Dendritic cells (DCs) are a heterogeneous population of APCs with critical roles in T cell activation and immune regulation. We report in this study the identification and characterization of a novel subset of DCs resident in skin-draining peripheral lymph nodes of normal mice. This subset of CD11c highCD40highCD8αintermediate (int) DCs expresses the collagen-binding integrin, α1β1 and the E-cadherin-binding integrin, αEβ7. Although α1β1 and αEβ 7 are also expressed on CD11chighCD40 intCD8αhigh lymphoid DCs, CD11c highCD40highCD8αint DCs demonstrate preferential integrin-mediated adhesion to collagen and fibronectin. This DC subset most likely acquires expression of these integrins in peripheral lymph node, as this subset is not found in the spleen or mesenteric lymph node, and recent DC migrants from the skin lack expression of α1β 1 and αEβ7 integrins. Resident CD40high DCs express α1β1 integrin and colocalize with collagen in lymph nodes. When compared with CD11c highCD40highCD8αint DCs lacking expression of these integrins, the α1β1 +αEβ7+ DC subset exhibits more efficient formation of Ag-independent conjugates with T cells, and a decreased ability to acquire soluble Ag. Thus, the α 1β1 and αEβ7 integrins define a unique population of peripheral lymph node-derived DCs with altered functional properties and adhesive potential that localizes these cells to sites in lymph nodes where Ag presentation to T cells occurs.