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In vitro testing for anti-inflammatory properties of compounds employing peripheral blood mononuclear cells freshly isolated from healthy donors

Abstract

Introduction

Inflammation is crucially involved in a variety of diseases like autoimmune syndromes, cardiovascular and neurodegenerative disorders, cancer, sepsis and allograft rejection.

Methods

Freshly isolated human peripheral blood mononuclear cells (PBMCs) are used as a screening assay for anti-inflammatory properties of compounds. Determinations of neopterin production by ELISA and of tryptophan degradation by HPLC are used as read-outs. Results are compared with further markers of immune response and oxidative stress.

Results

Phytohaemagglutinin induced significant tryptophan degradation and neopterin formation in PBMC, which correlated with IFN-γ, TNF-α, soluble cytokine receptors and isoprostane-8. Addition of vitamin C and E suppressed the responses dose-dependently.

Discussion

The determination of tryptophan degradation and neopterin production in PBMC reflects various pro- and anti-inflammatory cascades that are of relevance also in patients. It constitutes a robust and reliable approach to screen anti-inflammatory or immunosuppressive drugs and may improve throughput, speed and cost-effectiveness in drug discovery.

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Acknowledgments

The authors thank Miss Maria Gleinser for excellent technical assistance.

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Correspondence to Dietmar Fuchs.

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Responsible Editor: Liwu Li.

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Jenny, M., Klieber, M., Zaknun, D. et al. In vitro testing for anti-inflammatory properties of compounds employing peripheral blood mononuclear cells freshly isolated from healthy donors. Inflamm. Res. 60, 127–135 (2011). https://doi.org/10.1007/s00011-010-0244-y

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  • DOI: https://doi.org/10.1007/s00011-010-0244-y

Keywords

  • Tryptophan
  • Kynurenine
  • Indoleamine 2
  • 3-Dioxygenase
  • Neopterin
  • Cytokine
  • Cytokine receptors
  • Interferon-γ