Incomplete activation of human eosinophils via the histamine H4-receptor: evidence for ligand-specific receptor conformations

Biochem Pharmacol. 2012 Jul 15;84(2):192-203. doi: 10.1016/j.bcp.2012.04.004. Epub 2012 Apr 11.

Abstract

Eosinophils play a crucial role in the pathogenesis of allergic diseases. Histamine activates eosinophils via the H(4)-receptor (H(4)R). However, pharmacological analysis of the H(4)R in eosinophils is still incomplete, and cell purity is a problem. The H(4)R antagonist 1-[(5-chloro-1H-indol-2-yl)carbonyl]-4-methylpiperazine (JNJ7777120) has recently been reported to exhibit paradoxical stimulatory effects in some systems. Therefore, the first aim of our study was to pharmacologically re-examine H(x)R subtypes on human eosinophils using a highly purified preparation (97±2%). The second aim was to compare the effects of histamine with those induced by well-known activators of eosinophil functions, i.e. eotaxin-1 and formyl peptides. Histamine and the H(4)R-selective agonist 2-cyano-1-[4-(1H-imidazol-4-yl)butyl]-3-[(2-phenylthio)ethyl]guanidine (UR-PI376) increased intracellular calcium concentration ([Ca(2+)](i)) and activated chemotaxis. JNJ7777120 per se exhibited no stimulatory effects but inhibited stimulation by histamine and UR-PI376. Blockade of the H(2)R by famotidine enhanced histamine-induced chemotaxis but not rises in [Ca(2+)](i). Compared to eotaxin and formyl peptides, the effect of histamine on eosinophil chemotaxis was only small. Formyl peptides but not histamine activated reactive oxygen species formation and release of eosinophil peroxidase. In conclusion, histamine is only an incomplete eosinophil activator with the H(2)R blunting the small chemotactic response to H(4)R activation. We also noted several differences in potencies of histamine, UR-PI376 and JNJ7777120 in calcium and chemotaxis assays and when compared to results in the literature. This indicates functional selectivity of H(4)R ligands, thus ligand-specific stabilization of distinct receptor conformations, inducing distinct biological responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism
  • Cells, Cultured
  • Chemokine CCL11 / metabolism
  • Chemokine CCL11 / pharmacology
  • Chemotaxis / drug effects
  • Eosinophil Peroxidase / metabolism
  • Eosinophils / drug effects
  • Eosinophils / physiology*
  • Guanidines / pharmacology
  • Humans
  • Imidazoles / pharmacology
  • Indoles / pharmacology
  • Ligands
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
  • Piperazines / pharmacology
  • Reactive Oxygen Species / metabolism
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / antagonists & inhibitors
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Histamine / metabolism*
  • Receptors, Histamine H4

Substances

  • 2-cyano-1-(4-(1H-imidazol-4-yl)butyl)-3-((2-phenylthio)ethyl)guanidine
  • Chemokine CCL11
  • Guanidines
  • HRH4 protein, human
  • Imidazoles
  • Indoles
  • Ligands
  • Piperazines
  • Reactive Oxygen Species
  • Receptors, G-Protein-Coupled
  • Receptors, Histamine
  • Receptors, Histamine H4
  • 1-((5-chloro-1H-indol-2-yl)carbonyl)-4-methylpiperazine
  • N-Formylmethionine Leucyl-Phenylalanine
  • Eosinophil Peroxidase
  • Calcium