Flow cytometric assay for direct quantification of neutrophil extracellular traps in blood samples

American Journal of Hematology
Mathilde GavilletDavid A Williams

Abstract

Neutrophil extracellular traps (NETs) contribute to innate immunity as well as numerous diseases processes such as deep vein thrombosis, myocardial ischemia, and autoimmune disease. To date, most knowledge on NETs formation has been gathered via the qualitative microscopic examination of individual neutrophils in vitro, or aggregate structures in vivo. Here we describe a novel flow cytometry (FLOW)-based assay to identify and quantify NETs using antibodies against key NETs constituents, specifically DNA, modified histones, and granular enzymes. This method is applicable to both murine and human samples for the assessment of induced NETs in vitro, or detection of NETosis in vivo in blood samples. This FLOW-based method was validated by comparison with the well-established microscopy assay using two genetic mouse models previously demonstrated to show defective NETosis. It was then used on healthy human neutrophils for detection of ex vivo induced NETs and on blood samples from patients with sepsis for direct assessment of in vivo NET-forming neutrophils. This new methodology allows rapid and robust assessment of several thousand cells per sample and is independent of potential observer-bias, the two main limitations of the micro...Continue Reading

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References

Mar 29, 2003·Intensive Care Medicine·Mitchell M LevyUNKNOWN International Sepsis Definitions Conference
Apr 12, 2003·Critical Care Medicine·Mitchell M LevyUNKNOWN SCCM/ESICM/ACCP/ATS/SIS
Mar 6, 2004·Science·Volker BrinkmannArturo Zychlinsky
Jan 11, 2007·The Journal of Cell Biology·Tobias A FuchsArturo Zychlinsky
May 19, 2009·Nature Medicine·Kai KessenbrockDieter E Jenne
Aug 25, 2010·The Journal of Experimental Medicine·Pingxin LiYanming Wang
Jun 30, 2011·Journal of Leukocyte Biology·Michael Byung H LimMichael Glogauer
Jul 25, 2012·Proceedings of the National Academy of Sciences of the United States of America·Mélanie DemersDenisa D Wagner
May 8, 2013·Proceedings of the National Academy of Sciences of the United States of America·Kimberly MartinodDenisa D Wagner
Jul 19, 2013·The Journal of Clinical Investigation·Jonathan Cools-LartigueLorenzo Ferri
Sep 7, 2013·Blood·Bryan G Yipp, Paul Kubes
Dec 25, 2013·Blood·Kimberly Martinod, Denisa D Wagner

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Citations

Dec 5, 2015·European Journal of Immunology·Paul Naccache, Maria J G Fernandes
Dec 1, 2015·American Journal of Hematology·Aneta Manda-HandzlikOlga Ciepiela
Jun 14, 2016·Journal of Immunology Research·Nicole de Buhr, Maren von Köckritz-Blickwede
Jun 5, 2016·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sakiko MasudaAkihiro Ishizu
Jul 19, 2016·Joint, Bone, Spine : Revue Du Rhumatisme·Jean-Marie BerthelotMohamed Hamidou
Oct 30, 2016·Frontiers in Immunology·Giuseppe A RamirezNorma Maugeri
Nov 9, 2016·Clinica Chimica Acta; International Journal of Clinical Chemistry·Maurizio BruschiGian Marco Ghiggeri
Dec 30, 2016·The Journal of Experimental Medicine·Kimberly MartinodDenisa D Wagner
Jun 9, 2016·Journal of Dental Research·N FineM Glogauer
May 2, 2018·The Journal of Immunology : Official Journal of the American Association of Immunologists·Iliana AngelidouPanagiotis Skendros
Mar 10, 2018·International Journal of Laboratory Hematology·K H LeeF Passam
Dec 15, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Olga ZharkovaAnna-Marie Fairhurst
Apr 16, 2019·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Patrick M LelliottCevayir Coban
Jul 19, 2019·Arteriosclerosis, Thrombosis, and Vascular Biology·Charlotte ThålinHåkan Wallén
Jun 9, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Atsushi MuraoMonowar Aziz
Mar 21, 2018·Frontiers in Immunology·Ilaria J ChiccaPaul R Cooper
Feb 9, 2019·Frontiers in Immunology·Elsa NeubertLuise Erpenbeck
Jan 15, 2020·BMC Anesthesiology·Linda E CoxSimon T Schäfer
Dec 10, 2016·Frontiers in Immunology·Indira Neeli, Marko Radic
Jun 6, 2020·Frontiers in Immunology·Satarupa SenguptaVanessa Nomellini
Apr 5, 2020·The European Respiratory Journal·Howard M Kipen, Debra L Laskin
Jul 18, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Sakiko MasudaAkihiro Ishizu
Jul 6, 2017·The Journal of Pathology·Akrivi ChrysanthopoulouKonstantinos Ritis
Nov 19, 2019·Frontiers in Immunology·Naomi-Liza DenningPing Wang
Dec 7, 2019·The European Respiratory Journal·Denise J WoodingUNKNOWN Canadian Respiratory Research Network
Jul 9, 2020·Journal of Inflammation Research·Kritsanawan Sae-KhowDirekrit Chiewchengchol
Jan 27, 2017·Frontiers in Immunology·Alexandros MitsiosKonstantinos Ritis
Sep 6, 2018·Science Signaling·Akshay A D'CruzBen A Croker
Aug 26, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Paul J SmithRachel J Errington
Dec 31, 2020·Journal of Leukocyte Biology·Chuyi TanPing Wang
Dec 12, 2020·Thrombosis and Haemostasis·Chandini RangaswamyThomas Renné

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