The ability of polymorphonuclear neutrophils (PMNs) and monocytes (Mφ) to produce reactive oxygen species (ROS) has been related closely to their potential in the killing of microorganisms. Ethanol has been shown to impair the generation of ROS in these phagocytes after stimulation with some immunogens and to increase the susceptibility of alcohol abusers to infectious diseases. As endotoxemia is common in alcohol abusers, we investigated the effect of ethanol (21.7 mmol/liter) on the luminol-amplified chemiluminescence of PMNs and Mφ after endotoxin stimulation and the release of tumor necrosis factor alpha (TNF-α) from Mφ. Further, the efficacy of ethanol to inactivate chemically generated ROS was tested. Significant stimulation of ROS release occurred at endotoxin concentrations of 1 ng/ml or higher in both PMNs and Mφ. Ethanol significantly suppressed the formation of ROS in both cell types, the decrease being more pronounced in Mφ (- 73.8%) than in PMNs (-45.7%). The correlation between endotoxin concentration and the amount of released ROS showed a dose-dependent, sigmoidal course. Concentrations of endotoxin necessary for half-maximum stimulation were nearly identical (6 to 8 ng/ml) in both PMNs and Mφ, independence of the presence of ethanol. In contrast to ROS formation, ethanol had no effect on the amount of TNF-α produced by endotoxin-stimulated Mφ. Ethanol was shown to unable to decrease the levels of chemically generated ROS under physiological conditions. Therefore, ethanol cannot be assumed to be an 'antioxidative' compound but rather seems to modify processes of endotoxin recognition, intracellular signal transduction, or metabolism.