Abstract
Neutrophil leukocytes have a pivotal function in innate immunity. Dogma dictates that the lethal blow is delivered to microbes by reactive oxygen species (ROS) and halogens, products of the NADPH oxidase, whose impairment causes immunodeficiency. However, recent evidence indicates that the microbes might be killed by proteases, activated by the oxidase through the generation of a hypertonic, K+-rich and alkaline environment in the phagocytic vacuole. Here we show that K+ crosses the membrane through large-conductance Ca2+-activated K+ (BK(Ca)) channels. Specific inhibitors of these channels, iberiotoxin and paxilline, blocked oxidase-induced 86Rb+ fluxes and alkalinization of the phagocytic vacuole, whereas NS1619, a BK(Ca) channel opener, enhanced both. Characteristic outwardly rectifying K+ currents, reversibly inhibited by iberiotoxin, were demonstrated in neutrophils and eosinophils and the expression of the alpha-subunit of the BK channel was confirmed by western blotting. The channels were opened by the combination of membrane depolarization and elevated Ca2+ concentration, both consequences of oxidase activity. Remarkably, microbial killing and digestion were abolished when the BK(Ca) channel was blocked, revealing an essential and unexpected function for this K+ channel in the microbicidal process.
MeSH Terms
Calcium/metabolism
Electric Conductivity
Eosinophils/cytology,drug effects,immunology,metabolism
Humans
Hydrogen-Ion Concentration
Immunity, Innate/drug effects
Ion Transport/drug effects
Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
Large-Conductance Calcium-Activated Potassium Channels
Membrane Potentials/drug effects
Neutrophils/cytology,drug effects,immunology,metabolism
Oxygen Consumption/drug effects
Phagocytosis/drug effects
Potassium/metabolism
Potassium Channels, Calcium-Activated/antagonists & inhibitors,metabolism
Rubidium Radioisotopes
Staphylococcus aureus/immunology
Tetradecanoylphorbol Acetate/pharmacology
Vacuoles/drug effects,metabolism
Chemicals
Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
Large-Conductance Calcium-Activated Potassium Channels
Potassium Channels, Calcium-Activated
Rubidium Radioisotopes
Tetradecanoylphorbol Acetate
Potassium
Calcium
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Ahluwalia Jatinder
Department of Medicine University College London, Gower Street, London WC1E 6BT, UK.
Tinker Andrew
Clapp Lucie H
Duchen Michael R
Abramov Andrey Y
Pope Simon
Nobles Muriel
Segal Anthony W
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