Stimulated mast cells produce and release adenosine, and the release of mast cell mediators is potentiated by adenosine, yet very little is known regarding mast cell purine metabolism. Because 5-amino-4-imidazolecarboxamide riboside (AICA riboside) has been shown to alter adenosine metabolism and accelerate the repletion of ATP pools in other tissues, its effects on mast cell function were examined. Neither simultaneous addition of A23187 and AICA riboside nor a 1-hr preincubation with AICA riboside altered mast cell beta-hexosaminidase release to an appreciable degree. However, mouse bone marrow-derived mast cells cultured for 2 or more days in the presence of 1-100 microM AICA riboside exhibited a markedly attenuated mediator release response to A23187 compared to control cells with or without the additional presence of adenosine. IgE-mediated leukotriene C4 generation from AICA riboside-exposed mast cells was even more profoundly inhibited without affecting cell viability or resting mediator content. An unusual ribonucleotide triphosphate previously identified in folate-depleted cells, 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-triphosphate (ZTP), has been identified in AICA riboside-treated mast cells as well. Although the mechanism of this global inhibition of mast cell mediator release by chronic AICA riboside treatment is not clear, alterations in mast cell purine metabolism may prove to be important in the treatment of allergic diseases.