title: In vitro testing for anti-inflammatory properties of compounds employing peripheral blood mononuclear cells freshly isolated from healthy donors.
abstract: BACKGROUND
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Division of Biological Chemistry, Biocenter, Innsbruck Medical University, Fritz Pregl Strasse 3, 6020 Innsbruck, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Jenny
authors.forename: M
authors.initials: M
authors.lastname: Klieber
authors.forename: D
authors.initials: D
authors.lastname: Zaknun
authors.forename: S
authors.initials: S
authors.lastname: Schroecksnadel
authors.forename: K
authors.initials: K
authors.lastname: Kurz
authors.forename: M
authors.initials: M
authors.lastname: Ledochowski
authors.forename: H
authors.initials: H
authors.lastname: Schennach
authors.forename: Dietmar
authors.initials: D
authors.lastname: Fuchs
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D028421:Isoprostanes / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology; D008934:Mitogens / Q000494:pharmacology; D019798:Neopterin / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D053645:Interleukin-2 Receptor alpha Subunit; D028421:Isoprostanes; D008934:Mitogens; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha; D019798:Neopterin; D007371:Interferon-gamma; D014364:Tryptophan