript and protein expression were significantly induced in HCMV-infected HUVEC, compared to the control cultures (p = 0.036).
  
  
  CONCLUSIONS
  The presence of HCMV coincided with high 5-LO expression in cells of in vivo HCMV infected placentae. HCMV induced up-regulation of 5-LO in both ex vivo HCMV-infected placental explants and HUVEC. HCMV induced LT-biosynthesis in congenitally infected placentae may have a role in pathogenesis of congenital HCMV disease.
authors.affiliation: INSERM UMR 1043, Hospital Purpan, Paul Sabatier University, Toulouse 31024, France; Neonatal Unit, Children's Hospital, Toulouse 31059, France.
authors.forename: M
authors.initials: M
authors.lastname: Benard
authors.affiliation: Department of Medicine, Solna Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, SE 171 76 Stockholm, Sweden.
authors.forename: K
authors.initials: K
authors.lastname: Straat
authors.affiliation: Department of Medicine, Solna Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, SE 171 76 Stockholm, Sweden.
authors.forename: S
authors.initials: S
authors.lastname: Omarsdottir
authors.affiliation: INSERM UMR 1043, Hospital Purpan, Paul Sabatier University, Toulouse 31024, France.
authors.forename: K
authors.initials: K
authors.lastname: Leghmari
authors.affiliation: INSERM UMR U1048, I2MC, Hospital Purpan, Toulouse 31432, France.
authors.forename: J
authors.initials: J
authors.lastname: Bertrand
authors.affiliation: INSERM UMR 1043, Hospital Purpan, Paul Sabatier University, Toulouse 31024, France.
authors.forename: C
authors.initials: C
authors.lastname: Davrinche
authors.affiliation: Department of Anatomopathology, Hospital Purpan, Toulouse 31059, France.
authors.forename: I
authors.initials: I
authors.lastname: Duga-Neulat
authors.affiliation: Department of Medicine, Solna Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, SE 171 76 Stockholm, Sweden.
authors.forename: C
authors.initials: C
authors.lastname: Söderberg-Nauclér
authors.affiliation: Department of Medicine, Solna Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, SE 171 76 Stockholm, Sweden. Electronic address: afsar.rahbar@ki.se.
authors.forename: A
authors.initials: A
authors.lastname: Rahbar
authors.affiliation: INSERM UMR 1043, Hospital Purpan, Paul Sabatier University, Toulouse 31024, France; Neonatal Unit, Children's Hospital, Toulouse 31059, France; Department of Medicine, Solna Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, SE 171 76 Stockholm, Sweden. Electronic address: casper.c@chu-toulouse.fr.
authors.forename: C
authors.initials: C
authors.lastname: Casper
mesh_terms: D001094:Arachidonate 5-Lipoxygenase / Q000032:analysis* / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000151:congenital* / Q000201:enzymology / Q000378:metabolism; D042783:Endothelial Cells / Q000737:chemistry* / Q000201:enzymology; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis; D007150:Immunohistochemistry; D007975:Leukotriene B4 / Q000032:analysis*; D010920:Placenta / Q000737:chemistry* / Q000201:enzymology; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D014471:Umbilical Veins / Q000737:chemistry* / Q000201:enzymology; D015854:Up-Regulation
chemical_list: D006893:Hydroxyeicosatetraenoic Acids; D012333:RNA, Messenger; D007975:Leukotriene B4; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; D001094:Arachidonate 5-Lipoxygenase
keywords: 5-Lipoxygenase; Endothelial cells; Histoculture; Human cytomegalovirus; LTB4; Placenta