Mouse spleen contains three distinct mature dendritic cell (DC) populations (CD4+ 8−, CD4− 8−, and CD4− 8+) which retain a capacity to take up particulate and soluble Ags. Although the three splenic DC subtypes showed similar uptake of injected soluble OVA, they differed markedly in their capacity to present this Ag and activate proliferation in OVA-specific CD4 or CD8 T cells. For class II MHC-restricted presentation to CD4 T cells, the CD8− DC subtypes were more efficient, but for class I MHC-restricted presentation to CD8 T cells, the CD8+ DC subtype was far more effective. This differential persisted when the DC were activated with LPS. The CD8+ DC are therefore specialized for in vivo cross-presentation of exogenous soluble Ags into the class I MHC presentation pathway.