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Repeated 'superinduction' of interferon in human diploid fibroblast cultures
Abstract:
Human diploid fibroblast cultures induced to make interferon by the combination of polyriboinosinic acid-polyribocytidylic adic, cycloheximide and actinomycin D degenerate thereafter, owing to the irreversible nature of the inhibition induced by actinomycin D. However, cultures superinduced with the DNA-dependent RNA synthesis inhibitor 5,6-dichloro-I-beta-D-ribofuranosylbenzimadazole (DRB) survive, owing to the reversible nature of the inhibition induced by DRB, and can again be superinduced on several occasions.
Insights
Superinducing human cells with actinomycin D causes irreversible damage, leading to cell death. However, using 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) allows for reversible inhibition, enabling repeated interferon superinduction in cell cultures.
Area of Science:
- Molecular Biology
- Virology
- Cell Biology
Background:
- Interferon induction is crucial for antiviral defense.
- Previous methods using actinomycin D resulted in irreversible cell damage.
- The need for a method allowing repeated interferon induction without cell toxicity.
Purpose of the Study:
- To investigate the effect of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) on interferon superinduction in human diploid fibroblast cultures.
- To compare the reversibility of inhibition between DRB and actinomycin D in the context of interferon superinduction.
- To determine if DRB allows for repeated superinduction cycles.
Main Methods:
- Human diploid fibroblast cultures were used.
- Cells were induced to produce interferon using polyriboinosinic acid-polyribocytidylic acid.
- Inhibitors cycloheximide, actinomycin D, and 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) were applied.
- Cell survival and the ability for repeated superinduction were assessed.
Main Results:
- Superinduction with polyriboinosinic acid-polyribocytidylic acid, cycloheximide, and actinomycin D led to irreversible cell degeneration.
- Superinduction with polyriboinosinic acid-polyribocytidylic acid, cycloheximide, and DRB resulted in cell survival.
- DRB-treated cultures could be successfully superinduced on multiple occasions, demonstrating reversible inhibition.
- Actinomycin D-induced inhibition was irreversible, preventing subsequent superinduction.
Conclusions:
- The DNA-dependent RNA synthesis inhibitor DRB provides a reversible block, unlike the irreversible block by actinomycin D.
- DRB enables repeated interferon superinduction in human fibroblast cultures without causing cell death.
- This finding offers a more viable method for studying and utilizing interferon induction.