Measurement of Superoxide Release from Single Pulmonary Alveolar Macrophages

"An electro-optical method was used to quantify superoxide (SO) release from single rat pulmonary alveolar macrophages (PAM) exposed in vitro to respirable quartz and kaolin dusts and in vivo to quartz. Release was determined by measuring the reduction of nitroblue tdrazolium (NBT) to a diformazan precipitate at 550 nm from video-recorded images of individual cells. In vitro exposure to 0.025 or 0.05 mg/ml kaolin for 40 minutes resulted in no significant differences in either the total diformazan produced (max) or the maximum rate of diformazan production (R)./n vitro exposure to 0.025 mg/ml c1uartz decreased max 38 percent while no significant change was found using 0.05 mg/ml. However, R was decreased 31 percent and 24 percent for the low and high dose, respectively. In contrast, PAM obtained from animals exposed in vivo to quartz dust for two or four weeks in inhalation chambers exhibited increased production of diformazan. However, only PAM analyzed three days post-exposure showed significant increases, while PAM from animals sacrificed ten or 31 days post-exposure were not significantly different from control. The combined in vitro and in vivo quartz results might suggest that there is an initital acute response to exposure that decreases the ability of PAM to produce SO, followed by recruitment of production of PAM with increased SO producing capability. It appears that removal of animals from the source of exposure permits PAM to return to control levels of SO production by31 days. Whether this would be true for chronic exposure remains to be elucidated.IntroductionPulmonary alveolar macrophages (PAM) protect the lungs by phagocytizing foreign debris and bacteria.(1) This process involves ingestion as well as destruction of foreign matter. Phagocytosis usually is accompanied by a respiratory burst which increases cellular oxygen consumption leading to the release of highly reactive oxygen metabolites.(2) These agents have been implicated in the killing of bacteria by phagocytes, but they have also been shown to be toxic and,have been linked to cancer, emphysema, arthritis, diabetes, and other diseases.(3) Toxic effects may be a result of 1) an abnormally low production of metabolites, resulting in damage to lung tissue by the respired bacteria and/or dusts; or 2) an abnormally high production resulting in direct damage to lung tissue by metabolites themselves."