Long-term productive human cytomegalovirus infection of a human neuroblastoma cell line.
Human neuroblastoma cell line UKF-NB-4 persistently infected with human cytomegalovirus (HCMV) strain AD169 was established to study the effects of long-term HCMV infection on virus production and phenotypic characteristics of tumour cells. The cells designated UKF-NB-4AD169 were subcultured (80 subcultures) over a period of more than 2 years after initiation of infection. UKF-NB-4AD169 cells continued to produce infectious virus in successive passages, with a titre ranging from 9 x 10(3) to 1 x 10(5) and from 2 x 10(1) to 2 x 10(2) plaque-forming units per 10(6) cells and 1 ml culture medium, respectively; 10-20% of the cells produced HCMV-specific antigens, while 6-13% produced infectious virus progeny. The number of HCMV-specific DNA copies ranged from 9 x 10(4) to 9 x 10(6) per 10(6) cells. Transmission electron microscopy confirmed the productive nature of HCMV infection. UKF-NB-4AD169 cultures proliferated, with population doubling time ranging from 24.5 to 26.6 hr (19.5 to 20.3 hr for UKF-NB-4) and cell viability from 79% to 85% (91-96% for UKF-NB-4). Significantly lower amounts of tyrosine hydroxylase and decreased activity for dopamine-beta-hydroxylase than in uninfected cells were observed in UKF-NB-4AD169 cells. However, the expression of N-myc oncoprotein was significantly increased in persistently infected cultures. Our results show that long-term productive HCMV infection of UKF-NB-4 cell line is associated with the modulation of phenotypic properties, which may be related to the biological behaviour of neuroblastoma cells.
Department of Medical Virology, J.W. Goethe-University, Frankfurt a.M., Germany.
J
J
Cinatl
J U
JU
Vogel
J
J
Cinatl
B
B
Weber
H
H
Rabenau
M
M
Novak
B
B
Kornhuber
H W
HW
Doerr
D002454:Cell Differentiation; D003587:Cytomegalovirus / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000382:microbiology* / Q000473:pathology; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D006801:Humans; D009447:Neuroblastoma; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000382:microbiology*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D014779:Virus Replication
D016271:Proto-Oncogene Proteins c-myc; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase