Characterization of prostaglandin (PG)-binding sites expressed on human basophils. Evidence for a prostaglandin E1, I2, and a D2 receptor

J Biol Chem. 1992 Jun 25;267(18):12700-8.

Abstract

Recent data suggest that prostaglandins (PGs) are involved in the regulation of basophil activation. The aim of this study was to characterize the basophil PG-binding sites by means of radioreceptor assays using 3H-labeled PGs. Scatchard analysis for pure (greater than 95%) chronic myeloid leukemia (CML) basophils revealed two classes of PGE1-binding sites differing in their affinity for the natural ligand (Bmax1 = 217 +/- 65 fmol/10(8) cells; Kd1 = 0.5 +/- 0.2 nM; Bmax2 = 2462 +/- 381 fmol/10(8) cells; Kd2 = 47 +/- 20 nM; IC50 = PGE1 less than PGI2 less than PGD2 less than PGE2 less than PGF2 alpha) as well as two classes of PGI2 (iloprost)-binding sites (Bmax1 = 324 +/- 145 fmol/10(8) cells; Kd1 = 0.5 +/- 0.3 nM; Bmax2 = 2541 +/- 381; Kd2 = 27 +/- 6 nM; IC50 = PGI2 less than PGE1 less than PGD2 less than PGE2 less than PGF2 alpha. In addition, CML basophils exhibited a single class of PGD2-binding sites (Bmax = 378 +/- 98 fmol/10(8) cells; Kd = 13 +/- 4 nM; IC50: PGD2 less than PGI2 less than PGE1 less than PGE2 less than PGF2 alpha). In contrast, we were unable to detect specific saturable PGE2-binding sites. Primary and immortalized (KU812) CML basophils revealed an identical pattern of PG receptor expression. Basophils (KU812) expressed significantly (p less than 0.001) lower number of PGE1 (PGI2)-binding sites (Bmax1: 9% (20%) of control; Bmax2: 36% (50%) of control) when cultured with recombinant interleukin 3 (rhIL-3), a basophil-activating cytokine, whereas rhIL-2 had no effect on PG receptor expression. Functional significance of binding of PGs to basophils was provided by the demonstration of a dose-dependent increase in cellular cAMP upon agonist activation, with PGE1 (ED50 = 1.7 +/- 1.1 nM) and PGI2 (ED50 = 2.8 +/- 2.3 nM) being the most potent compounds. These findings suggest that human basophils express specific receptors for PGE1, PGI2 as well as for PGD2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alprostadil / metabolism
  • Basophils / metabolism*
  • Binding Sites
  • Cell Line
  • Cyclic AMP / metabolism
  • Epoprostenol / metabolism
  • Humans
  • Iloprost / metabolism
  • Interleukin-3 / physiology
  • Interleukins / physiology
  • Kinetics
  • Prostaglandin D2 / metabolism
  • Prostaglandins / metabolism*
  • Radioimmunoassay
  • Receptors, Epoprostenol
  • Receptors, Immunologic*
  • Receptors, Prostaglandin / metabolism*
  • Receptors, Prostaglandin E

Substances

  • Interleukin-3
  • Interleukins
  • Prostaglandins
  • Receptors, Epoprostenol
  • Receptors, Immunologic
  • Receptors, Prostaglandin
  • Receptors, Prostaglandin E
  • Epoprostenol
  • Cyclic AMP
  • Alprostadil
  • Iloprost
  • Prostaglandin D2
  • prostaglandin D2 receptor