Phosphate-dependent glutaminase (PDG) activity, a key enzyme of glutamine metabolism, was determined in neutrophils obtained from the intra-peritoneal cavity (PC) or bronchoalveolar space (BAS) after administration of 1 ml or 100 μl, respectively of saline, glycogen solution (1%) or lipopolysaccharide (LPS 0.1 mg (100 μl)−1). Neutrophils were obtained by lavage of both sites with 20 ml saline 24 h after the administration of the stimuli. Glycogen and LPS, depending on the site the cells were obtained from, differently modulated PDG activity. Cells from BAS stimulated by glycogen or LPS had raised PDG activity to 30.5 ± 5.2 and 42.7 ± 12.1 nmol min−1 mg−1 protein, respectively, when compared with saline (9.1 ± 0.9 nmol min−1 mg−1 protein); mean ± SEM. On the other hand, cells from PC showed different PDG activity: 52.0 ± 12.6 nmol min−1 mg−1 for saline, 36.5 ± 9.5 nmol min−1 mg−1 for glycogen, and 76.6 ± 11.2 nmol min−1 mg−1 for LPS; mean ± SEM. Therefore, PDG activity varies with the site from which neutrophils are obtained and the stimulus imposed. The effect of glutamine on nitric oxide (NO) and tumour necrosis factor (TNF) production by peritoneal neutrophils, obtained after glycogen administration, cultured in the presence of LPS (0.5 μg ml−1) was also examined. The addition of glutamine at concentrations varying from 2 to 20 mM did not markedly affect NO production. Glutamine alone at 2 mM did not modify the production of TNF but in the presence of LPS caused a significant decrease. So, glutamine may preserve the function of neutrophils during infections and injuries. Copyright © 2001 John Wiley & Sons, Ltd.