The influence of glycosaminoglycans on IL-8-mediated functions of neutrophils

Carbohydr Res. 2012 Jul 15;356:196-203. doi: 10.1016/j.carres.2012.02.025. Epub 2012 Mar 5.

Abstract

Glycosaminoglycans (GAGs) of the extracellular matrix (ECM) contribute to the regulation of physiological processes by binding various immune-competent proteins. Due to their large structural diversity, the analysis of the binding properties and their functional consequences is challenging. The cytokine interleukin-8 (IL-8) is involved in the recruitment of neutrophils to inflammatory sites. Here, we investigated the interaction of heparin hexasaccharides and recombinant human IL-8, consisting of 77 amino acids using fluorescence and NMR spectroscopy. A dissociation constant of 2.0±0.4 μM was determined for the heparin-IL-8 complex, which is slightly higher than what has been found for chondroitin-6-sulfate (K(D)=1.4±0.4 μM) [Pichert, A.; Samsonov, S. A.; Theisgen, S.; Thomas, L.; Baumann, L.; Schiller, J.; Beck-Sickinger, A. G.; Huster, D.; Pisabarro, M. T. Glycobiology2012, 22, 134-145], suggesting an important role of the sulfate group at position 6 of the second ring in the disaccharide unit of the GAGs in this interaction. In addition, the influence of long-chain hyaluronan, chondroitin sulfate, and heparin on IL-8-induced chemotaxis and oxidative activity of neutrophils was examined. Only the incubation of heparin with IL-8 affected the IL-8-mediated chemotaxis of neutrophils. However, all investigated GAGs enhanced the IL-8-induced formation of reactive oxygen species in neutrophils, which is an entirely new finding. This work provides a representative example of how protein functions can be regulated by different GAGs of the ECM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cells, Cultured
  • Chemotaxis / drug effects
  • Chondroitin Sulfates / chemistry*
  • Chondroitin Sulfates / pharmacology
  • Dose-Response Relationship, Drug
  • Extracellular Matrix / chemistry
  • Heparin / chemistry*
  • Heparin / pharmacology
  • Humans
  • Hyaluronic Acid / chemistry*
  • Hyaluronic Acid / pharmacology
  • Interleukin-8 / chemistry
  • Interleukin-8 / genetics
  • Interleukin-8 / pharmacology*
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Neutrophils / cytology
  • Neutrophils / drug effects*
  • Neutrophils / metabolism
  • Protein Binding
  • Reactive Oxygen Species / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / pharmacology
  • Spectrometry, Fluorescence
  • Structure-Activity Relationship

Substances

  • Interleukin-8
  • Reactive Oxygen Species
  • Recombinant Proteins
  • Hyaluronic Acid
  • Heparin
  • Chondroitin Sulfates