Protein analysis of defective interfering lymphocytic choriomeningitis virus and persistently infected cells.

Defective interfering (DI) lymphocytic choriomeningitis virus (LCMV) was purified from the culture fluids of BHK 21/13S and L-929 cells persistently infected with LCMV. DI LCMV sedimented in renografin-76 gradients to a density slightly less than standard (S) LCMV (1.13 vs 1.14 g/cm³). Polyacrylamide gel electrophoretic analysis of [³⁵S]methionine-labeled DI virus revealed a major 63,000-dalton polypeptide corresponding to the S virion nucleoprotein (NP), and two minor polypeptides corresponding to the S virion 54,000? and 35,000-dalton glycopeptides. No differences in polypeptide composition were detected between the DI and S virions. Exposure of cells to DI virus before S virus challenge inhibited the intracellular synthesis of the NP. Cells persistently infected with LCMV released no detectable S virus or temperature-sensitive mutants but did release DI LCMV. The production of DI LCMV by these cultures was 10? to 100-fold lower than S virus production during acute infections. These persistently infected cells contained intracytoplasmic NP, detectable by immunofluorescence, but its rate of synthesis was too low to be detected by the radiolabeling methods used. Although present in the cytoplasms, detectable viral antigens were absent from the cell membranes of many of these persistently infected cells. Thus, cells persistently infected with LCMV produce relatively low levels of DI virus which can inhibit viral protein synthesis. These factors may act to render infected cells resistant to immunosurveillance mechanisms during persistent infections in vivo.