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.
J Fish Dis
; 38(6): 507-14, 2015 Jun.
Article
en En
| MEDLINE
| ID: mdl-24820532
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.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Temperatura
/
Línea Celular
/
Latencia del Virus
/
Infecciones por Herpesviridae
/
Enfermedades de los Peces
/
Herpesviridae
Límite:
Animals
Idioma:
En
Revista:
J Fish Dis
Asunto de la revista:
BIOLOGIA
/
MEDICINA VETERINARIA
Año:
2015
Tipo del documento:
Article
País de afiliación:
Japón