Ultrastructural localization of NADPH-oxidase activity in murine peritoneal macrophages during phagocytosis of Brucella. Correlation with the production of superoxide anions

The localization of the enzyme NADPH oxidase in mouse peritoneal macrophages unstimulated or after phagocytosis of Brucella suis was investigated by electron microscopy in normal mice and mice immunized against B. suis. The enzyme was clearly visualized on mitochondrial cristae, smooth endoplasmic reticulum, and the plasma membrane; its activity correlated mainly with the state of the endoplasmic reticulum which itself reflected macrophage activation. The enzyme turnover appeared to be accelerated after phagocytosis; the phagosome membrane was seldom stained by the enzyme reaction. These macrophages were also examined for the production of superoxide anions in vitro, either unstimulated or after phagocytosis. Phagocytosis increased the production of superoxide anions, especially in immunized animals. These results are discussed with regard to the role that the products of oxidative metabolism play in the inactivation of bacteria by phagocytic cells.