title: Entry of bovine viral diarrhea virus into ovine cells occurs through clathrin-dependent endocytosis and low pH-dependent fusion.
abstract: Although mechanisms of bovine viral diarrhea virus (BVDV) entry into bovine cells have been elucidated, little is known concerning pestivirus entry and receptor usage in ovine cells. In this study, we determined the entry mechanisms of BVDV-1 and BVDV-2 in sheep fetal thymus cells. Both BVDV-1 and BVDV-2 infections were inhibited completely by chlorpromazine, beta-methyl cyclodextrin, sucrose, bafilomycin A1, chloroquine, and ammonium chloride. Simultaneous presence of reducing agent and low pH resulted in marked loss of BVDV infectivity. Moreover, BVDV was unable to fuse with ovine cell membrane by the presence of reducing agent or low pH alone, while combination of both led to fusion at low efficiency. Furthermore, sheep fetal thymus cells acutely infected with BVDV-1 or BVDV-2 were found protected from heterologous BVDV infection. Taken together, our results showed for the first time that entry of both BVDV-1 and BVDV-2 into ovine cells occurred through clathrin-dependent endocytosis, endosomal acidification, and low pH-dependent fusion following an activation step, besides suggesting the involvement of a common ovine cellular receptor during attachment and entry.
authors.affiliation: High Security Animal Disease Laboratory, Indian Veterinary Research Institute, Anand Nagar, Bhopal, 462021, Madhya Pradesh, India.
authors.forename: Basavaraj Shrishail
authors.initials: BS
authors.lastname: Mathapati
authors.forename: Niranjan
authors.initials: N
authors.lastname: Mishra
authors.forename: Katherukamem
authors.initials: K
authors.lastname: Rajukumar
authors.forename: Ram Kumar
authors.initials: RK
authors.lastname: Nema
authors.forename: Sthita Pragnya
authors.initials: SP
authors.lastname: Behera
authors.forename: Shiv Chandra
authors.initials: SC
authors.lastname: Dubey
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D002746:Chlorpromazine / Q000494:pharmacology; D002966:Clathrin / Q000502:physiology*; D030241:Diarrhea Virus 1, Bovine Viral / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology; D030242:Diarrhea Virus 2, Bovine Viral / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology; D001908:Diarrhea Viruses, Bovine Viral / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D012756:Sheep; D013395:Sucrose / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology; D053586:Virus Internalization / Q000187:drug effects; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D002966:Clathrin; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D000643:Ammonium Chloride; D013395:Sucrose; D002738:Chloroquine; D002746:Chlorpromazine