PROPAGATION OF HEPATITIS-A VIRUS IN A RENAL-CELL LINE JTC-12.P3 OF CYNOMOLGUS MONKEY ORIGIN

Human hepatitis A virus (HAV) derived from 10 % HAV infected marmoset liver homogenate and faeces from acute hepatitis A was successfully pr opagated in vitro in a new cell line, JTC-12.P3. The cell line origina ted from the renal cortex of cynomolgus monkey which was adapted to gr owth in a serum free, protein free, chemically defined synthetic mediu m. Replication of the virus was followed by solid phase RIA, immunoflu orescent staining, and immunoelectron microscopy. The propagation of H AV occurred over several passages, with the 1st and 2nd passages requi ring at least 8 weeks each. However, with the increasing serial passag e of virus, the period needed to detect it was shortened, suggesting t he adaptation of HAV to the cells. The identity of the newly synthetiz ed virus particles with HAV was established by immunoelectron microsco py and immunofluorescent blocking effect with human convalescent serum . The HAV propagated in JTC-12.P3 cells banded predominantly at a dens ity of 1.32 g/cm3 in CsCl gradient. The infected cells showed no speci fic signs of CPE. Ultrastructurally, clusters of virus particles 27 nm in diameter were observed mainly in the lysosomal vesicles and freely in crystalline array in the cytoplasm, too. Addition of 0.1 % of vari ous anti-HAV negative sera or of prostaglandin E1 to the culture mediu m caused accelerated propagation of HAV.