Skip to main content
SpringerLink
Log in

Upregulation of cytokine mRNA in circulating leukocytes during human endotoxemia

  • Research Article
  • Published:
European Cytokine Network Aims and scope

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Hochstein HD, Mills DF, Outschoorn AS, Rastogi SC. The processing and collaborative assay of a reference endotoxin. J Biol Stand 1983; 11:251.

    Article  CAS  PubMed  Google Scholar 

  2. Fiuza C, Suffredini AF. Human models of innate immunity: local and systemic inflammatory responses. J Endotoxin Res 2001; 7:385.

    Article  CAS  PubMed  Google Scholar 

  3. Martich GD, Boujoukos AJ, Suffredini AF. Response of man to endotoxin. Immunobiology 1993; 187:403.

    Article  CAS  PubMed  Google Scholar 

  4. Suffredini AF, Fantuzzi G, Badolato R, Oppenheim JJ, O’Grady NP. New insights into the biology of the acute phase response. J Clin Immunol 1999; 19:203.

    Article  CAS  PubMed  Google Scholar 

  5. Thaler B, Hohensinner PJ, Krychtiuk KA, et al. Differential in vivo activation of monocyte subsets during low-grade inflammation through experimental endotoxemia in humans. Sci Rep 2016; 6: 30162.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Leitner JM, Firbas C, Mayr FB, Reiter RA, Steinlechner B, Jilma B. Recombinant human antithrombin inhibits thrombin formation and interleukin 6 release in human endotoxemia. Clin Pharmacol Ther 2006; 79:23.

    Article  CAS  PubMed  Google Scholar 

  7. Suffredini AF, Noveck RJ. Human endotoxin administration as an experimental model in drug development. Clin Pharmacol Ther 2014; 96:418.

    Article  CAS  PubMed  Google Scholar 

  8. Spek CA, Verbon A, Aberson H, et al. Treatment with an anti-CD14 monoclonal antibody delays and inhibits lipopolysaccharide-induced gene expression in humans in vivo. J Clin Immunol 2003; 23:132.

    Article  CAS  PubMed  Google Scholar 

  9. Olszyna DP, Verbon A, Pribble JP, et al. Effect of IC14, an anti-CD14 antibody, on plasma and cell-associated chemokines during human endotoxemia. Eur Cytokine Netw 2003; 14:158.

    CAS  PubMed  Google Scholar 

  10. Jansen J, van der Poll T, Levi M, et al. Inhibition of the release of soluble tumor necrosis factor receptors in experimental endotoxemia by an anti-tumor necrosis factor-alpha antibody. J Clin Immunol 1995; 15:45.

    Article  CAS  PubMed  Google Scholar 

  11. Matzneller P, Strommer S, Drucker C, et al. Colistin reduces LPS-triggered inflammation in a human sepsis model in vivo: a randomized controlled trial. Clin Pharmacol Ther 2016. doi: 10.1002/cpt.582 [Epub ahead of print].

    Google Scholar 

  12. Kumar A, Zanotti S, Bunnell G, et al. Interleukin-10 blunts the human inflammatory response to lipopolysaccharide without affecting the cardiovascular response. Crit Care Med 2005; 33:331.

    Article  CAS  PubMed  Google Scholar 

  13. Pajkrt D, Manten A, van der Poll T, et al. Modulation of cytokine release and neutrophil function by granulocyte colonystimulating factor during endotoxemia in humans. Blood 1997; 90: 1415.

    CAS  PubMed  Google Scholar 

  14. Preas H.L. 2nd, Reda D, Tropea M, et al. Effects of recombinant soluble type I interleukin-1 receptor on human inflammatory responses to endotoxin. Blood 1996; 88: 2465.

    CAS  PubMed  Google Scholar 

  15. Derhaschnig U, Bergmair D, Marsik C, Schlifke I, Wijdenes J, Jilma B. Effect of interleukin-6 blockade on tissue factor-induced coagulation in human endotoxemia. Crit Care Med 2004; 32: 1136.

    Article  CAS  PubMed  Google Scholar 

  16. van den Blink B, Branger J, Weijer S, et al. P38 mitogen activated protein kinase is involved in the down regulation of granulocyte CXC chemokine receptors 1 and 2 during human endotoxemia. J Clin Immunol 2004; 24:37.

    Article  PubMed  Google Scholar 

  17. De Vries F, Leuschner J, Jilma B, Derhaschnig U. Establishment of a low dose canine endotoxemia model to test anti-inflammatory drugs: effects of prednisolone. Int J Immunopathol Pharmacol 2013; 26:861.

    Article  PubMed  Google Scholar 

  18. De Kleijn P, Odent T, Berntorp E, et al. Differences between developed and developing countries in paediatric care in haemophilia. Haemophilia 2012; 18:94.

    Article  PubMed  Google Scholar 

  19. Kamisoglu K, Calvano SE, Coyle SM, Corbett SA, Androulakis IP. Integrated transcriptional and metabolic profiling in human endotoxemia. Shock 2014; 42:499.

    Article  CAS  PubMed  Google Scholar 

  20. Haimovich B, Reddell MT, Calvano JE, et al. A novel model of common Toll-like receptor 4-and injury-induced transcriptional themes in human leukocytes. Crit Care 2010; 14: R177.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Xiao W, Mindrinos MN, Seok J, et al. A genomic storm in critically injured humans. J Exp Med 2011; 208: 2581.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Jabbari A, Suarez-Farinas M, Dewell S, Krueger JG. Transcriptional profiling of psoriasis using RNA-seq reveals previously unidentified differentially expressed genes. J Invest Dermatol 2012; 132:246.

    Article  CAS  PubMed  Google Scholar 

  23. McLoughlin K, Turteltaub K, Bankaitis-Davis D, et al. Limited dynamic range of immune response gene expression observed in healthy blood donors using RT-PCR. Mol Med 2006; 12:185.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Hollenstein U, Homoncik M, Knobl P, et al. Acenocoumarol decreases tissue factor-dependent coagulation during systemic inflammation in humans. Clin Pharmacol Ther 2002; 71:368.

    Article  CAS  PubMed  Google Scholar 

  25. Jilma B, Marsik C, Mayr F, et al. Pharmacodynamics of active siteinhibited factor VIIa in endotoxin-induced coagulation in humans. Clin Pharmacol Ther 2002; 72:403.

    Article  CAS  PubMed  Google Scholar 

  26. van Oostrom AJ, Sijmonsma TP, Verseyden C, et al. Postprandial recruitment of neutrophils may contribute to endothelial dysfunction. J Lipid Res 2003; 44:576.

    Article  PubMed  Google Scholar 

  27. Marsik C, Mayr F, Cardona F, Schaller G, Wagner OF, Jilma B. Endotoxin down-modulates P-selectin glycoprotein ligand-1 (PSGL-1 CD162) on neutrophils in humans. J Clin Immunol 2004; 24: 62–5.

    Article  CAS  PubMed  Google Scholar 

  28. Pahl A, Brune K. Stabilization of gene expression profiles in blood after phlebotomy. Clin Chem 2002; 48: 2251.

    CAS  PubMed  Google Scholar 

  29. Copeland S, Warren HS, Lowry SF, et al. Acute inflammatory response to endotoxin in mice and humans. Clin Diagn Lab Immunol 2005; 12:60.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Jilma B, Hergovich N, Homoncik M, et al. Granulocyte colony-stimulating factor (G-CSF) down regulates its receptor (CD114) on neutrophils and induces gelatinase B release in humans. Br J Haematol 2000; 111:314.

    Article  CAS  PubMed  Google Scholar 

  31. Hollenstein U, Homoncik M, Stohlawetz PJ, et al. Endotoxin down-modulates granulocyte colony-stimulating factor receptor (CD114) on human neutrophils. J Infect Dis 2000; 182:343.

    Article  CAS  PubMed  Google Scholar 

  32. Wagner JG, Roth RA. Neutrophil migration during endotoxemia. J Leukoc Biol 1999; 66:10.

    CAS  PubMed  Google Scholar 

  33. Fong YM, Marano MA, Moldawer LL, et al. The acute splanchnic and peripheral tissue metabolic response to endotoxin in humans. J Clin Invest 1990; 85: 1896.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Dekkers PE, Lauw FN, ten Hove T, et al. The effect of a metalloproteinase inhibitor (GI5402) on tumor necrosis factor-alpha (TNF-alpha) and TNF-alpha receptors during human endotoxemia. Blood 1999; 94: 2252.

    CAS  PubMed  Google Scholar 

  35. Watkins LR, Hansen MK, Nguyen KT, Lee JE, Maier SF. Dynamic regulation of the proinflammatory cytokine, interleukin-1beta: molecular biology for non-molecular biologists. Life Sci 1999; 65: 449.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dept. of Laboratory Medicine, Medical University of Vienna, Vienna, Austria

    Petra Jilma-Stohlawetz

  2. Department of Clinical Pharmacology, Division of Immunology and Hematology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090, Wien, Austria

    Tuende Kliegel, Irene Kantner-Schlifke, Claudia Strasser-Marsik, Florian B. Mayr & Bernd Jilma

Authors
  1. Petra Jilma-Stohlawetz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tuende Kliegel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Irene Kantner-Schlifke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Claudia Strasser-Marsik
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Florian B. Mayr
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bernd Jilma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bernd Jilma.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jilma-Stohlawetz, P., Kliegel, T., Kantner-Schlifke, I. et al. Upregulation of cytokine mRNA in circulating leukocytes during human endotoxemia. Eur Cytokine Netw 28, 19–26 (2017). https://doi.org/10.1684/ecn.2017.0389

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1684/ecn.2017.0389

Key words

Search

Navigation