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Thromb Haemost 2014; 112(06): 1230-1243
DOI: 10.1160/th14-04-0312
Wound Healing and Inflammation/Infection
Schattauer GmbH

Targeted mass spectrometry analysis of neutrophil-derived proteins released during sepsis progression

Erik Malmström
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
,
Alzbeta Davidova
2   Department of Infectious and Tropical Diseases, First Faculty of Medicine, Charles University in Prague and Na Bulovce Hospital, Prague, Czech Republic
3   Department of Infectious Diseases, First Faculty of Medicine, Military University Hospital and Charles University in Prague, Prague, Czech Republic
,
Matthias Mörgelin
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
,
Adam Linder
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
4   Centre for Heart Lung Innovation, St. Paul’s Hospital, The University of British Columbia (UBC), Vancouver, British Columbia, Canada
,
Michael Larsen
5   Department of Biomedical Sciences, CFIM, The Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
,
Klaus Qvortrup
5   Department of Biomedical Sciences, CFIM, The Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
,
Pontus Nordenfelt
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
6   Springer Laboratory, Program in Cellular and Molecular Medicine, Children’s Hospital Boston, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA
,
Oonagh Shannon
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
,
Olga Dzupova
7   Department of Infectious Diseases, Third Faculty of Medicine, Charles University in Prague and Na Bulovce Hospital, Prague, Czech Republic
,
Michal Holub
2   Department of Infectious and Tropical Diseases, First Faculty of Medicine, Charles University in Prague and Na Bulovce Hospital, Prague, Czech Republic
3   Department of Infectious Diseases, First Faculty of Medicine, Military University Hospital and Charles University in Prague, Prague, Czech Republic
,
Johan Malmström
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
,
Heiko Herwald
1   Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, Lund University, Lund, Sweden
› Author Affiliations
Further Information

Publication History

Received: 04 April 2014

Accepted after major revision: 16 June 2014

Publication Date:
18 November 2017 (online)

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Summary

Early diagnosis of severe infectious diseases is essential for timely implementation of lifesaving therapies. In a search for novel biomarkers in sepsis diagnosis we focused on polymorphonuclear neutrophils (PMNs). Notably, PMNs have their protein cargo readily stored in granules and following systemic stimulation, an immediate increase of neutrophil-borne proteins can be observed into the circulation of sepsis patients. We applied a combination of mass spectrometry (MS) based approaches, LC-MS/MS and selected reaction monitoring (SRM), to characterise and quantify the neutrophil proteome in healthy or disease conditions. With this approach we identified a neutrophil- derived protein abundance pattern in blood plasma consisting of 20 proteins that can be used as a protein signature for severe infectious diseases. Our results also show that SRM is highly sensitive, specific, and reproducible and, thus, a promising technology to study a complex, dynamic and multifactorial disease such as sepsis.

Keywords

Coagulation - infectious diseases - innate immunity - neutrophils - mass spectrometry - proteomics

 

  • References

  • 1 Dombrovskiy VY, Martin AA, Sunderram J. et al. Rapid increase in hospitalisation and mortality rates for severe sepsis in the United States: A trend analysis from 1993 to 2003. Crit Care Med 2007; 35: 1244-1250.
    CrossrefPubMedGoogle Scholar
  • 2 Vincent J-L, Sakr Y, Sprung CL. et al. Sepsis in European intensive care units: Results of the SOAP study. Crit Care Med 2006; 34: 344-353.
    CrossrefPubMedGoogle Scholar
  • 3 Hoyert DL, Xu J. Deaths: Preliminary Data for 2011. National Vital Statistics Reports 2012; 61: 1-52.
    PubMedGoogle Scholar
  • 4 Kumar A, Roberts D, Wood KE. et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006; 34: 1589-1596.
    CrossrefPubMedGoogle Scholar
  • 5 Pierrakos C, Vincent J-L. Sepsis biomarkers: a review. Crit Care 2010; 14: R15.
    CrossrefPubMedGoogle Scholar
  • 6 Nordenfelt P, Winberg ME, Lönnbro P. et al. Different Requirements for Early and Late Phases of Azurophilic Granule-Phagosome Fusion. Traffic 2009; 10: 1881-1893.
    CrossrefPubMedGoogle Scholar
  • 7 Tapper H, Karlsson A, Mörgelin M. et al. Secretion of heparin-binding protein from human neutrophils is determined by its localisation in azurophilic granules and secretory vesicles. Blood 2002; 99: 1785-1793.
    CrossrefPubMedGoogle Scholar
  • 8 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685.
    CrossrefPubMedGoogle Scholar
  • 9 Sjöholm K, Karlsson C, Linder A. et al. A comprehensive analysis of the Streptococcus pyogenes and human plasma protein interaction network. Mol BioSyst. 2014 Epub ahead of print.
    PubMedGoogle Scholar
  • 10 Shevchenko A, Wilm M, Vorm O. et al. Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels. Anal Chem 1996; 68: 850-858.
    CrossrefPubMedGoogle Scholar
  • 11 Karlsson C, Malmström L, Aebersold R. et al. Proteome-wide selected reaction monitoring assays for the human pathogen Streptococcus pyogenes. Nat Commun Nature Publishing Group; 1AD; 03: 1301-1309.
    PubMedGoogle Scholar
  • 12 Malmström J, Karlsson C, Nordenfelt P. et al. Streptococcus pyogenes in Human Plasma: adaptive mechanism analysed by mass spectrometry-based proteomics. J Biol Chem 2012; 287: 1415-1425.
    CrossrefPubMedGoogle Scholar
  • 13 Craig R, Beavis RC. A method for reducing the time required to match protein sequences with tandem mass spectra. Rapid Commun Mass Spectrom 2003; 17: 2310-2316.
    CrossrefPubMedGoogle Scholar
  • 14 Keller A, Eng J, Zhang N. et al. A uniform proteomics MS/MS analysis platform utilizing open XML file formats. Mol Syst Biol 2005; 01: E1-E8.
    PubMedGoogle Scholar
  • 15 Malmström L, Malmström J, Selevsek N. et al. Automated Workflow for Large-Scale Selected Reaction Monitoring Experiments. J Proteome Res 2012; 11: 1644-1653.
    CrossrefPubMedGoogle Scholar
  • 16 Malmström L, Marko-Varga G, Westergren-Thorsson G. et al. 2DDB - a bioinformatics solution for analysis of quantitative proteomics data. BMC Bioinformatics 2006; 07: 158.
    CrossrefPubMedGoogle Scholar
  • 17 Modern Applied Statistics with S. 4 ed. Springer; New York: 2002
    Google Scholar
  • 18 Hartigan JA, Wong MA. A K-Means Clustering Algorithm. J Royal Stat Soc C 1979; 28: 100-108.
    PubMedGoogle Scholar
  • 19 Borregaard N, Cowland JB. Granules of the human neutrophilic polymorpho-nuclear leukocyte. Blood 1997; 89: 3503-3521.
    PubMedGoogle Scholar
  • 20 Martins PS, Kallas EG, Neto MC. et al. Upregulation of Reactive Oxygen Species Generation and Phagocytosis, and Increased Apoptosis in Human Neutrophils During Severe Sepsis and Septic Shock. Shock 2003; 20: 208-212.
    CrossrefPubMedGoogle Scholar
  • 21 Jog NR, Rane MJ, Lominadze G. et al. The actin cytoskeleton regulates exocytosis of all neutrophil granule subsets. AJP: Cell Physiology 2006; 292: C1690-1700.
    CrossrefPubMedGoogle Scholar
  • 22 Bainton DF, Ullyot JL, Farquhar MG. The development of neutrophilic polymorphonuclear leukocytes in human bone marrow. J Exp Med 1971; 134: 907-934.
    CrossrefPubMedGoogle Scholar
  • 23 Farrah T, Deutsch EW, Omenn GS. et al. A High-Confidence Human Plasma Proteome Reference Set with Estimated Concentrations in PeptideAtlas. Mol Cell Proteomics 2011; 10: M110.006353-3.
    CrossrefPubMedGoogle Scholar
  • 24 Linder A, Christensson B, Herwald H. et al. Heparin-Binding Protein: An Early Marker of Circulatory Failure in Sepsis. Clin Infect Dis 2009; 49: 1044-1050.
    CrossrefPubMedGoogle Scholar
  • 25 Lee SE, Kim H-S. Human resistin in cardiovascular disease. J Smooth Muscle Res 2012; 48: 27-35.
    CrossrefPubMedGoogle Scholar
  • 26 Hazlett L, Wu M. Defensins in innate immunity. Cell Tissue Res 2010; 343: 175-188.
    PubMedGoogle Scholar
  • 27 Gersemann M, Becker S, Nuding S. et al. Olfactomedin-4 is a glycoprotein secreted into mucus in active IBD. J Crohns Colitis European Crohn’s and Colitis Organisation; 2012; 06: 425-434.
    PubMedGoogle Scholar
  • 28 Tapper H, Grinstein S. Fc receptor-triggered insertion of secretory granules into the plasma membrane of human neutrophils - Selective retrieval during phagocytosis. J Immunol 1997; 159: 409-418.
    PubMedGoogle Scholar
  • 29 da Silva FP, Aloulou M, Skurnik D. et al. CD16 promotes Escherichia coli sepsis through an FcR inhibitory pathway that prevents phagocytosis and facilitates inflammation. Nat Med 2007; 13: 1368-1374.
    CrossrefPubMedGoogle Scholar
  • 30 Prokopowicz Z, Marcinkiewicz J, Katz DR. et al. Neutrophil Myeloperoxidase: Soldier and Statesman. Arch Immunol Ther Exp 2011; 60: 43-54.
    PubMedGoogle Scholar
  • 31 Son YM, Ahn SM, Jang MS. et al. Biochemical and Biophysical Research Communications. Biochem Biophys Res Commun Elsevier Inc; 2008; 376: 599-604.
    PubMedGoogle Scholar
  • 32 Moore DF, Rosenfeld MR, Gribbon PM. et al. Alpha-1-acid (AAG, orosomucoid) glycoprotein: interaction with bacterial lipopolysaccharide and protection from sepsis. Inflammation 1997; 21: 69-82.
    CrossrefPubMedGoogle Scholar
  • 33 Levy AP, Asleh R, Blum S. et al. Haptoglobin: Basic and Clinical Aspects. Anti-oxid Redox Signal 2010; 12: 293-304.
    CrossrefPubMedGoogle Scholar
  • 34 Hasan Z, Palani K, Rahman M. et al. Targeting CD44 Expressed on Neutrophils Inhibits Lung Damage in Abdominal Sepsis. Shock 2011; 35: 567-572.
    CrossrefPubMedGoogle Scholar
  • 35 Tang W, Lu Y, Tian QY. et al. The Growth Factor Progranulin Binds to TNF Receptors and Is Therapeutic Against Inflammatory Arthritis in Mice. Science 2011; 332: 478-484.
    CrossrefPubMedGoogle Scholar
  • 36 Goetz DH, Holmes MA, Borregaard N. et al. The neutrophil lipocalin NGAL is a bacteriostatic agent that interferes with siderophore-mediated iron acquisition. Mol Cell 2002; 10: 1033-1043.
    CrossrefPubMedGoogle Scholar
  • 37 Bucki R, Leszczyńska K, Namiot A. et al. Cathelicidin LL-37: A Multitask Antimicrobial Peptide. Arch Immunol Ther Exp 2010; 58: 15-25.
    CrossrefPubMedGoogle Scholar
  • 38 Xu S, Zhao L, Larsson A. et al. The identification of a phospholipase B precursor in human neutrophils. FEBS J 2008; 276: 175-86.
    PubMedGoogle Scholar
  • 39 Shi X-Z, Zhong X, Yu X-Q. Drosophila melanogaster NPC2 proteins bind bacterial cell wall components and may function in immune signal pathways. Insect Biochem Mol Biol 2012; 42: 545-556.
    CrossrefPubMedGoogle Scholar
  • 40 Lu J, Holmgren A. Free Radical Biology and Medicine. Free Radical Biology and Medicine Elsevier; 2014; 66 pp 75-87.
    PubMedGoogle Scholar
  • 41 Goyal MM, Basak A. Human catalase: looking for complete identity. Protein Cell 2010; 01: 888-897.
    CrossrefPubMedGoogle Scholar
  • 42 Whitbread AK, Masoumi A, Tetlow N. et al Characterisation of the Omega Class of Glutathione Transferases. Methods in Enzymology Elsevier. 2005 pp. 78-99.
    PubMedGoogle Scholar
  • 43 Perl A, Hanczko R, Telarico T. et al. Oxidative stress, inflammation and carcino-genesis are controlled through the pentose phosphate pathway by transaldolase. Trends Mol Med 2011; 17: 395-403.
    CrossrefPubMedGoogle Scholar
  • 44 Gabay C, Kushner I. Acute-phase proteins and other systemic responses to inflammation. N Engl J Med 1999; 340: 448-454.
    CrossrefPubMedGoogle Scholar