Rat peritoneal macrophages were incubated in the presence of beta-naphthylamine (beta-NA), a well known carcinogenic agent, and some parameters of respiratory burst were studied. beta-NA induced a time- and dose-dependent stimulation of superoxide anion (O-2) production, and this enhancement was suppressed by the addition of superoxide dismutase enzyme. Also, no cooperative effect between beta-NA and phorbol 12-myristate 13-acetate was observed. Other observations were as follows: (i) the simultaneous presence of polymyxin B, and staurosporine inhibitors of protein kinase C, inhibited beta-NA-dependent O-2 production; (ii) NADPH-oxidase contained in postnuclear fraction from beta-NA-incubated macrophages showed a greater activity than control fractions; (iii) the stimulation of O-2 production elicited by beta-NA was several-fold enhanced in activated macrophages compared to resident cells. These data suggest that beta-NA produces the activation of NADPH-oxidase through protein kinase C.