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Related Experiment Videos

A fast-acting elastase inhibitor in human monocytes.

E Remold-O'Donnell

    The Journal of Experimental Medicine
    |December 1, 1985
    PubMed
    Summary

    Human monocytes produce a potent elastase inhibitor that matures in culture. This fast-acting protein, distinct from alpha 1-proteinase inhibitor, regulates neutrophil and monocyte elastase activity at inflammatory sites.

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    Area of Science:

    • Biochemistry
    • Immunology
    • Cell Biology

    Background:

    • Neutrophil and pancreatic elastase are serine proteases implicated in various inflammatory conditions.
    • Elastase activity is regulated by endogenous proteinase inhibitors.
    • The role of monocyte-derived elastase inhibitors in inflammation is not fully understood.

    Purpose of the Study:

    • To detect and characterize a proteinase inhibitor active against neutrophil and pancreatic elastase in human monocytes and U937 cells.
    • To determine the kinetics, stability, and molecular properties of this monocyte-derived elastase inhibitor.
    • To investigate its potential role in regulating elastase activity at inflammatory sites.

    Main Methods:

    • Extraction and partial purification of proteinase inhibitors from cultured human monocytes and U937 cells.
    • Enzyme inhibition assays using neutrophil and pancreatic elastase.
    • Complex formation studies with 125I-labeled elastase, including stability in SDS and susceptibility to nucleophilic cleavage.
    • Electrophoretic analysis and immunological studies using antiserum to alpha 1-proteinase inhibitor.
    • Quantification of inhibitor activity and estimation of molecular weight.

    Main Results:

    • A novel elastase inhibitor was detected in cultured human monocytes and U937 cells, but not in freshly isolated monocytes.
    • The inhibitor forms a stable, covalent complex with elastase, is fast-acting (complete reaction < 1 min), and is distinct from alpha 1-proteinase inhibitor.
    • Inhibitor levels increase with monocyte maturation in culture, reaching maximum at 5-7 days.
    • The inhibitor is abundant in mature monocytes (≥ 1.5 X 10^6 molecules/cell) with an estimated molecular weight of 50,000.
    • The reaction with labeled elastase is proportional to inhibitor concentration, allowing for quantification.

    Conclusions:

    • Human monocytes and U937 cells synthesize and accumulate a potent, fast-acting elastase inhibitor during maturation.
    • This monocyte-derived inhibitor is biochemically distinct from plasma alpha 1-proteinase inhibitor.
    • Its high abundance, affinity for elastase, and rapid inhibition kinetics suggest a significant role in regulating neutrophil and monocyte elastase activity at inflammatory sites.

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