As major antigen-presenting cell, dendritic cells are essential in initiating T cell responses. They are found in secondary lymphoid organs and the periphery and can be divided into many different subsets that exert different functions. In addition to blood-derived DC subsets, lymph nodes can also contain a group of DC that migrate from the periphery to the lymph nodes. For skin-draining lymph nodes, the migratory populations consist of epidermis-derived Langerhans cells and dermal DC as well as CD103+ DC. In this study, we examined which DC subsets are involved in priming of Herpes simplex virus type 1 (HSV-1)-specific CD8+ T cells after flank infection. Using IRF8-/- mice as well as a mouse model of conditional depletion of Langerin+ cells we could show that CD8α+ DC and/or CD103+ DC are the main antigen-presenting subsets that prime HSV-1 specific CD8+ T cells in vivo. Next, we made use of the Rosa-DTA (R-DTA) mouse strain that carries a floxed-STOP cassette in front of a Diphtheria toxin subunit within the ROSA26 locus. By infecting R-DTA mice with HSV-1 expressing the Cre recominase the STOP cassette is selectively excised in HSV-1-infected cells resulting in Diphtheria toxin expression and cell death. This approach allowed us to selectively delete HSV-1-infected cells and eliminate potential antigen-presentation by infected DC. Using this R-DTA system we could show in vitro and in vivo that CD8+ T cell priming and subsequent expansion and acquisition of effector functions mainly occurred via cross-presentation. These findings are consistent with a model in which skin-resident DC that are either directly infected or have taken up virally-infected material migrate to the draining lymph nodes and hand their cargo over to lymph node-resident CD8α+ DC for priming of CD8+ T cells.