To investigate the influences of acetaminophen (APP) on the immunotoxicity of ethanol (EtOH) in ICR mice, APP at a dose of 100 mg/kg was orally administered to mice daily for 28 consecutive days. Mice treated with EtOH were given freely with 20% w/v EtOH during the experimental period, and normal mice were given vehicle. The results of this study are summarized as follows: the combination of APP and EtOH significantly decreased the circulating leukocytes and the relative weights of liver, spleen and thymus, compared with the treatment of EtOH alone. In mice receiving the combination of AAP and EtOH when compared with the treatment of EtOH alone, there were also significant reductions in the splenic plaque forming cells (PFC) and hemagglutination (HA) titers to sheep red blood cells (SRBC), and the secondary IgG antibody response to bovine serum albumin (BSA). A tendency toward suppression of delayed-type hypersensitivity (DTH) reaction and phagocytic activity was also observed in the combination of AAP and EtOH. In addition, the combination of AAP and EtOH greatly increased serum alanine aminotransaminase (ALT) and total protein levels, compared with the treatment of EtOH alone. Significant decreases in serum albumin and A/G ratio were observed in EtOH alone-fed mice compared with those in normal animals, and their reductions were further induced in mice treated with AAP and EtOH. These findings indicate that EtOH-induced immunotoxicity is aggravated by the combination of APP and EtOH.