Differential effects of cell density on 5-lipoxygenase (5-LO), five-lipoxygenase-activating protein (FLAP) and interleukin-1 beta (IL-1 beta) expression in human neutrophils

Inflammation. 1997 Apr;21(2):235-50. doi: 10.1023/a:1027326405788.

Abstract

We have analyzed the effect of cellular density of 5-Lipoxygenase (5-LO), 5-lipoxygenase-activating protein (FLAP) and interleukin-1 beta (IL-1 beta) gene expression in neutrophils from healthy subjects under culture conditions of low and high cell density. By using RT-PCR techniques, we have found that 5-LO mRNA accumulation decreased in cells cultured at high density, while FLAP mRNA is not affected. De novo 5-LO synthesis, as well as steady-state levels, were reduced in cells maintained at high density. In contrast, the high density conditions lead to the induction of IL-1 beta gene at the RNA and protein levels as measured by RT-PCR and by immunoprecipitation. These results suggest that cellular density plays a role in gene modulation when neutrophils are accumulating at an inflammatory site since neutrophils obtained from the synovial fluid of patients with RA exhibit a protein synthesis profile similar to that observed in peripheral blood neutrophils cultured at high density.

MeSH terms

  • 5-Lipoxygenase-Activating Proteins
  • Apoptosis
  • Arachidonate 5-Lipoxygenase / genetics
  • Arachidonate 5-Lipoxygenase / metabolism*
  • Arthritis, Rheumatoid / metabolism
  • Arthritis, Rheumatoid / pathology
  • Base Sequence
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cell Count
  • DNA Primers / genetics
  • Gene Expression
  • Humans
  • In Vitro Techniques
  • Inflammation Mediators / metabolism
  • Interleukin-1 / genetics
  • Interleukin-1 / metabolism*
  • Kinetics
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Neutrophils / cytology*
  • Neutrophils / metabolism*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism

Substances

  • 5-Lipoxygenase-Activating Proteins
  • ALOX5AP protein, human
  • Carrier Proteins
  • DNA Primers
  • Inflammation Mediators
  • Interleukin-1
  • Membrane Proteins
  • RNA, Messenger
  • Arachidonate 5-Lipoxygenase