Lack of activity of 15-epi-lipoxin A₄ on FPR2/ALX and CysLT1 receptors in interleukin-8-driven human neutrophil function

Clin Exp Immunol. 2013 Aug;173(2):298-309. doi: 10.1111/cei.12110.

Abstract

Neutrophil recruitment and survival are important control points in the development and resolution of inflammatory processes. 15-epi-lipoxin (LX)A interaction with formyl peptide receptor 2 (FPR2)/ALX receptor is suggested to enhance anti-inflammatory neutrophil functions and mediate resolution of airway inflammation. However, it has been reported that 15-epi-LXA₄ analogues can also bind to cysteinyl leukotriene receptor 1 (CysLT1) and that the CysLT1 antagonist MK-571 binds to FPR2/ALX, so cross-reactivity between FPR2/ALX and CysLT1 ligands cannot be discarded. It is not well established whether the resolution properties reported for 15-epi-LXA4 are mediated through FPR2/ALX, or if other receptors such as CysLT1 may also be involved. Evaluation of specific FPR2/ALX ligands and CysLT1 antagonists in functional biochemical and cellular assays were performed to establish a role for both receptors in 15-epi-LXA₄-mediated signalling and function. In our study, a FPR2/ALX synthetic peptide (WKYMVm) and a small molecule FPR2/ALX agonist (compound 43) induced FPR2/ALX-mediated signalling, enhancing guanosine triphosphate-gamma (GTPγ) binding and decreasing cyclic adenosine monophosphate (cAMP) levels, whereas 15-epi-LXA₄ was inactive. Furthermore, 15-epi-LXA4 showed neither binding affinity nor signalling towards CysLT1. In neutrophils, 15-epi-LXA₄ showed a moderate reduction of interleukin (IL)-8-mediated neutrophil chemotaxis but no effect on neutrophil survival was observed. In addition, CysLT1 antagonists were inactive in FPR2/ALX signalling or neutrophil assays. In conclusion, 15-epi-LXA₄ is not a functional agonist or an antagonist of FPR2/ALX or CysLT1, shows no effect on IL-8-induced neutrophil survival and produces only moderate inhibition in IL-8-mediated neutrophil migration. Our data do not support an anti-inflammatory role of 15-epi-LXA₄- FPR2/ALX interaction in IL-8-induced neutrophil inflammation.

Keywords: 15-epi-LXA4; FPR2/ALX; apoptosis; interleukin-8; neutrophils.

MeSH terms

  • Cell Movement / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Humans
  • Interleukin-8 / immunology
  • Lipoxins / pharmacology*
  • Neutrophil Activation* / drug effects
  • Neutrophils / drug effects*
  • Neutrophils / immunology
  • Protein Binding / drug effects
  • Receptors, Formyl Peptide / agonists*
  • Receptors, Leukotriene / metabolism*
  • Receptors, Lipoxin / agonists*
  • Signal Transduction / drug effects

Substances

  • FPR2 protein, human
  • Interleukin-8
  • Lipoxins
  • Receptors, Formyl Peptide
  • Receptors, Leukotriene
  • Receptors, Lipoxin
  • lipoxin A4
  • Cyclic AMP
  • leukotriene D4 receptor