ABSTRACT Human norovirus (HuNoV) causes acute gastroenteritis in immunocompetent hosts and chronic infection in immunocompromised individuals. Many recent studies of replication and innate immune responses following HuNoV infection have utilized epithelium-only human intestinal enteroids (HIEs), which lack immune cells. Here, we utilized an ex vivo enteroid-macrophage coculture model consisting of HIEs and different subtypes of human peripheral blood mononuclear cell-derived macrophages to better recapitulate in vivo gut biology and explore the role of macrophages in HuNoV replication and pathogenesis. We show that HuNoV infection in HIEs polarized on Transwells leads to bilateral release of the viral genome, with predominant apical virus release. Coculture with naïve M0, pro-inflammatory M1, or anti-inflammatory M2 macrophages does not change levels of HuNoV replication. We found that macrophages respond to HuNoV infection by phagocytosis of virus-infected cells with pro-inflammatory M1 macrophages exhibiting the
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