Experiments were performed to evaluate the in vitro effects of Escherichia coli lipopolysaccharide on viability and function of human alveolar macrophages. Alveolar macrophages were obtained by fiberoptic bronchoscopy and saline bronchial lavage from 12 normal, nonsmoking volunteers. Cells were incubated with different concentrations of E. coli endotoxin for 1 and 24 h. Endotoxin (10 microgram/ml and more) was cytotoxic for alveolar macrophages after 24 h of incubation and induced significant inhibition of phagocytosis, adherence, and spreading. The effects of endotoxin on alveolar macrophage viability and function were dose and time dependent and were not influenced by indomethacin. Thus, human alveolar macrophages, like other mononuclear phagocytes, are extremely sensitive to endotoxin effects; these observations may be relevant in conditions in which endotoxin may be in contact with alveolar macrophages in vivo: endobronchial infections with gram-negative organisms, byssinosis, chronic bronchitis of grain handles, and humidifier fever.