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Studies on the Mechanism of the Interferon-mediated Antiviral State to Vesicular Stomatitis Virus in Resting Mouse Peritoneal Macrophages

¹ Department of Experimental Medicine and Biochemical Sciences, II University of Rome ‘Tor Vergata’
and² Laboratory of Virology, Istituto Superiore di Sanità, Viale Regina Elena 299, Rome, Italy

We have analysed the expression of vesicular stomatitis virus (VSV) proteins in virus-infected freshly explanted mouse peritoneal macrophages (resistant to virus replication), macrophages aged in vitro (permissive for virus replication) and freshly explanted macrophages from mice treated with antibody to interferon (IFN) / (permissive for VSV replication). Our data showed that some VSV proteins (i.e. N/NS and G) were synthesized in virus-infected (1 p.f.u./cell) freshly harvested macrophages at early times after infection (3 to 6 h); the expression of such viral proteins was subsequently inhibited at 18 h post-infection. In contrast, a progressive increase in the expression of VSV proteins was observed in the macrophages aged in vitro and infected with VSV at 1 p.f.u./cell. Infection with a higher m.o.i. (16 p.f.u./cell) resulted in similar viral protein electrophoresis patterns for both aged macrophages and freshly explanted macrophages. Even at low m.o.i. a marked and progressive expression of all VSV proteins was observed in freshly harvested macrophages from mice treated with antibody to mouse IFN-/. Higher levels of oligo-2',5'-adenylate synthetase (2-5AS) were found in freshly harvested macrophages than in either aged macrophages or those from mice treated with antibody to IFN. No dsRNA-dependent 67K protein kinase was detected in freshly harvested macrophages or peritoneal cells from untreated mice or mice treated with poly(rI).poly(rC) or Newcastle disease virus. The following conclusions can be drawn from these results. Low levels of spontaneous IFN-/ are responsible for the time-dependent inhibition of VSV protein synthesis in virus-infected freshly harvested macrophages; high levels of 2-5AS (in the absence of detectable levels of 67K protein kinase) appear to correlate with the progressive inhibition of VSV proteins; this natural antiviral state is highly effective only at low m.o.i.

Keywords: interferon (MuIFN-/), NDV, macrophages

Received 16 November 1988; accepted 3 February 1989.