The production of platelet-activating factor (PAF) by rat peritoneal cells was studied using as stimuli either monoclonal IgE, IgG1 or IgG2b anti-DNP (2,4-dinitrophenyl), and DNP-BSA. Peritoneal cells sensitized in vitro with any of these antibodies at concentrations higher than 10 nM and challenged with 1 microM DNP-BSA produced PAF. PAF production was also elicited by preformed IgE/ and IgG2b/DNP-BSA immune complexes, preferentially at a large antigen/antibody ratio. The production of PAF was unrelated to the activation of mast cells, since it occurred in populations depleted of mast cells by adherence to plastic dishes. Moreover, the release of [3H]serotonin from IgE-sensitized mast cells showed a time-course more rapid than PAF production and occurred in cells sensitized with IgE at concentrations lower than those required for PAF formation. In contrast, peritoneal cells sensitized with IgG1 and IgG2b failed to release [3H]serotonin. Rat peritoneal cells showed a significant ability to catabolize PAF by intracellular PAF-acetylhydrolase in view of both the amounts of enzyme activity assayed in cellular homogenates, and the 15-fold increase on controls of PAF quantities detected in peritoneal cells treated with phenylmethylsulfonyl fluoride (PMSF), a known inhibitor of PAF-acetylhydrolase. The PAF activity produced upon PMSF addition showed a retention time on reverse-phase HPLC which suggests structural identity to PAF produced by either immunological challenge or ionophore A23187. These data suggest that PAF formed during rat passive anaphylaxis reactions depends on the activation of mononuclear phagocytes. This production may be triggered by two types of low affinity receptors: Fc epsilon RII/CD23 and Fc gamma R. The ability of peritoneal cells to catabolize PAF by intracellular acetylhydrolase seems unaffected by immunological stimulation.