The classical model of bacterial killing by phagocytic cells has been recently challenged by questioning the toxic effect of oxygen products and attributing the fundamental role to K+ ions in releasing antimicrobial proteins within the phagosome. In the present study we followed O2.- production, changes of membrane potential, K + efflux, and bacterial killing in the presence of increasing concentrations of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium. Efficiency of bacterial killing was assessed on the basis of bacterial survival measured by a new semiautomated method. Very low rates of O2.- production were accompanied by significant membrane depolarization and K+ release and parallel improvement of bacterial killing. When O2.- production exceeded 20% of its maximal capacity, no further change was detected in the membrane potential and only minimal further K+ efflux occurred, yet bacterial survival decreased parallel to the increase of O2 .- production. The presented results indicate that both electrophysiological changes (depolarization and consequent ion movements) and the chemical effect of reactive oxygen species play a significant role in the killing of certain pathogens. The observation that an increase of membrane depolarization can compensate for decreased O2.- production may be important for potential therapeutic applications.
ASJC Scopus subject areas
- Cell Biology