Activation of the P2X7 receptor induces the rapid shedding of CD23 from human and murine B cells

Immunol Cell Biol. 2015 Jan;93(1):77-85. doi: 10.1038/icb.2014.69. Epub 2014 Aug 26.

Abstract

Activation of the P2X7 receptor by the extracellular damage-associated molecular pattern, adenosine 5'-triphosphate (ATP), induces the shedding of cell surface molecules including the low-affinity IgE receptor, CD23, from human leukocytes. A disintegrin and metalloprotease (ADAM) 10 mediates P2X7-induced shedding of CD23 from multiple myeloma RPMI 8226 B cells; however, whether this process occurs in primary B cells is unknown. The aim of the current study was to determine whether P2X7 activation induces the rapid shedding of CD23 from primary human and murine B cells. Flow cytometric and ELISA measurements showed that ATP treatment of human and murine B cells induced the rapid shedding of CD23. Treatment of cells with the specific P2X7 antagonist, AZ10606120, near-completely impaired ATP-induced CD23 shedding from both human and murine B cells. ATP-induced CD23 shedding was also impaired in B cells from P2X7 knockout mice. The absence of full-length, functional P2X7 in the P2X7 knockout mice was confirmed by immunoblotting of splenic cells, and by flow cytometric measurements of ATP-induced YO-PRO-1(2+) uptake into splenic B and T cells. The broad-spectrum metalloprotease antagonist, BB-94, and the ADAM10 antagonist, GI254023X, impaired P2X7-induced CD23 shedding from both human and murine B cells. These data indicate that P2X7 activation induces the rapid shedding of CD23 from primary human and murine B cells and that this process may be mediated by ADAM10.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / antagonists & inhibitors
  • ADAM Proteins / genetics
  • ADAM Proteins / metabolism*
  • ADAM10 Protein
  • Adenosine Triphosphate / pharmacology*
  • Amyloid Precursor Protein Secretases / antagonists & inhibitors
  • Amyloid Precursor Protein Secretases / genetics
  • Amyloid Precursor Protein Secretases / metabolism*
  • Animals
  • B-Lymphocytes / cytology
  • B-Lymphocytes / drug effects
  • B-Lymphocytes / metabolism*
  • Benzoxazoles
  • Dipeptides / pharmacology
  • Fluorescent Dyes
  • Gene Expression Regulation
  • Humans
  • Hydroxamic Acids / pharmacology
  • Lymphocyte Activation
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Mice, Knockout
  • Microscopy, Fluorescence
  • Phenylalanine / analogs & derivatives
  • Phenylalanine / pharmacology
  • Primary Cell Culture
  • Protease Inhibitors / pharmacology
  • Purinergic P2X Receptor Antagonists / pharmacology
  • Quinolinium Compounds
  • Receptors, IgE / genetics
  • Receptors, IgE / metabolism*
  • Receptors, Purinergic P2X7 / deficiency
  • Receptors, Purinergic P2X7 / genetics*
  • Signal Transduction
  • Spleen / cytology
  • Spleen / drug effects
  • Spleen / metabolism*
  • Thiophenes / pharmacology

Substances

  • 3-(formylhydroxyamino)-2-(3-phenyl-1-propyl)butanoic acid (2,2-dimethyl-1-methylcarbamoyl-1-propyl)amide
  • Benzoxazoles
  • Dipeptides
  • Fluorescent Dyes
  • Hydroxamic Acids
  • Membrane Proteins
  • Protease Inhibitors
  • Purinergic P2X Receptor Antagonists
  • Quinolinium Compounds
  • Receptors, IgE
  • Receptors, Purinergic P2X7
  • Thiophenes
  • YO-PRO 1
  • Phenylalanine
  • Adenosine Triphosphate
  • batimastat
  • Amyloid Precursor Protein Secretases
  • ADAM Proteins
  • ADAM10 Protein
  • ADAM10 protein, human