title: Cholesterol-rich lipid rafts play an important role in the Cyprinid herpesvirus 3 replication cycle.
abstract: The Cyprinus herpesvirus 3 (CyHV-3) is a member of the new Alloherpesviridae virus family in the Herpesvirales order. CyHV-3 has been implicated in a large number of disease outbreaks in carp populations causing up to 100% mortality. The aim of this study was to investigate the requirement of cholesterol-rich lipid rafts in CyHV-3 entry and replication in carp cells. Plasma membrane cholesterol was depleted from common carp brain (CCB) cells with methyl-β-cyclodextrin (MβCD). Treated and non-treated cells were infected with CyHV-3 and virus binding and infection parameters were assessed using RT-qPCR, immunocytochemistry and virus titration. The effect of cholesterol reduction severely stunted virus entry in vitro, however after cholesterol replenishment virus entry and subsequent replication rates were similar to the control infection. Furthermore, cholesterol depletion did not significantly influence virus binding and the subsequent post-entry replication stage, however had an impact on virus egress. Comparative analysis of the lipid compositions of CyHV-3 and CCB membrane fractions revealed strong similarities between the lipid composition of the CyHV-3 and CCB lipid rafts. The results presented here show that cholesterol-rich lipid rafts are important for the CyHV-3 replication cycle especially during entry and egress.
authors.affiliation: Fish Disease Research Unit, University of Veterinary Medicine Hanover, Germany.
authors.forename: Graham
authors.initials: G
authors.lastname: Brogden
authors.affiliation: Fish Disease Research Unit, University of Veterinary Medicine Hanover, Germany.
authors.forename: Mikołaj
authors.initials: M
authors.lastname: Adamek
authors.affiliation: Fish Disease Research Unit, University of Veterinary Medicine Hanover, Germany.
authors.forename: Marcus J
authors.initials: MJ
authors.lastname: Proepsting
authors.affiliation: Department of Pathology, University of Veterinary Medicine Hanover, Germany.
authors.forename: Reiner
authors.initials: R
authors.lastname: Ulrich
authors.affiliation: Department of Physiological Chemistry, University of Veterinary Medicine Hanover, Germany.
authors.forename: Hassan Y
authors.initials: HY
authors.lastname: Naim
authors.affiliation: Fish Disease Research Unit, University of Veterinary Medicine Hanover, Germany. Electronic address: Dieter.steinhagen@tiho-hannover.de.
authors.forename: Dieter
authors.initials: D
authors.lastname: Steinhagen
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology; D002347:Carps / Q000821:virology*; D002784:Cholesterol / Q000378:metabolism*; D006564:Herpesviridae / Q000502:physiology*; D021962:Membrane Microdomains / Q000378:metabolism / Q000502:physiology*; D053585:Virus Attachment; D053586:Virus Internalization; D014779:Virus Replication / Q000502:physiology*; D047392:beta-Cyclodextrins
chemical_list: D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002784:Cholesterol