Lymphocyte proliferation, mobility and longevity makes them prime targets for viral infection. Myeloid cells that process and present environmental antigens to lymphocytes are consequently an important line of defence. Subcapsular sinus macrophages (SSM) filter the afferent lymph and communicate with B cells. How they interact with B cell-tropic viruses is unknown. We analyzed their encounter with Murid Herpesvirus-4 (MuHV-4), an experimentally accessible gamma-herpesvirus related to the Kaposi's Sarcoma-associated Herpesvirus. MuHV-4 disseminates via lymph nodes, and intranasally or subcutaneously inoculated virions readily infected SSM. However this infection was poorly productive. SSM depletion with clodronate-loaded liposomes or with diphtheria toxin in CD169-diphtheria toxin receptor transgenic mice increased B cell infection and hastened viral spread to the spleen. Dendritic cells provided the main route to B cells, and SSM slowed host colonization apparently by absorbing virions non-productively from the afferent lymph.