A Kinetic Model of Superoxide Production from Single Pulmonary Alveolar Macrophages

"DIGREGORIO, K. A., E. V. CILENTO, AND R. C. LANTZ. A kinetic model of superoxide production from single pulmonary alueolar macrophages. Am. J. Physiol. 256 (Cell Physiol. 25): C405-C412, 1989.-A kinetic model was developed to describe the production of superoxide (02) by single pulmonary alveolar macrophages (PAM). Model predictions were compared with experimental results obtained from single rat PAM. The 02 was quantified by measuring the reduction of nitro blue tetrazolium (NBT) to a diformazan precipitate (NBTH,) from video-recorded images of individual cells. The kinetic model considered three reactions: 1 ) the production of extracellular 02 from the reduction of oxygen by NADPH oxidase using intracellular NADPH as the substrate, 2) the subsequent dismutation of 02 to form H,O,, and 3) the reaction of 02 and NBT to form diformazan. NBT specificity for 02 was analyzed by comparing results in the presence and absence of superoxide dismutase (SOD) that catalyzes the dismutation of 02 to H,O,. Measured PAM heterogeneity was accounted for by varying the concentration of intracellular NADPH, its rate of depletion, and the concentration of intracellular NADPH oxidase in the kinetic model. Model predictions compared favorably with experimental results except when SOD was present. This discrepancy may be due to diffusional limitations because NBT is a relatively small molecule (818 mo! wt) compared with SOD (34,000 mol wt). In addition, the cell surface is both ruffled and negatively charged, which may introduce steric hindrances and/ or electrostatic effects, since SOD is also negatively charged, whereas NBT is positively charged.PULMONARY ALVEOLAR MACROPHAGES (PAM) protect the lungs by phagocytizing foreign debris and bacteria. This process is accompanied by the production of superoxide anion (02) at the surface of the plasma membrane (3, 31). Superoxide readily dismutates to form H20 2 and subsequently, other oxygen metabolites which have been shown to be both antibacterial and cytotoxic (21, 25, 35).Superoxide is produced from the reduction of extracellular O, at the surface of the cell membrane by NADPH oxidase by the following reaction (3, 4)"