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European Cytokine Network

, Volume 28, Issue 1, pp 19–26 | Cite as

Upregulation of cytokine mRNA in circulating leukocytes during human endotoxemia

  • Petra Jilma-Stohlawetz
  • Tuende Kliegel
  • Irene Kantner-Schlifke
  • Claudia Strasser-Marsik
  • Florian B. Mayr
  • Bernd Jilma
Research Article

Abstract

Background

Endotoxemia induces pronounced changes in leukocyte count and enhances the release of many cytokines. However, the molecular regulation of this cytokine release is poorly characterized in humans.

Methods

The time course of mRNA expression of 24 cytokines in circulating leukocytes was studied in a wellstandardized model of human endotoxemia (2 ng/kg). Real-time polymerase chain reaction (RT-PCR) was used to quantify the lipopolysaccharide (LPS)-inducible mRNA levels of leukocytes from 16 healthy volunteers in a randomized, placebo-controlled trial.

Results

Baseline mRNA levels of interleukins including IL-1α, IL-3, IL-5, IL-6, IL-12p40, IL-13, IL-15, IL-17, granulocyte colony-stimulating factor (G-CSF) and granulocyte monocyte CSF (GM-CSF) were below detectable levels in normal blood of the healthy participants. After 2 h, LPS infusion increased median mRNA levels of IL-1α by >1100-fold and IL-1β and IL-8 by 33-fold and 46-fold, respectively. In contrast, levels of tumor necrosis factor (TNF-α) and IL-10 mRNA increased by only 7-fold, whereas changes in mRNA expression of other cytokines showed either a more than two fold increase or were undetectable. In vitro incubation of whole blood with 50 pg/mL LPS for 2 h enhanced transcription levels of IL-1α mRNA by >10,000-fold, IL-6 and IL-12p40 by >1000-fold, IL-1α by 400-fold, TNF-α by 100-fold, IL-8, IL-18, interferon γ (IFN-γ) and G-CSF by >10-25-fold, and IL-10, IL-12p35, TNF-β, and IL-13 by 10-25-fold.

Conclusion

Only half of the 24 evaluated cytokines were expressed at the mRNA level in circulating leukocytes under basal conditions and after an LPS challenge. Only IL-1α, IL-1β, IL-10, IL-8, and TNF-α were upregulated in the circulating leukocytes, whereas several other cytokines (including IL-6 and G-CSF), were expressed on the mRNA level following in vitro incubation of blood with LPS. In addition, IL-1α and IL-1β might be potential diagnostic targets in inflammatory diseases.

Key words

randomized controlled trial endotoxemia mRNA cytokines RT-PCR 

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Copyright information

© John Libbey Eurotext 2017

Authors and Affiliations

  • Petra Jilma-Stohlawetz
    • 1
  • Tuende Kliegel
    • 2
  • Irene Kantner-Schlifke
    • 2
  • Claudia Strasser-Marsik
    • 2
  • Florian B. Mayr
    • 2
  • Bernd Jilma
    • 2
  1. 1.Dept. of Laboratory MedicineMedical University of ViennaViennaAustria
  2. 2.Department of Clinical Pharmacology, Division of Immunology and HematologyMedical University of ViennaWienAustria

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