Antigen presenting cells (APC) of the intestinal tract are in continuous contact with a diverse array of microorganisms and food antigens, orchestrating a delicate balance to maintain immune tolerance to harmless antigens while initiating potent immunity to invading pathogens. While the mysteries of the former are quickly unravelling, it remains unclear how APC contribute to active immunity in the intestine. To gain a better understanding of the nature and function of APC in an environment conducive to an active immune response, we performed phenotypic and functional analysis of intestinal APC in steady state and after oral administration of the mucosal adjuvant cholera toxin (CT). In steady state, macrophage and dendritic cell (DC) populations are distinguished by the mutually exclusive expression of CD64 and CD24 respectively. Although the number and frequency of intestinal macrophage and DC populations did not change following CT treatment, existing CD24+ CD103+ DC upregulate cell surface expression of co-stimulatory molecules and the phosphatidylserine receptor T cell immunoglobulin mucin-4 (Tim-4). CD103+ Tim-4+ DC showed increased uptake of apoptotic material and soluble antigen in vitro compared CD103+ Tim-4- DC and could be isolated from the draining lymph node after CT treatment. Notably, upregulation of Tim-4 by DC in response to CT was independent of TLR4 signalling and gut microbiota. Taken together, our results suggest that activation with a mucosal adjuvant induced functional changes to the existing intestinal DC to promote the priming of mucosal immunity.