Neuromedin U elicits cytokine release in murine Th2-type T cell clone D10.G4.1.
Neuromedin U (NmU), originally isolated from porcine spinal cord and later from other species, is a novel peptide that potently contracts smooth muscle. NmU interacts with two G protein-coupled receptors designated as NmU-1R and NmU-2R. This study demonstrates a potential proinflammatory role for NmU. In a mouse Th2 cell line (D10.G4.1), a single class of high affinity saturable binding sites for (125)I-labeled NmU (K(D) 364 pM and B(max) 1114 fmol/mg protein) was identified, and mRNA encoding NmU-1R, but not NmU-2R, was present. Competition binding analysis revealed equipotent, high affinity binding of NmU isopeptides to membranes prepared from D10.G4.1 cells. Exposure of these cells to NmU isopeptides resulted in an increase in intracellular Ca(2+) concentration (EC(50) 4.8 nM for human NmU). In addition, NmU also significantly increased the synthesis and release of cytokines including IL-4, IL-5, IL-6, IL-10, and IL-13. Studies using pharmacological inhibitors indicated that maximal NmU-evoked cytokine release required functional phospholipase C, calcineurin, MEK, and PI3K pathways. These data suggest a role for NmU in inflammation by stimulating cytokine production by T cells.
Johnson
Eric N
EN
Department of High Throughput Biology, GlaxoSmithKline, Research Triangle Park, NC 27709, USA. eric.n.johnson@gsk.com
Appelbaum
Edward R
ER
Carpenter
Donald C
DC
Cox
Richard F
RF
Disa
Jyoti
J
Foley
James J
JJ
Ghosh
Sujoy K
SK
Naselsky
Diane P
DP
Pullen
Mark A
MA
Sarau
Henry M
HM
Scheff
Samuel R
SR
Steplewski
Klaudia M
KM
Zaks-Zilberman
Meirav
M
Aiyar
Nambi
N
D000818:Animals; D019703:Calcineurin / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002999:Clone Cells; D016207:Cytokines / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004285:Dogs; D004963:Estrenes / Q000494:pharmacology; D006801:Humans; D007378:Interleukins / Q000037:antagonists & inhibitors / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000276:immunology; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism / Q000502:physiology*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology; D011760:Pyrrolidinones / Q000494:pharmacology; D051381:Rats; D019948:Receptors, Interleukin-4 / Q000502:physiology; D017981:Receptors, Neurotransmitter / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000276:immunology; D013552:Swine; D018418:Th2 Cells / Q000276:immunology* / Q000378:metabolism*; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000502:physiology
D016207:Cytokines; D004963:Estrenes; D007378:Interleukins; D008565:Membrane Proteins; D009479:Neuropeptides; D011760:Pyrrolidinones; D019948:Receptors, Interleukin-4; D017981:Receptors, Neurotransmitter; C082444:neuromedin U receptor; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C056900:neuromedin U; D019869:Phosphatidylinositol 3-Kinases; D020928:Mitogen-Activated Protein Kinases; D019703:Calcineurin; D010738:Type C Phospholipases; D002118:Calcium