title: Establishment of a new cell line susceptible to Cyprinid herpesvirus 3 (CyHV-3) and possible latency of CyHV-3 by temperature shift in the cells.
abstract: A new cell line named CCF-K104 predominantly consisting of fibroblastic cells showed optimal growth at temperatures from 25 °C to 30 °C. Serial morphological changes in the cells induced by Cyprinid herpesvirus 3 (CyHV-3) included cytoplasmic vacuolar formation, cell rounding and detachment. Mature virions were purified from CyHV-3-infected CCF-K104 cells by sucrose gradient ultracentrifugation and had a typical herpesvirus structure on electron microscopy. Infectious CyHV-3 was produced stably in CCF-K104 cells over 30 viral passages. Our findings showed that CCF-K104 is a useful cell line for isolation and productive replication of CyHV-3. A temperature shift from 25 °C to 15 °C or 35 °C did not allow serial morphological changes as observed at 25 °C for 14 days. Under the same conditions, real-time PCR showed that CyHV-3 was present with low viral DNA loads, suggesting that CyHV-3 may establish latent infection in CCF-K104 cells. Amplification of the left and right terminal repeat sequences of the CyHV-3 genome arranged in a head-to-tail manner was detected by nested PCR following an upshift in temperature from 25 °C to 35 °C. The PCR results suggested that the circular genome may represent a latent form of CyHV-3.
authors.affiliation: Laboratory of Fish Disease, Faculty of Agriculture, Kochi University, Nankoku, Kochi, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Imajoh
authors.forename: H
authors.initials: H
authors.lastname: Fujioka
authors.forename: K
authors.initials: K
authors.lastname: Furusawa
authors.forename: K
authors.initials: K
authors.lastname: Tamura
authors.forename: K
authors.initials: K
authors.lastname: Yamasaki
authors.forename: S
authors.initials: S
authors.lastname: Kurihara
authors.forename: J
authors.initials: J
authors.lastname: Yamane
authors.forename: K
authors.initials: K
authors.lastname: Kawai
authors.forename: S
authors.initials: S
authors.lastname: Oshima
mesh_terms: D000818:Animals; D002347:Carps; D002460:Cell Line; D005393:Fish Diseases / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics; D006564:Herpesviridae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology* / Q000648:ultrastructure; D006566:Herpesviridae Infections / Q000662:veterinary* / Q000821:virology; D008969:Molecular Sequence Data; D013696:Temperature; D017735:Virus Latency / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology
keywords: Cyprinid herpesvirus 3; caudal fin cell line; common carp; latency; temperature shift; terminal direct repeat