title: Cutthroat Trout Virus-Towards a Virus Model to Support Hepatitis E Research.
abstract: Cutthroat trout virus (CTV) is a non-pathogenic fish virus belonging to theHepeviridaefamily, and it is distantly related to hepatitis E virus (HEV). Here, we report the development of an efficient cell culture system where CTV can consistently replicate to titers never observed before with a hepevirus. By using the rainbow trout gill (RTGill-W1) cell line, CTV reaches 1010geq/mL intracellularly and 10⁸ geq/mL extracellularly within 5-6 days in culture. We additionally established a qPCR system to investigate CTV infectivity, and developed a specific antibody directed against the viral capsid protein encoded by ORF2. With these methods, we were able to follow the progressive accumulation of viral RNA and the capsid protein, and their intracellular distribution during virus replication. Virus progeny purified through iodixanol density gradients indicated-that similar to HEV-CTV produced in cell culture is also lipid-associated. The lack of an efficient cell culture system has greatly impeded studies with HEV, a major human pathogen that causes hepatitis worldwide. Although several cell culture systems have recently been established, the replication efficiency of HEV is not robust enough to allow studies on different aspects of the virus replication cycle. Therefore, a surrogate virus that can replicate easily and efficiently in cultured cells would be helpful to boost research studies with hepeviruses. Due to its similarities, but also its key differences to HEV, CTV represents a promising tool to elucidate aspects of the replication cycle ofHepeviridaein general, and HEV in particular.
authors: Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. marcus.vonnordheim@dcb.unibe.ch.
authors: Marcus
authors: M
authors: von Nordheim
authors: Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. michel.boinay@students.unibe.ch.
authors: Michel
authors: M
authors: Boinay
authors: Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. remo.leisi@dcb.unibe.ch.
authors: Remo
authors: R
authors: Leisi
authors: Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. christoph.kempf@dcb.unibe.ch.|CSL Behring AG, Bern 3014, Switzerland. christoph.kempf@dcb.unibe.ch.
authors: Christoph
authors: C
authors: Kempf
authors: Department of Chemistry and Biochemistry, University of Bern, Bern 3012, Switzerland. carlos.ros@dcb.unibe.ch.|CSL Behring AG, Bern 3014, Switzerland. carlos.ros@dcb.unibe.ch.
authors: Carlos
authors: C
authors: Ros
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D002460:Cell Line; D052678:Hepevirus / Q000502:physiology*; D007118:Immunoassay / Q000379:methods; D017686:Oncorhynchus mykiss; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D014776:Virus Cultivation / Q000379:methods*; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D012367:RNA, Viral
keywords: cutthroat trout virus; hepatitis E virus; virus model