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CX₃CR1⁺ and CD103⁺ dendritic cells (DCs) in intestinal lamina propria play a key role in mucosal immunity. However, the origin and the developmental pathways that regulate their differentiation in the lamina propria remain unclear. We showed that monocytes gave rise exclusively to CD103⁻CX₃CR1⁺ lamina propria DCs under the control of macrophage-colony-stimulating factor receptor (M-CSFR) and Fms-like thyrosine kinase 3 (Flt3) ligands. In contrast, common DC progenitors (CDP) and pre-DCs, which give rise to lymphoid organ DCs but not to monocytes, differentiated exclusively into CD103⁺CX₃CR1⁻ lamina propria DCs under the control of Flt3 and granulocyte-macrophage-colony-stimulating factor receptor (GM-CSFR) ligands. CD103⁺CX₃CR1⁻ DCs but not CD103⁻CX₃CR1⁺ DCs in the lamina propria constitutively expressed CCR7 and were the first DCs to transport pathogenic Salmonella from …