title: Activation of the P2X7 receptor induces the rapid shedding of CD23 from human and murine B cells.
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.
authors.affiliation: 1] School of Biological Sciences, University of Wollongong, Wollongong, New South Wales, Australia [2] Illawarra Health and Medical Research Institute, Wollongong, New South Wales, Australia.
authors.forename: Aleta
authors.initials: A
authors.lastname: Pupovac
authors.affiliation: 1] School of Biological Sciences, University of Wollongong, Wollongong, New South Wales, Australia [2] Illawarra Health and Medical Research Institute, Wollongong, New South Wales, Australia.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Geraghty
authors.affiliation: 1] School of Biological Sciences, University of Wollongong, Wollongong, New South Wales, Australia [2] Illawarra Health and Medical Research Institute, Wollongong, New South Wales, Australia.
authors.forename: Debbie
authors.initials: D
authors.lastname: Watson
authors.affiliation: 1] School of Biological Sciences, University of Wollongong, Wollongong, New South Wales, Australia [2] Illawarra Health and Medical Research Institute, Wollongong, New South Wales, Australia.
authors.forename: Ronald
authors.initials: R
authors.lastname: Sluyter
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000072197:ADAM10 Protein; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D001583:Benzoxazoles; D004151:Dipeptides / Q000494:pharmacology; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000494:pharmacology; D061251:Primary Cell Culture; D011480:Protease Inhibitors / Q000494:pharmacology; D058920:Purinergic P2X Receptor Antagonists / Q000494:pharmacology; D011806:Quinolinium Compounds; D017455:Receptors, IgE / Q000235:genetics / Q000378:metabolism*; D058486:Receptors, Purinergic P2X7 / Q000172:deficiency / Q000235:genetics*; D015398:Signal Transduction; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013876:Thiophenes / Q000494:pharmacology
chemical_list: C555398:3-(formylhydroxyamino)-2-(3-phenyl-1-propyl)butanoic acid (2,2-dimethyl-1-methylcarbamoyl-1-propyl)amide; D001583:Benzoxazoles; D004151:Dipeptides; D005456:Fluorescent Dyes; D006877:Hydroxamic Acids; D008565:Membrane Proteins; D011480:Protease Inhibitors; D058920:Purinergic P2X Receptor Antagonists; D011806:Quinolinium Compounds; D017455:Receptors, IgE; D058486:Receptors, Purinergic P2X7; D013876:Thiophenes; C089813:YO-PRO 1; D010649:Phenylalanine; D000255:Adenosine Triphosphate; C080985:batimastat; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human