The innate immune response plays an essential role in the prevention of early viral dissemination. We used the lymphocytic choriomeningitis virus model system to analyze the role of tissue macrophages/Kupffer cells in this process. Our findings demonstrated that Kupffer cells are essential for the efficient capture of infectious virus and for preventing viral replication. The latter process involved activation of Kupffer cells by interferon (IFN)‐I and prevented viral spread to neighboring hepatocytes. In the absence of Kupffer cells, hepatocytes were not able to suppress virus replication, even in the presence of IFN‐I, leading to prolonged viral replication and severe T cell‐dependent immunopathology. Conclusion: Tissue‐resident macrophages play a crucial role in early viral capture and represent the major liver cell type exhibiting responsiveness to IFN‐I and providing control of viral replication. (HEPATOLOGY 2010)