Chemokine-directed migration of leukocyte subsets may contribute to the qualitative differences between systemic and mucosal immunity. Here, we demonstrate that in mice lacking the chemokine receptor CCR6, dendritic cells expressing CD11c and CD11b are absent from the subepithelial dome of Peyer's patches. These mice also have an impaired humoral immune response to orally administered antigen and to the enteropathic virus rotavirus. In addition, CCR6^−/− mice have a 2-fold to 15-fold increase in cells of select T lymphocyte populations within the mucosa, including CD4⁺ and CD8⁺ αβ−TCR T cells. By contrast, systemic immune responses to subcutaneous antigens in CCR6^−/− mice are normal. These findings demonstrate that CCR6 is a mucosa-specific regulator of humoral immunity and lymphocyte homeostasis in the intestinal mucosa.