In addition to having anti-sympathotonic effects, beta-blockers are thought to have some adrenoceptor-independent properties. Such ancillary effects are described for carvedilol acting as oxygen radical scavenger and for propranolol which blocks protein kinase C and phosphatidate phosphohydrolase. The goal of our in vitro experiments was to identify ancillary effects of the widely used beta-blockers metoprolol and atenolol in neutrophils. Neutrophil chemotaxis was tested using the leading front assay in a modified Boyden microchemotaxis chamber. Respiratory burst activity was detected fluorometrically. Inhibition of protein kinase C activity was tested with purified alpha-, beta- and gamma-isoenzyme preparation. Metoprolol dose-dependently inhibited formyl peptide-stimulated neutrophil chemotaxis and formylpeptide- and phorbol myristate acetate-triggered oxygen free radical production. These actions were not affected by the competitive presence of the beta-receptor agonist, orciprenaline. Effects of metoprolol, as well as of propranolol, and the signaling enzyme blockers were strongly time dependent. Propranolol mimicked effects of staurosporine on respiratory burst, whereas the effects of metoprolol were similar to bisindolylmaleimide, a specific protein kinase C blocker. Atenolol, a hydrophilic beta-blocker, neither affected neutrophil chemotaxis nor respiratory burst. In a cell-free system, metoprolol did not interfere with the activity of the purified protein kinase C alpha-, beta- and gamma-isoenzymes. Adrenoceptor-independent inhibition of neutrophil chemotaxis and free radical production is a novel mode of action of metoprolol that may be relevant for beneficial effects ot the beta-blocker in heart failure and endothelial preconditioning.