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Updated: May 9, 2026

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions
11:22

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

Published on: August 23, 2013

Degranulating stimuli decrease the neagative surface charge and increase the adhesiveness of human neutrophils

J I Gallin

    The Journal of Clinical Investigation
    |February 1, 1980
    PubMed

    Insights

    Neutrophil degranulation, triggered by various agents, reduces cell surface charge. This decrease in negative charge correlates with increased neutrophil adherence and aggregation, suggesting a role in inflammatory responses.

    Area of Science:

    • Immunology
    • Cell Biology
    • Biochemistry

    Background:

    • Neutrophils (polymorphonuclear leukocytes [PMN]) exhibit decreased negative surface charge upon exposure to chemotactic factors, a phenomenon speculated to influence their behavior in vivo.
    • PMN adherence and aggregation are known to be enhanced by the release of lysosomal enzymes during degranulation.

    Purpose of the Study:

    • To investigate the relationship between neutrophil degranulation, surface charge alterations, and changes in adherence and aggregation.
    • To determine the role of extracellular ions (calcium and magnesium) in mediating these surface charge changes.

    Main Methods:

    • Human peripheral blood neutrophils were isolated using Hypaque-Ficoll and dextran sedimentation.
    • Neutrophils were exposed to various secretagogues, including ionophore A23187, phorbol myristate acetate, concanavalin A, and chemotactic factors (C5a or f-met-leu-phe) with cytochalasin B.
    • Surface charge was quantified using a cytopherometer, and lysozyme release was measured.

    Main Results:

    • Secretagogue-induced degranulation led to a greater decrease in PMN surface charge compared to chemotactic factors alone.
    • The reduction in surface charge by f-met-leu-phe plus cytochalasin B required extracellular calcium and magnesium.
    • Changes in surface charge correlated with lysozyme release and increased cell adhesiveness to plastic surfaces.

    Conclusions:

    • Exocytosis of lysosomal granule contents is associated with a decrease in neutrophil surface charge.
    • A correlation exists between reduced surface charge and enhanced neutrophil aggregation and adhesiveness.
    • The precise causal relationship between degranulation, surface charge reduction, and aggregation requires further investigation.

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