Intestinal mucosa is always exposed to foreign substances such as food antigens and microorganisms, thereby being protected by the mucosal immune system. Further, the enteric nervous system, which contains intrinsic primary afferent neurons, interneurons and motor neurons, regulate the intestinal secretary, motor and digestive functions independent of the central nervous system. To maintain the gut homeostasis, the enteric nervous system and mucosal immune system are assumed to functionally coordinate with each other using common bioactive substances and their receptors, however underlying mechanisms of these crosstalk have not been precisely understood. Here, we investigated the interaction between dendritic cells (DCs), conductor cells in acquired immune responses, and enteric neurons using an in vitro co-culture system. First, we immunohistochemically observed that DCs (CD11c+ cells) were located in close proximity to enteric nerve fibers in the colonic mucosa of normal mice. In addition, we established the co-culture system of bone marrow-derived DCs (BMDCs) and isolated enteric neurons from the small intestine of normal mouse, and demonstrated that the activation of enteric neurons with veratridine increased cytosolic free calcium concentration in BMDCs, suggesting that bioactive substances such as cytokines and neurotransmitters released from the enteric neurons activated BMDCs. Furthermore, we found that IL-6 mRNA was highly expressed in the enteric neurons compared with spleen and brain. These results suggested that the enteric neurons constitutively communicate with dendritic cells in the intestinal mucosa, and that IL-6 plays an important role in this interaction to maintain immunological and physiological homeostasis in the gut.