The majority of the response of cytotoxic T-lymphocytes (CTL) to lymphocytic choriomeningitis virus (LCMV) in H-2d mice is directed toward one epitope located on the nucleoprotein (NP, aa 118-126), and usually no primary responses to other epitopes are detectable. Previous studies have shown that thymic expression of lymphocytic choriomeningitis virus-nucleoprotein (LCMV-NP) in H-2d transgenic mice (Thy-NP mice) leads to deletion of high-affinity anti-LCMV-NP CTL by negative selection. Selection is incomplete, so that low-affinity NP-specific CTL pass through the thymus and are detectable in the periphery. To analyze the importance of interferon-gamma (IFN-gamma) in the ability of low-affinity antiviral CTL to clear an acute viral infection, double transgenic mice were generated that are IFN-gamma deficient and express the NP of LCMV in the thymus (Thy-NP x IFN-gamma -/- mice). When infected with LCMV, these bigenic mice were unable to clear the infection despite generating low-affinity primary antiviral CTL, and they became persistently infected. In contrast, IFN-gamma competent Thy-NP mice cleared LCMV within 7-8 days and IFN-gamma deficient mice that did not express NP in their thymus generated high-affinity CTL that terminated an acute LCMV infection within 10-12 days post-viral challenge. Persistently infected IFN-gamma deficient mice selectively depleted LCMV-specific CTL and displayed reduced levels of antigen-presenting cells in the spleen, and 60% of these mice died at 2-3 months postinfection. Thus, IFN-gamma is required for clearing an acute viral infection in the absence of a high-affinity CTL response. In the absence of IFN-gamma persistent viral infection results despite the presence of low-affinity CTL.