Dendritic cells (DCs), which maintain tolerance and orchestrate T cell immune responses, comprise a heterogeneous group of cells. For example, in the steady state, murine spleen contains pre-DC-derived CD8+ and CD8− conventional DCs. During inflammation, monocytes become activated and acquire some DC-like features such as expression of CD11c and MHCII. Although each of these cell types can present antigen, the relative efficiency of processing and presentation after antigen capture by different routes has not yet been systematically compared. To this end we administered OVA to various conventional DCs and activated monocytes by receptor-mediated endocytosis, pinocytosis or phagocytosis and measured internalization and presentation to MHCI and MHCII restricted T cells. We find that CD8− DCs are more efficient than any other type of antigen presenting cell tested in terms of presenting antigen to MHCII restricted T cells, irrespective of the route of antigen capture. In contrast both subsets of splenic DCs are highly effective in cross-presenting antigens to CD8+ T cells. DCs and activated monocytes cross-presented antigens delivered by DEC205-mediated endocytosis and pinocytosis. However, DCs differ from activated monocytes in that the latter are several orders of magnitude less efficient in presenting antigens captured by phagocytosis to CD8+ or CD4+ T cells. We conclude that DCs derived from pre-DCs differ from monocyte-derived cells in that DCs process and present antigens efficiently irrespective of the route of antigen capture. Our observations have significant implications for understanding initiation of immune responses and vaccination strategies targeting DCs and activated monocytes.