Effect of enforced expression of human bcl-2 on Japanese encephalitis virus-induced apoptosis in cultured cells.
Infection by Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, causes acute encephalitis in humans and induces severe cytopathic effects in different types of cultured cells. This study attempted to determine whether apoptosis contributes to virus-induced cell death in a culture system by characterizing JEV lytic infection in baby hamster kidney BHK-21 cells, murine neuroblastoma N18 cells, and human neuronal progenitor NT2 cells. According to our results, the replication of JEV, and not the UV-inactivated virions per se, triggered apoptosis in these cell lines, as evidenced by nuclear condensation, DNA fragmentation ladder, and in situ end labeling of DNA strand breaks with terminal transferase (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling assay). Different strains of JEV, regardless of whether they are neurovirulent to mice, could induce apoptosis of the infected cells. In addition, enforced expression of the human protooncogene bcl-2 in BHK-21 cells, which did not influence virus production, appeared to delay the process of JEV-induced apoptosis, despite the fact that most infected cells were inevitably killed after prolonged cultures. However, Bcl-2 proteins expressed in N18 cells failed to block JEV-induced apoptosis, although they did prevent Sindbis virus-induced apoptosis from occurring in the same cells. This finding suggests that these two viruses may utilize similar but not identical mechanisms to kill their infected cells. The results presented here thus demonstrate that apoptosis can be a general mechanism for JEV-induced cell death and that enforced bcl-2 expression may be inadequate in protecting all cell types from JEV-induced apoptosis in cell cultures.
Liao
C L
CL
Institute of Preventive Medicine, Department of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, Republic of China. chinglen@ndmc1.ndmctsgh.edu.tw
Lin
Y L
YL
Wang
J J
JJ
Huang
Y L
YL
Yeh
C T
CT
Ma
S H
SH
Chen
L K
LK
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D006224:Cricetinae; D004664:Encephalitis Virus, Japanese / Q000502:physiology*; D006801:Humans; D051379:Mice; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000502:physiology*; D014779:Virus Replication
D019253:Proto-Oncogene Proteins c-bcl-2