Replication and plaque formation of swine hemagglutinating encephalomyelitis virus (67N) in swine cell line, SK-K culture.
Swine hemagglutinating encephalomyelitis virus (HEV), 67N strain, adapted to suckling mouse brain, grew readily in a porcine cell line, SK-K cell culture with cytopathic effect (CPE) consisting of syncytium formation and detachment of fused cells and round cells from glass surface. After further passages in SK-K cell monolayers with undiluted culture fluid, CPE developed earlier and became complete within 48 h postinoculation (p.i.). Viral specific antigen was detected in the cytoplasm of the infected SK-K cells by indirect immunofluorescence using rabbit antiserum against the mouse-passaged virus. The SK-K-passaged virus as well as the original mouse-passaged virus formed clear plaques on SK-K cell monolayers under simple overlay medium. The plaque assay system for HEV 67N was established by studying various factors influencing the plaque formation in the SK-K cell cultures. By this system more than 10(6) PFU/0.2 ml of the virus yield was detected in the fluid phase of the infected cultures at 48 h p.i. The SK-K-passaged virus caused fatal infection in 4-week-old mice by intracerebral inoculation, but was inhibited by rabbit antiserum against the mouse-passaged virus. Plaque formation and hemagglutinating activity of the virus were specifically inhibited by antisera against the mouse-passaged and SK-K-passaged 67N virus.
Department of Veterinary Microbiology, Iwate University, Morioka, Japan.
N
N
Hirano
K
K
Ono
H
H
Takasawa
T
T
Murakami
S
S
Haga
D000818:Animals; D002460:Cell Line; D003470:Culture Media; D003588:Cytopathogenic Effect, Viral; D004668:Encephalitis Viruses / Q000254:growth & development*; D004671:Encephalitis, Arbovirus / Q000473:pathology / Q000662:veterinary; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D006383:Hemadsorption Inhibition Tests / Q000662:veterinary; D006385:Hemagglutination Inhibition Tests / Q000662:veterinary; D008297:Male; D051379:Mice; D009500:Neutralization Tests / Q000662:veterinary; D013552:Swine / Q000382:microbiology*; D013553:Swine Diseases / Q000473:pathology; D010948:Viral Plaque Assay / Q000662:veterinary; D014779:Virus Replication
D003470:Culture Media