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Decrease in apparent Km for oxygen after stimulation of respiration of rat polymorphonuclear leukocytes.

S W Edwards, M B Hallett, D Lloyd

    FEBS Letters
    |September 5, 1983
    PubMed
    Summary
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    Polymorphonuclear leukocytes exhibit a two-phase respiratory burst upon stimulation. This cellular response shows a significant decrease in oxygen affinity during the second phase, suggesting oxygen concentration may regulate immune cell activity.

    Area of Science:

    • Immunology
    • Cellular Physiology
    • Biochemistry

    Background:

    • Polymorphonuclear leukocytes (PMNs) are critical immune cells involved in host defense.
    • The respiratory burst is a key function of PMNs, generating reactive oxygen species to combat pathogens.
    • Understanding the kinetics and regulation of the respiratory burst is essential for comprehending immune responses.

    Purpose of the Study:

    • To characterize the biphasic nature of the respiratory burst in PMNs.
    • To determine the kinetic parameters, specifically the apparent Km for oxygen, during different phases of PMN stimulation.
    • To explore the potential role of oxygen concentration in regulating PMN activity.

    Main Methods:

    • Stimulation of human PMNs using chemotactic peptide (N-formyl-methionyl-leucyl-phenylalanine) and cytochalasin B.

    Related Experiment Videos

  • Measurement of oxygen uptake to quantify the respiratory burst.
  • Calculation of apparent Km for oxygen for unstimulated and stimulated cells.

    • Main Results:

    • The respiratory burst in PMNs consists of two distinct phases: a rapid initial phase (1-3 min) and a slower, prolonged second phase.
    • The apparent Km for oxygen significantly decreased from 9.6 +/- 0.67 microM in unstimulated cells to 3.7 +/- 1.6 microM during the second phase of stimulation.
    • This reduction in Km indicates an increased affinity for oxygen during the second phase of the respiratory burst.

    Conclusions:

    • PMN respiratory burst is a biphasic process with distinct kinetic properties.
    • The decreased apparent Km for oxygen during stimulated PMN activity suggests a potential regulatory mechanism.
    • Low oxygen concentrations might influence PMN function in certain pathological conditions, warranting further investigation.