Previous animal model experiments have shown a correlation between interferon gamma (IFN-␥) expression and both survival from infection with attenuated rabies virus (RABV) and reduction of neurological sequelae. Therefore, we hypothesized that rapid production of murine IFN-␥ by the rabies virus itself would induce a more robust antiviral response than would occur naturally in mice. To test this hypothesis, we used reverse engineering to clone the mouse IFN-␥ gene into a pathogenic rabies virus backbone, SPBN, to produce the recombinant rabies virus designated SPBN␥. Morbidity and mortality were monitored in mice infected intranasally with SPBN␥ or SPBN(؊) control virus to determine the degree of attenuation caused by the expression of IFN-␥. Incorporation of IFN-␥ into the rabies virus genome highly attenuated the virus. SPBN␥ has a 50% lethal dose (LD 50) more than 100-fold greater than SPBN(؊). In vitro and in vivo mouse experiments show that SPBN␥ infection enhances the production of type I interferons. Furthermore, knockout mice lacking the ability to signal through the type I interferon receptor (IFNAR ؊/؊ ) cannot control the SPBN␥ infection and rapidly die. These data suggest that IFN-␥ production has antiviral effects in rabies, largely due to the induction of type I interferons.
IMPORTANCE
Survival from rabies is dependent upon the early control of virus replication and spread. Once the virus reaches the central nervous system (CNS), this becomes highly problematic. Studies of CNS immunity to RABV have shown that control of replication begins at the onset of T cell entry and IFN-␥ production in the CNS prior to the appearance of virus-neutralizing antibodies.Moreover, antibody-deficient mice are able to control but not clear attenuated RABV from the CNS. We find here that IFN-␥ triggers the early production of type I interferons with the expected antiviral effects. We also show that engineering a lethal rabies virus to express IFN-␥ directly in the infected tissue reduces rabies virus replication and spread, limiting its pathogenicity in normal and immunocompromised mice. Therefore, vector delivery of IFN-␥ to the brain may have the potential to treat individuals who would otherwise succumb to infection with rabies virus. R abies virus (RABV) is the type species of the Lyssavirus genus in the Rhabdoviridae family. Its small, negative-stranded RNA genome contains only five true genes (1, 2). Although relatively simple, this zoonotic virus has a devastating impact worldwide. The majority of human rabies deaths occur in children in the developing world, and it is estimated that at least 55,000 humans die of rabies each year in Africa and Asia alone (3).Although RABV infection historically has been viewed as a death sentence once the virus reaches the brain, there is a small but growing number of humans who have survived rabies even though the virus entered the brain (4, 5). Due to such cases and to research using animal models of RABV infection (6-8), many believe that the immune system may be cap...