Identification of cytokine-regulated genes in human leukocytes in vivo

J Allergy Clin Immunol. 2000 Apr;105(4):760-8. doi: 10.1067/mai.2000.104382.

Abstract

Background: Human polymorphic nuclear granulocytes (PMNs) such as neutrophils and eosinophils play a critical role in mediating inflammatory responses to microbial and parasitic infections. Exposure of these leukocytes to cytokines leads to an amplification of granulocyte effector functions by a mechanism termed "priming." Although many studies have investigated the effects of granulocyte priming, little is known concerning the molecular mechanisms that lead to this phenomenon.

Objective: The purpose of this study was to identify potential markers for granulocyte priming and thus also to gain further insight into the pathogenesis of inflammatory responses.

Methods: We used a modified differential display technique, random arbitrary primed-PCR to identify genes regulated during the priming of human polymorphic nuclear granulocytes by GM-CSF in vitro. Genes identified were validated by Northern blot analysis of in vitro and in vivo primed leukocytes.

Results: Several genes were identified and their expression characterized in vitro. One of these genes, 5-lipoxygenase-activating protein, was also found to be up-regulated in leukocytes isolated after allergen challenge of allergic asthmatic patients.

Conclusion: The use of differential display technology is a rapid and effective means of identifying genes whose expression is regulated by priming in vitro and in vivo. Further analysis will lead to a better understanding of the priming phenotype and may provide further insight into the pathologic mechanisms of inflammatory processes.

MeSH terms

  • 5-Lipoxygenase-Activating Proteins
  • Asthma / genetics*
  • Blotting, Northern
  • Carrier Proteins / blood
  • Carrier Proteins / genetics
  • Cytokines / blood
  • Cytokines / genetics*
  • DNA Primers
  • Gene Expression Regulation
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Inflammation / etiology
  • Inflammation / genetics*
  • Leukocytes / chemistry*
  • Membrane Proteins / blood
  • Membrane Proteins / genetics
  • Neutrophils / physiology
  • Polymerase Chain Reaction / methods
  • Tumor Necrosis Factor-alpha / pharmacology
  • Up-Regulation / drug effects

Substances

  • 5-Lipoxygenase-Activating Proteins
  • ALOX5AP protein, human
  • Carrier Proteins
  • Cytokines
  • DNA Primers
  • Membrane Proteins
  • Tumor Necrosis Factor-alpha
  • Granulocyte-Macrophage Colony-Stimulating Factor