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

Nature volume 416, pages 291–297 (2002)Cite this article

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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Author notes

  1. Emer P. Reeves, Hui Lu and Jürgen Roes: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Molecular Medicine, University College London, 5 University Street, London, WC1E 6JJ, UK

    Emer P. Reeves, Hui Lu, Carlo G. M. Messina & Anthony W. Segal

  2. Centre for Molecular Medicine, Department of Physiology, University College London, 5 University Street, London, WC1E 6JJ, UK

    Steve Bolsover

  3. Centre for Molecular Medicine, Department of Anatomy, University College London, 5 University Street, London, WC1E 6JJ, UK

    Giorgio Gabella

  4. Windeyer Institute of Medical Sciences, University College London, 5 University Street, London, WC1E 6JJ, UK

    Jürgen Roes

  5. Institut de Biologie Structurale, 41 rue Horowitz, Grenoble, 38027, France

    Hugues Lortat Jacobs

  6. Ultrafast Laser and Spectroscopy Laboratory, Materials Science Centre, University of Groningen, Nijenborgh 4, Groningen, 9747 AG, The Netherlands

    Eric O. Potma

  7. Rayne Institute, St Thomas' Hospital, Lambeth Palace Road, London, SE1 7EH, UK

    Alice Warley

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  1. Emer P. Reeves
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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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