Using transmission electron microscopy, we demonstrate in this study the existence of intracellular Moloney murine leukemia virus particles in cytoplasmic vacuoles of chronically infected NIH/3T3 cells. Interferon (IFN) treatment for 48 h resulted in a 3-fold higher accumulation of these vacuolar virions. Intracellular virions and their accumulation in IFN-treated cells could also be shown by radioactive labeling of the cells for 48 h and isolation of labeled virus particles from the cytoplasmic fraction of the cells by banding in sucrose gradient. These virions contained typical viral RNA, proteins and glycoproteins, as determined by gel-electrophoresis. Despite their higher accumulation, the formation rate of the intracellular virions, as measured by their pulse labeling for 1 h with S35 -methionine, was found to be remarkably lower in IFN-treated cells. In order to elucidate this discrepancy, we followed the kinetics of the intracellular and extracellular virus appearance. We found that the inhibitory effect of IFN on the release of virus particles to the culture medium was much stronger than its effect on the formation of the intracellular virions. This finding can explain the accumulation of intracellular virus particles in IFN-treated cells despite their reduced formation rate.
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