Mast cells are currently recognized as effector cells in many settings beyond just allergic reactions, including innate immunity, autoimmunity, chronic inflammatory disorders and atherosclerosis. Signaling pathways of the mast cell response have been widely explored in the past but these are still linked with single axes, such as the high affinity IgE receptor FcεRI, presumably an exclusive determinant of the magnitude of the response to allergen. By contrast, the T cell receptor is viewed as a rich complex of stimulatory and co-stimulatory molecules, setting an array of thresholds to ensure a highly regulated response. Recent observations show that mast cells express various classes of co-stimulatory molecules that modulate their function. These molecules might therefore contribute to the outcome of mast cell-associated pathologies, and constitute new therapeutic targets in such diseases.