Enhancement of hepatitis A propagation in tissue culture with 5,6‐dichloro‐1‐β‐D‐ribofuranosylbenzimidazole

Abstract

The adenosine analog 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) was found to increase the production of hepatitis A (HAV) antigen in two monkey kidney cell lines (Frhk-4 and Vero cells). DRB, a known inhibitor of the synthesis of messenger RNA, caused moderate changes in cell morphology. However, Frhk-4 cells could be maintained for several weeks at 80 microM of DRB, the concentration that caused maximal enhancement on HAV. DRB should be present from about the time of virus inoculation and its strongest effect was seen at low multiplicities of infection. Using radioimmunofocus assay it could be shown that DRB increased the amount of infectious virus. DRB treatment was applied in primary isolation of HAV from feces. In nine of ten strains HAV antigen expression was strongly increased and in six of the ten strains infectivity of harvested material increased by one 10log or more. DRB thus seems to be a useful enhancer of HAV growth in tissue culture.

Restricted replication of hepatitis A virus in cell culture: encapsidation of viral RNA depletes the pool of RNA available for replication

“…The plaque assay for HAV (1) was modified by the use of an overlay medium consisting of MEM containing 0.5% SeaPlaque agarose (FMC Corp., Marine Colloids Div., Rockland, Maine) and 25 ,uM 5,6-dichlorobenzimidazole riboside (Sigma Chemical Co., St. Louis, Mo. ), which has been reported to enhance HAV growth in cell culture (32). Addition of this drug results in a clear plaque morphology but, in our hands, had no significant effect on the yield of virus (D. A. Anderson, unpublished observations).…”

cDNA clone of hepatitis A virus encoding a virulent virus: induction of viral hepatitis by direct nucleic acid transfection of marmosets

“…We attempted to recover infectious HAV by inoculating FRhK-4 cell cultures with aqueous suspensions of feces that were positive by slot-blot hybridization analysis. To further increase the possibility of isolating HAV, we treated the cell cultures with 5,6-dichloro-1-3-D-ribofuranosylbenzimidazole, an adenosine analog which enhances the growth of wild-type HAV in cell culture (8,21). Cell cultures inoculated with the fecal suspensions became infected with HAV since the cells were subsequently stained with antibodies to HAV in an immunofluorescence microscopy assay (Table 2).…”

Ανοσογονικοτητα Και Ασφαλεια Εμβολιου Απο Αδρανοποιημενο Ιο Ηπατιτιδας Α

“…Μπορεί να αναπτυχθεί στους 32°C, στους 35°C ή και στους 37°C <15), Η ανάπτυξη του δεν αναστέλλεται με την επίδραση αντίίκών ουσιών (87,88,89). Μάλιστα βρέθηκε ότι μια γνωστή αντιϊκή ουσία η 5,6 Dichloro-1-ß-D-Ribofuranosylbenzimidazole προάγει αντί να αναστέλλει τον πολλαπλασιασμό του (90).…”