CD103 (aE integrin) is highly expressed on regulatory T cells (Tregs) found within skin. CD103 binds to E-cadherin, a molecule predominantly expressed by keratinocytes in the epidermis and around hair follicles. CD103 is thought to contribute to Treg retention within the skin. However, as the majority of Tregs are located within the dermis, the mechanism of this action is unclear. Moreover, while inflammation significantly increases the migration of intradermal Tregs, the contribution of CD103 to Treg localisation and motility is unclear. To examine this, we performed multiphoton microscopy of the skin of Foxp3-GFP mice treated with CD103 blocking antibodies to examine the contribution of CD103 to dermal Treg behaviour in uninflamed skin as well as during the peak (24 h) and resolution phases (48 h) of oxazolone-induced contact sensitivity (CS). Inhibition of CD103 did not alter Treg migration in untreated skin or at the peak of inflammation. In contrast, at 48 hours, when inflammation is resolving, inhibition of CD103 increased Treg migration. This result is consistent with CD103 acting to promote Treg retention in the skin at later phases of the inflammatory response. This coincided with the upregulation of E-cadherin on dermal dendritic cells and macrophages during the CS response. Using CD11c-YFP x Foxp3-GFP dual dendritic cell/Treg reporter mice, we demonstrated that at the 48 h CS timepoint, inhibition of CD103 reduced the percentage of Tregs interacting with dendritic cells, as well as the duration of the interactions between these cell types. Together these studies demonstrate that the role of CD103 in restraining Treg motility in inflamed skin varies according to the phase of the response, and that increased E-cadherin expression by leukocytes within the dermis may contribute to Treg retention and prolonged interactions with CD11c+ cells during the inflammatory response.