Fifty-four healthy humans, selected for their acute airway responsiveness to cotton dust, had spirometric tests immediately before and after 6 hours of exposure to card-generated cotton dust from seven different cottons (of several grades and growing regions). During exposures, we measured airborne concentrations of viable fungi and bacteria (total and gram negative), vertically elutriated gravimetric dust, and vertically elutriated endotoxin. Correlation between each of these five exposure indices and exposure-related acute changes in forced expiratory volume in 1 s showed a statistically significant relationship between all of the indices except concentration of viable fungi. Of the other four indices, endotoxin was the most highly correlated (r = -0.94; p less than 0.00001), and gravimetric dust was the least correlated (r = -0.34; p less than 0.05). These findings suggest that gram-negative endotoxin may play a major role in the acute pulmonary response to inhaled cotton dust.