ABSTRACT Powassan virus (POWV) is an emerging tick-borne flavivirus that causes neuroinvasive disease, including encephalitis, meningitis, and paralysis. Similar to other neuroinvasive flaviviruses, such as West Nile virus (WNV) and Japanese encephalitis virus (JEV), POWV disease presentation is heterogeneous, and the factors influencing disease outcome are not fully understood. We used Collaborative Cross (CC) mice to assess the impact of host genetic factors on POWV pathogenesis. We infected a panel of Oas1b -null CC lines with POWV and observed a range of susceptibility phenotypes, indicating that host factors other than the well-characterized flavivirus restriction factor Oas1b modulate POWV pathogenesis in CC mice. Among Oas1b -null CC lines, we identified multiple highly susceptible lines (0% survival), including CC071, and a single resistant line (78% survival), CC045. Susceptibility phenotypes generally were concordant among neuroinvasive flaviviruses, although we identified one line, CC006, that was resistant specifically to JEV, suggesting that both pan-flavivirus and virus-specific mechanisms contribute to susceptibility phenotypes in CC mice. We found that POWV replicated to higher titers in bone marrow-derived macrophages from CC071 mice compared to CC045 mice, suggesting that resistance could result from cell-intrinsic restriction of viral replication. Although serum viral loads at 2 days post-infection were equivalent between CC071 and CC045 mice, clearance of POWV from the serum was significantly slower in CC071 mice. Furthermore, CC045 mice had significantly lower viral loads in the brain at 7 days post-infection compared to CC071 mice, suggesting that reduced CNS infection contributes to the resistant phenotype of CC045 mice. IMPORTANCE Neuroinvasive flaviviruses, such as WNV, JEV, and POWV, are transmitted to humans by mosquitoes or ticks, can cause neurologic disease, such as encephalitis, meningitis, and paralysis, and can result in death or long-term sequelae. Although potentially severe, neuroinvasive disease is a rare outcome of flavivirus infection. The factors that determine whether someone develops severe disease after flavivirus infection are not fully understood, but host genetic differences in polymorphic antiviral response genes likely contribute to disease outcome. We evaluated a panel of genetically diverse mice and identified lines with distinct outcomes following infection with POWV. We found that resistance to POWV pathogenesis corresponded to reduced viral replication in macrophages, more rapid clearance of virus in peripheral tissues, and reduced viral infection in the brain. These susceptible and resistant mouse lines will provide a system for investigating the pathogenic mechanisms of POWV and identifying polymorphic host genes that contribute to resistance.

Neuroinvasive flavivirus pathogenesis is restricted by host genetic factors in Collaborative Cross mice, independently of Oas1b