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Related Experiment Videos

Singlet oxygen production by human eosinophils.

J R Kanofsky1, H Hoogland, R Wever

  • 1Medical Service, Edward Hines, Jr. Veterans Administration Hospital, Hines, Illinois 60141.

The Journal of Biological Chemistry
|July 15, 1988
PubMed
Summary

Human eosinophils can generate singlet oxygen, a reactive oxygen species, in the presence of bromide ions. This groundbreaking discovery shows living cells producing singlet oxygen via a peroxidase-catalyzed mechanism.

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Area of Science:

  • Immunology
  • Biochemistry
  • Cellular Biology

Background:

  • Eosinophils are immune cells involved in inflammatory responses.
  • Reactive oxygen species (ROS) play crucial roles in cellular signaling and defense.
  • Singlet oxygen (¹O₂) is a highly reactive form of oxygen with potential biological implications.

Purpose of the Study:

  • To investigate the potential of human eosinophils to produce singlet oxygen.
  • To identify the conditions and mechanisms underlying singlet oxygen generation by eosinophils.
  • To compare the singlet oxygen-producing capacity of eosinophil peroxidase with other related enzymes.

Main Methods:

  • Human eosinophils were stimulated with phorbol myristate acetate (PMA).
  • Chemiluminescence at 1268 nm was measured to detect singlet oxygen production.

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  • Experiments were conducted with varying bromide ion concentrations.
  • Purified eosinophil peroxidase was used in a cell-free system with hydrogen peroxide and bromide.

    • Main Results:

    • Human eosinophils stimulated with PMA produced 1268 nm chemiluminescence indicative of singlet oxygen.
    • Singlet oxygen generation by eosinophils was dependent on the presence of bromide ions.
    • A bromide concentration of 100 microM was sufficient for measurable singlet oxygen production.
    • Eosinophil peroxidase demonstrated significantly higher singlet oxygen yield compared to myeloperoxidase and lactoperoxidase under identical conditions.
    • The cells remained viable after singlet oxygen production.

    Conclusions:

    • This study provides the first evidence of singlet oxygen production by living human eosinophils.
    • The generation of singlet oxygen by eosinophils is likely mediated by a peroxidase-catalyzed reaction involving bromide ions.
    • Eosinophil peroxidase exhibits a unique and potent ability to generate singlet oxygen compared to other peroxidases.