title: Ex vivo model of congenital cytomegalovirus infection and new combination therapies.
abstract: BACKGROUND
  Congenital human cytomegalovirus (HCMV) infection is a major public health problem due to severe sequelae in the fetus and newborns. Currently, due to their toxicity anti-CMV treatments cannot be administered to pregnant women. We thus developed an ex vivo model of 1(st) trimester placental CMV infection to observe the route of infection across the placenta and to test the efficacy of various new drugs targeting different stages of viral cycle.
  
  
  METHODS
  After validation of the viability of floating villi explants by ELISA β-HCG, the kinetics of placental infection were determined by immunochemistry and qPCR in this ex vivo model. Antiviral susceptibility was determined in vitro using focus reduction assay and by qPCR in the ex vivo model.
  
  
  RESULTS
  The ex vivo model showed viral infection in trophoblasts and mesenchymal space of floating villi. In vitro, antiviral combinations of maribavir with baïcalein or artesunate inhibited viral infection by more than 90%. On the other hand, in ex vivo model, infection was reduced by 40% in presence of maribavir and artesunate. The synergistic effect observed in vitro was not observed ex vivo.
  
  
  CONCLUSIONS
  This model allowed us to understand the CMV spread in 1(st) trimester floating villi better and to analyze the anti-CMV efficacy and toxicity of new drugs that could be administered to pregnant women, either alone or in combination.
  
  
  CONCLUSIONS
  Such an ex vivo model could be applied to other viruses such as rubella or parvovirus B19 and in new drug development.
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France.
authors.forename: L
authors.initials: L
authors.lastname: Morère
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France.
authors.forename: D
authors.initials: D
authors.lastname: Andouard
authors.affiliation: CHU de Limoges, Anatomopathology Department, Limoges, France.
authors.forename: F
authors.initials: F
authors.lastname: Labrousse
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France.
authors.forename: F
authors.initials: F
authors.lastname: Saade
authors.affiliation: Faculty of Pharmacy, University of Limoges, LCSN-EA1069, Limoges, France.
authors.forename: C-A
authors.initials: CA
authors.lastname: Calliste
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France.
authors.forename: S
authors.initials: S
authors.lastname: Cotin
authors.affiliation: CHU de Limoges, Gynecology-Obstetrics Department, Limoges, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Aubard
authors.affiliation: Virology Division, SEALS Microbiology Prince of Wales Hospital and SOMS, BABS UNSW, Randwick, Australia.
authors.forename: W D
authors.initials: WD
authors.lastname: Rawlinson
authors.affiliation: CHU de Limoges, Mother and Child Hospital, Histopathology Department, Limoges, France.
authors.forename: F
authors.initials: F
authors.lastname: Esclaire
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France.
authors.forename: S
authors.initials: S
authors.lastname: Hantz
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, Limoges, France.
authors.forename: M-C
authors.initials: MC
authors.lastname: Ploy
authors.affiliation: University of Limoges, Inserm U1092, Limoges, France; INSERM, UMR 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, Limoges, France; French National Reference Center for Cytomegaloviruses, Limoges, France. Electronic address: sophie.alain@unilim.fr.
authors.forename: S
authors.initials: S
authors.lastname: Alain
mesh_terms: D000328:Adult; D000998:Antiviral Agents / Q000494:pharmacology / Q000627:therapeutic use*; D037621:Artemisinins / Q000494:pharmacology / Q000627:therapeutic use; D000077332:Artesunate; D001562:Benzimidazoles / Q000494:pharmacology / Q000627:therapeutic use; D003586:Cytomegalovirus Infections / Q000151:congenital* / Q000188:drug therapy*; D005260:Female; D044950:Flavanones / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000188:drug therapy* / Q000821:virology; D012263:Ribonucleosides / Q000494:pharmacology / Q000627:therapeutic use; D014327:Trophoblasts / Q000187:drug effects / Q000821:virology*; D004004:Dichlororibofuranosylbenzimidazole / Q000031:analogs & derivatives
chemical_list: D000998:Antiviral Agents; D037621:Artemisinins; D000077332:Artesunate; D001562:Benzimidazoles; D044950:Flavanones; D012263:Ribonucleosides; C006680:baicalein; C400401:maribavir; D004004:Dichlororibofuranosylbenzimidazole
keywords: Artesunate; Ex vivo model; Flavonoïds; Maribavir; Placental infection