Susceptibility of mouse primary cortical neuronal cells to coxsackievirus B.
Coxsackievirus B (CVB) is often associated with aseptic meningitis and encephalitis, but the six serotypes of CVB vary in their relative disease severity. To elucidate the detailed mechanisms of CVB-induced cytopathological effects, the morphological and biochemical characteristics caused by the CVB serotypes in mouse primary cortical neuronal cells were investigated. By 24 h post-infection, all CVB serotypes except CVB2 induced severe cytotoxic alterations, including a loss of neurites. Both fluorescence and transmission electron microscopy revealed CVB-induced morphological changes indicative of apoptosis, including heavily condensed nuclei, subsequent chromatin condensation into the periphery of the nuclei and oligonucleosomal DNA fragmentation. It was also found that infection with all six CVB serotypes led to productive virus replication, which was completed prior to an apoptotic signal. The caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone significantly inhibited nuclear changes associated with virus-induced apoptosis, but had less effect on virus-associated cytopathic effects and no effect on virus production. In contrast, the transcription inhibitor actinomycin D profoundly inhibited all three virus-induced events. Taken together, these findings demonstrate that all six CVB serotypes can efficiently replicate in mouse cortical neuronal cells and that productive replication of these CVBs, except for CVB2, induces multiple cytopathological effects, including apoptotic alterations.
Ahn
Jeonghyun
J
Departments of Microbiology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Choi
Jene
J
Departments of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Joo
Chul Hyun
CH
Departments of Microbiology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Seo
Ilseon
I
Departments of Microbiology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Kim
DongHou
D
Departments of Anatomy and Cell Biology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Yoon
Seung Yong
SY
Departments of Anatomy and Cell Biology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Kim
Yoo Kyum
YK
Departments of Microbiology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
Lee
Heuiran
H
Departments of Microbiology, University of Ulsan College of Medicine, Asan Medical Center, Songpa, PO Box 145, Seoul, Korea.
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D002540:Cerebral Cortex / Q000821:virology*; D002522:Chlorocebus aethiops; D003588:Cytopathogenic Effect, Viral; D003609:Dactinomycin / Q000494:pharmacology; D029822:Enterovirus B, Human / Q000472:pathogenicity*; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000821:virology*; D014709:Vero Cells; D014779:Virus Replication
D000590:Amino Acid Chloromethyl Ketones; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D003609:Dactinomycin