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Related Experiment Videos

Activation-dependent proteolytic degradation of polymorphonuclear CD11b.

P C Davey1, M Zuzel, A S Kamiguti

  • 1Department of Haematology, University of Liverpool, Liverpool, UK.

British Journal of Haematology
|December 21, 2000
PubMed
Summary

Strong stimulation causes polymorphonuclear (PMN) leucocytes to lose CD11b surface expression due to proteolytic degradation. This alteration affects PMN adhesion and may have significant pathophysiological importance.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • CD11b/CD18 is a key integrin on polymorphonuclear (PMN) leucocytes, crucial for their adhesion, migration, and phagocytosis.
  • This receptor is stored intracellularly and translocated to the cell surface upon stimulation.

Purpose of the Study:

  • To investigate the dynamic changes in CD11b surface expression on PMNs following strong stimulation.
  • To elucidate the mechanism behind the observed loss of CD11b surface expression and its functional consequences.

Main Methods:

  • Fluorescence-activated cell sorting (FACS) to detect CD11b surface expression using monoclonal antibodies.
  • Stimulation of PMNs with phorbol 12-myristate 13-acetate (PMA), N-formylmethionyl-leucyl-phenylalanine (FMLP), granulocyte macrophage-colony stimulating factor (GM-CSF), and platelet factor (PF) 4.

Related Experiment Videos

  • Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis for molecular characterization.
  • Proteinase inhibitor assays to identify the degrading enzyme.

    • Main Results:

    • Strong PMN stimulation led to initial CD11b upregulation followed by a time-dependent loss of the N-terminal epitope.
    • This loss occurred with potent stimuli (PMA, FMLP/GM-CSF/PF4) but not weak stimulation (FMLP alone).
    • CD11b degradation by a cell-associated serine proteinase was identified as the cause of epitope loss, paralleling changes in PMN adhesion to fibrinogen and ICAM-1.

    Conclusions:

    • Strong stimulation induces proteolytic degradation of the CD11b molecule in PMNs.
    • This alteration of CD11b affects PMN adhesion, suggesting significant pathophysiological implications in inflammatory conditions.