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Related Experiment Videos

Endotoxin stimulates lymphocyte glutaminase expression

P Sarantos1, K Ockert, W W Souba

  • 1Department of Surgery, University of Florida College of Medicine, Gainesville.

Archives of Surgery (Chicago, Ill. : 1960)
|August 1, 1993
PubMed
Summary

Endotoxin increases glutaminase activity and gene expression in lymphocytes, providing essential fuel for cellular proliferation during infection. This molecular regulation supports immune responses when the gut barrier is compromised.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • Lymphocytes utilize glutamine as a primary fuel source.
  • Glutaminase enzyme activity regulates intracellular glutamine metabolism.
  • Lymphocyte glutamine utilization increases during infection to support proliferation.

Purpose of the Study:

  • To investigate the hypothesis that endotoxin regulates lymphocyte glutaminase expression at the molecular level.
  • To understand the impact of endotoxin on glutaminase activity and gene expression in lymphocytes.

Main Methods:

  • Adult rats were administered Escherichia coli endotoxin or saline.
  • Total RNA was extracted from lymphocytes for Northern hybridization using a glutaminase cDNA probe.
  • Glutaminase-specific activity and kinetic parameters were determined.

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  • Beta-actin mRNA served as a control for RNA loading.

    • Main Results:

    • Endotoxin administration increased glutaminase activity by 53% at 4 hours.
    • The increase in activity was primarily due to an 84% rise in Vmax.
    • Glutaminase mRNA levels showed a twofold increase at 2 hours and a fourfold increase at 4 hours.
    • The observed increase in mRNA preceded the increase in enzyme activity.

    Conclusions:

    • Endotoxin upregulates glutaminase expression and activity in lymphocytes.
    • This enhances glutamine availability for energy and proliferation in mesenteric lymph node lymphocytes.
    • This mechanism supports immune function during infection, particularly when the gut mucosal barrier is compromised.