title: Purinergic signaling on leukocytes infiltrating the LPS-injured lung.
abstract: Extracellular nucleotides and nucleosides have been implicated as important signaling molecules in the pathogenesis of acute lung injury (ALI). While adenosine is known to inhibit T cell activation, little information is available as to ATP and NAD degrading enzymes, the expression of ATP and adenosine receptors/transporters in different T cell subsets. ALI was induced by challenging mice with intra-tracheal instillation of 60 µl (3 µg/g) LPS. After 3 d and 7 d blood, lung tissue and bronchoalveolar lavage was collected and immune cells were analyzed using flow cytometry. The transcriptional phenotype of T helper cells, cytotoxic and regulatory T cells sorted by FACS was assessed by measuring the expression profile of 28 genes related to purinergic signaling using TaqMan Array Micro Fluidic Cards. Catabolism of ATP, NAD and cAMP by activated CD4+ T cells was evaluated by HPLC. CD73 was found to be highly abundant on lymphoid cells with little abundance on myeloid cells, while the opposite was true for CD39. After ALI, the abundance of CD39 and CD73 significantly increased on all T cell subsets derived from lung tissue and bronchoalveolar space. Expression analysis in T cell subsets of the lung revealed ATP (Cd39, Cd73) and NAD (Cd38, Cd157, Cd296, Pc-1) degrading enzymes. However, only transcription of Cd38, Cd39, Cd73, Ent1 and A2a receptor was significantly upregulated after ALI in T helper cells. CD4+ T cells from injured lung rapidly metabolized extracellular ATP to AMP and adenosine but not NAD or cAMP. These findings show that lung T cells--the dominant cell fraction in the later phase of ALI--exhibit a unique expression pattern of purinergic signaling molecules. Adenosine is formed by T cells at an enhanced rate from ATP but not from NAD and together with upregulated A2a receptor is likely to modulate the healing process after acute lung injury.
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Friebe
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany; Department of Anesthesiology and Intensive Care Medicine, Changhai Hospital, Second Military Medical University, Shanghai, PR China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Schmidt
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Nadine
authors.initials: N
authors.lastname: Borg
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Bodo
authors.initials: B
authors.lastname: Steckel
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Zhaoping
authors.initials: Z
authors.lastname: Ding
authors.affiliation: Department of Molecular Cardiology, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schrader
mesh_terms: D015720:5'-Nucleotidase / Q000276:immunology; D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D001081:Apyrase / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D007962:Leukocytes / Q000276:immunology / Q000473:pathology*; D008070:Lipopolysaccharides / Q000633:toxicity*; D055370:Lung Injury / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction; D011983:Receptors, Purinergic / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D015703:Antigens, CD; D008070:Lipopolysaccharides; D011983:Receptors, Purinergic; D015720:5'-Nucleotidase; D001081:Apyrase; C067869:CD39 antigen
references.citation: Glia. 2007 Apr 15;55(6):604-16
references.pmid: 17299767
references.citation: Physiology (Bethesda). 2009 Oct;24:298-306
references.pmid: 19815856
references.citation: N Engl J Med. 2000 May 4;342(18):1334-49
references.pmid: 10793167
references.citation: Purinergic Signal. 2006 Jun;2(2):409-30
references.pmid: 18404480
references.citation: Purinergic Signal. 2007 Mar;3(1-2):71-81
references.pmid: 18404420
references.citation: Biochem Biophys Res Commun. 2009 Jul 10;384(4):512-8
references.pmid: 19426712
references.citation: Br J Pharmacol. 2008 Mar;153(6):1331-40
references.pmid: 18157164
references.citation: Toxicol Sci. 2002 Dec;70(2):157-8
references.pmid: 12441359
references.citation: Eur J Immunol. 2011 May;41(5):1231-41
references.pmid: 21469131
references.citation: Microbes Infect. 2003 May;5(6):515-26
references.pmid: 12758281
references.citation: Nat Rev Drug Discov. 2008 Sep;7(9):759-70
references.pmid: 18758473
references.citation: FASEB J. 2009 Jun;23(6):1685-93
references.pmid: 19211924
references.citation: Circ Res. 2013 Jul 19;113(3):301-12
references.pmid: 23720442
references.citation: J Exp Med. 2007 Jun 11;204(6):1257-65
references.pmid: 17502665
references.citation: Int Immunol. 1999 Feb;11(2):179-89
references.pmid: 10069416
references.citation: FASEB J. 2009 Feb;23(2):473-82
references.pmid: 18838482
references.citation: Sci Signal. 2008 Sep 30;1(39):ra6
references.pmid: 18827222
references.citation: J Immunol. 2007 Jul 15;179(2):1254-63
references.pmid: 17617618
references.citation: J Immunol. 2011 Aug 1;187(3):1486-95
references.pmid: 21724990
references.citation: Oncoimmunology. 2013 Sep 1;2(9):e26246
references.pmid: 24319640
references.citation: PLoS One. 2012;7(2):e32416
references.pmid: 22389701
references.citation: J Clin Invest. 2002 Oct;110(7):993-1002
references.pmid: 12370277
references.citation: Immunity. 2003 Oct;19(4):571-82
references.pmid: 14563321
references.citation: J Clin Invest. 2009 Oct;119(10):2898-913
references.pmid: 19770521
references.citation: N Engl J Med. 2012 Dec 13;367(24):2322-33
references.pmid: 23234515
references.citation: Glia. 2013 Dec;61(12):2038-49
references.pmid: 24123515
references.citation: FASEB J. 2013 Aug;27(8):3078-89
references.pmid: 23603835
references.citation: J Chromatogr. 1990 May 18;527(2):414-20
references.pmid: 2387888
references.citation: Annu Rev Immunol. 2001;19:497-521
references.pmid: 11244045
references.citation: Blood. 2004 Feb 1;103(3):1124-30
references.pmid: 14525767
references.citation: N Engl J Med. 2005 Oct 20;353(16):1685-93
references.pmid: 16236739
references.citation: J Leukoc Biol. 2012 Oct;92(4):787-94
references.pmid: 22753954
references.citation: J Immunol. 2007 Jun 15;178(12):8127-37
references.pmid: 17548651
references.citation: Circ Res. 2006 Nov 10;99(10):1100-8
references.pmid: 17038639
references.citation: J Immunol. 1999 Jul 15;163(2):954-62
references.pmid: 10395692
references.citation: PLoS One. 2009;4(5):e5430
references.pmid: 19412549
references.citation: Am J Physiol Lung Cell Mol Physiol. 2005 Nov;289(5):L807-15
references.pmid: 15951336
references.citation: Blood. 2009 Jan 1;113(1):224-32
references.pmid: 18812468
references.citation: J Clin Invest. 2008 Oct;118(10):3301-15
references.pmid: 18787641
references.citation: PLoS Biol. 2013 Sep;11(9):e1001665
references.pmid: 24086109
references.citation: J Immunol. 2010 May 1;184(9):5271-9
references.pmid: 20348420
references.citation: Am J Respir Cell Mol Biol. 2012 Oct;47(4):417-26
references.pmid: 22721830
references.citation: J Immunol. 2006 Nov 15;177(10):6780-6
references.pmid: 17082591
references.citation: PLoS One. 2012;7(4):e34730
references.pmid: 22514659
references.citation: Anesth Analg. 2004 Feb;98(2):281-282
references.pmid: 14742354
references.citation: J Immunol. 2005 Mar 15;174(6):3298-305
references.pmid: 15749861
references.citation: FASEB J. 2013 Jun;27(6):2207-19
references.pmid: 23413361
references.citation: Am J Physiol Cell Physiol. 2011 Aug;301(2):C530-9
references.pmid: 21593451
references.citation: Biochem Soc Trans. 2007 Feb;35(Pt 1):109-14
references.pmid: 17233614
references.citation: Eur Respir J. 1999 Jan;13(1):66-70
references.pmid: 10836325