Propagation of hepatitis A virus in hybrid cell lines derived from marmoset liver and Vero cells.
To establish monkey liver cell lines with a high susceptibility to hepatitis A virus (HAV), marmoset (Saguinus labiatus) liver cells were fused with Vero cells deficient in hypoxanthine-guanine phosphoribosyltransferase and the resulting hybrid cells were selected in HAT medium. Of four hybrid cell lines obtained (S. 1a/Ve-1 to -4), three (S. 1a/Ve-1, -3 and -4) were equally susceptible to HAV infection. When inoculated with a virus isolated from marmoset liver tissue (10% liver tissue extract) or a faecal virus (10% stool extract) from a human hepatitis A patient, all susceptible cell lines showed a significant elevation of viral antigen activity as seen in radioimmunoassay and/or immunofluorescent antibody assays, at 4 to 6 weeks post-infection (p.i.) with the liver-derived inoculum and at 6 to 8 weeks p.i. with the stool-derived inoculum. In S. 1a/Ve-1 cells, a representative of the susceptible hybrid cell lines, full adaptation of HAV (liver tissue virus concentrate) to cell culture was attained after four serial passages. Thereafter, the virus grew to a plateau titre of 10(8.5) TCID50/ml at 7 days p.i. in a growth experiment. The infected cells showed no cytopathic effects but eventually a persistent infection was established when a saturated level of virus growth was reached.
Ashida
M
M
Department of Virology, Niigata University School of Medicine, Japan.
Hara
H
H
Kojima
H
H
Kamimura
T
T
Ichida
F
F
Hamada
C
C
D000818:Animals; D002143:Callitrichinae; D002459:Cell Fusion; D002460:Cell Line; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D006507:Hepatovirus / Q000254:growth & development*; D006822:Hybrid Cells / Q000382:microbiology*; D008099:Liver / Q000382:microbiology*; D014709:Vero Cells