Selectivity of recombinant human leukotriene D(4), leukotriene B(4), and lipoxin A(4) receptors with aspirin-triggered 15-epi-LXA(4) and regulation of vascular and inflammatory responses

Am J Pathol. 2001 Jan;158(1):3-9. doi: 10.1016/S0002-9440(10)63937-5.

Abstract

Aspirin-triggered lipoxin A(4) (ATL, 15-epi-LXA(4)) and leukotriene D(4) (LTD(4)) possess opposing vascular actions mediated via receptors distinct from the LXA(4) receptor (ALX) that is involved in leukocyte trafficking. Here, we identified these receptors by nucleotide sequencing and demonstrate that LTD(4) receptor (CysLT(1)) is induced in human vascular endothelia by interleukin-1beta. Recombinant CysLT(1) receptor gave stereospecific binding with both [(3)H]-LTD(4) and a novel labeled mimetic of ATL ([(3)H]-ATLa) that was displaced with LTD(4) and ATLa ( approximately IC(50) 0.2 to 0.9 nmol/L), but not with a bioinactive ATL isomer. The clinically used CysLT(1) receptor antagonist, Singulair, showed a lower rank order for competition with [(3)H]-ATLa (IC(50) approximately 8.3 nmol/L). In contrast, LTD(4) was an ineffective competitive ligand for recombinant ALX receptor with [(3)H]-ATLa, and ATLa did not compete for [(3)H]-LTB(4) binding with recombinant LTB(4) receptor. Endogenous murine CysLT(1) receptors also gave specific [(3)H]-ATLa binding that was displaced with essentially equal affinity by LTD(4) or ATLa. Systemic ATLa proved to be a potent inhibitor (>50%) of CysLT(1)-mediated vascular leakage in murine skin (200 microg/kg) in addition to its ability to block polymorphonuclear leukocyte recruitment to dorsal air pouch (4 microg/kg). These results indicate that ATL and LTD(4) bind and compete with equal affinity at CysLT(1), providing a molecular basis for aspirin-triggered LXs serving as a local damper of both vascular CysLT(1) signals as well as ALX receptor-regulated polymorphonuclear leukocyte traffic.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aspirin / pharmacology
  • Binding, Competitive / drug effects
  • COS Cells
  • Capillary Permeability / drug effects*
  • Cell Line
  • Dose-Response Relationship, Drug
  • Gene Expression
  • Humans
  • Hydroxyeicosatetraenoic Acids / chemistry
  • Hydroxyeicosatetraenoic Acids / metabolism
  • Hydroxyeicosatetraenoic Acids / pharmacology*
  • Inflammation / pathology
  • Inflammation / prevention & control*
  • Leukotriene D4 / chemistry
  • Leukotriene D4 / metabolism
  • Leukotriene D4 / pharmacology
  • Lipoxins*
  • Male
  • Membrane Proteins*
  • Mice
  • Mice, Inbred BALB C
  • Neutrophils / drug effects
  • Neutrophils / pathology
  • RNA / genetics
  • RNA / metabolism
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Receptors, Formyl Peptide*
  • Receptors, Leukotriene / genetics
  • Receptors, Leukotriene / metabolism
  • Receptors, Leukotriene B4 / genetics
  • Receptors, Leukotriene B4 / metabolism
  • Receptors, Lipoxin*
  • Recombinant Proteins / metabolism
  • Stereoisomerism
  • Tritium

Substances

  • FPR2 protein, human
  • Hydroxyeicosatetraenoic Acids
  • Lipoxins
  • Membrane Proteins
  • Receptors, Cell Surface
  • Receptors, Formyl Peptide
  • Receptors, Leukotriene
  • Receptors, Leukotriene B4
  • Receptors, Lipoxin
  • Recombinant Proteins
  • lipoxin A4
  • Tritium
  • RNA
  • Leukotriene D4
  • cysteinyl leukotriene receptor 2
  • leukotriene D4 receptor
  • Aspirin