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Published ahead of print on August 18, 2005, doi:10.1165/rcmb.2005-0181OC

Am. J. Respir. Cell Mol. Biol., Volume 33, Number 6, December 2005, 601-609

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Submitted on May 13, 2005
Revised on August 17, 2005

The P2Y14 Receptor of Airway Epithelial Cells: Coupling to Intracellular Ca2+ and IL-8 Secretion

Tobias Muller1, Hans Bayer1, Daniel Myrtek1, Davide Ferrari2, Stephan Sorichter1, Manfred W Ziegenhagen3, Gernot Zissel1, J. Christian Virchow Jr.4, Werner Luttmann4, Johannes Norgauer5, Francesco Di Virgilio2, and Marco Idzko1*

1 Department of Pneumology, University of Freiburg, Freiburg, Germany, 2 Department of Experimental and Diagnostic Medicine, Section of General Pathology and Interdisciplinary Center for the Study of Inflammation (ICSI), University of Ferrara, Ferrara, Italy, 3 Wilhelm-Anton-Hospital, Goch, Germany, 4 Department of Pneumology, University of Rostock, Rostock, Germany, 5 Department of Dermatology, University of Jena, Jena, Germany

* To whom correspondence should be addressed. E-mail: idzko{at}med1.ukl.uni-freiburg.de.

Uridine nucleotides and UDP-glucose are endogenous molecules, which are released into the extracellular environment in a lytic manner after cell damage, as well as by regulated nonlytic mechanisms. Recently, a UDP-glucose-specific Gi protein-coupled P2Y receptor, namely P2Y14, has been cloned. In this study, we demonstrated expression of the P2Y14 mRNA in human lung epithelial cells and in the epithelial cell lines A549 and BEAS-2B. Evidence of functional expression of the P2Y14 receptor in these cell lines was provided by calcium measurements after stimulation with uridine 5'-diphosphoglucose (UDP-glc). Experiments with pertussis toxin and the Ca2+-chelator EGTA revealed participation of pertussis-toxin sensitive Gi/o-proteins in the mobilization of Ca2+-ions from intracellular stores by UDP-glc. Moreover UDP-glc increased secretion of the potent neutrophil chemoattractant CXCL8/interleukin-8 (IL-8) in A549 and BEAS-2B cells in a pertussis toxin-sensitive manner. Moreover, reverse transcription and quantitative polymerase chain reaction revealed that UDP-glc modulated mRNA levels of IL-8/CXCL8. However, stimulation of A549 and BEAS-2B cells with UDP-glc neither modified basal nor cytokine-induced secretion of the CXC-chemokines CXCL9/MIG, CXCL10/IP-10, and CXCL11/I-TAC. In addition, UDP-glc did not affect proliferation of the two cell lines. In summary our data provide evidence for a distinct physiological role of P2Y14 in the selective release of specific chemokines from human airway epithelial cells.




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