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Killing activity of neutrophils is mediated through activation of proteases by K+ flux

Abstract

According to the hitherto accepted view, neutrophils kill ingested microorganisms by subjecting them to high concentrations of highly toxic reactive oxygen species (ROS) and bringing about myeloperoxidase-catalysed halogenation. We show here that this simple scheme, which for many years has served as a satisfactory working hypothesis, is inadequate. We find that mice made deficient in neutrophil-granule proteases but normal in respect of superoxide production and iodinating capacity, are unable to resist staphylococcal and candidal infections. We also show that activation provokes the influx of an enormous concentration of ROS into the endocytic vacuole. The resulting accumulation of anionic charge is compensated for by a surge of K+ ions that cross the membrane in a pH-dependent manner. The consequent rise in ionic strength engenders the release of cationic granule proteins, including elastase and cathepsin G, from the anionic sulphated proteoglycan matrix. We show that it is the proteases, thus activated, that are primarily responsible for the destruction of the bacteria.

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Figure 1: Susceptibility of protease-deficient mice to bacterial and fungal infection.
Figure 4: Vacuolar swelling and its control.
Figure 2: Appearance, concentration and buffering capacity of granule contents in the phagocytic vacuole.
Figure 3: K+ and 86Rb transport associated with the generation of O-2 in neutrophils.
Figure 5: Activation of the microbicidal proteases requires K+ and neutral pH.

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Acknowledgements

We thank W. Gratzer, P. Rich, F. Ashcroft, M. Brand, R. Vaughan-Jones and M. Duchen for discussions; E. Hawe for statistical analysis; A. Hankin and E. C. Davis for technical assistance; and A. Scott for illustration and art work. The Wellcome Trust and Chronic Granulomatous Disease Research Trust provided financial support.

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Correspondence to Anthony W. Segal.

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Reeves, E., Lu, H., Jacobs, H. et al. Killing activity of neutrophils is mediated through activation of proteases by K+ flux. Nature 416, 291–297 (2002). https://doi.org/10.1038/416291a

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