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. 2016 Nov 1:115:14.41.1-14.41.14.
doi: 10.1002/cpim.16.

Induction and Quantification of NETosis

Carmelo Carmona-Rivera  1 , Mariana J Kaplan  1
Affiliations

Induction and Quantification of NETosis

Carmelo Carmona-Rivera et al. Curr Protoc Immunol. .

Abstract

Neutrophil extracellular traps (NET) are web-like structures composed of nuclear material and neutrophil granular proteins that are released to the extracellular space after specific microbial or sterile inflammatory stimuli. NETosis is a specific type of cell death that ensues when neutrophils extrude NETs. It is important to develop validated standards to induce and quantify NETs in order to accurately compare results between laboratories and advance the understanding of this biological process and its implications in health and disease. This unit presents methods for isolating human neutrophils from peripheral blood and inducing NETs through various types of stimulation that rely on the production of reactive oxygen species from NADPH oxidase or mitochondria. Alternative methods are also described for low- and high-throughput quantification of NETs. © 2016 by John Wiley & Sons, Inc.

Keywords: flow cytometry; immunofluorescence; neutrophil extracellular traps; neutrophils.

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References

Literature Cited

    1. Abi Abdallah, D.S. , Lin, C. , Ball, C.J. , King, M.R. , Duhamel, G. E. , and Denkers, E. Y. 2012. Toxoplasma gondii triggers release of human and mouse neutrophil extracellular traps. Infect. Immun. 80:768-777 doi: 10.1128/IAI.05730-11.
    1. Beiter, K. , Wartha, F. , Albiger, B. , Normark, S. , Zychlinsky, A. , and Henriques-Normark, B. 2006. An endonuclease allows Streptococcus pneumoniae to escape from neutrophil extracellular traps. Curr. Biol. 16:401-407. doi: 10.1016/j.cub.2006.01.056.
    1. Brinkmann, V. , Reichard, U. , Goosmann, C. , Fauler, B. , Uhlemann, Y. , Weiss, D. S. , Weinrauch, Y. , and Zychlinsky, A. 2004. Neutrophil extracellular traps kill bacteria. Science 303:1532-1535. doi: 10.1126/science.1092385.
    1. Carmona-Rivera, C. , Zhao, W. , Yalavarthi, S. , and Kaplan, M.J. 2015. Neutrophil extracellular traps induce endothelial dysfunction in systemic lupus erythematosus through the activation of matrix metalloproteinase-2. Ann. Rheum. Dis. 74:1417-1424. doi: 10.1136/annrheumdis-2013-204837. Epub 2014 Feb 25.
    1. Clark, S.R. , Ma, A.C. , Tavener, S.A. , McDonald, B. , Goodarzi, Z. , Kelly, M.M. , Patel, K.D. , Chakrabarti, S. , McAvoy, E. , Sinclair, G.D. , Keys, E.M. , Allen-Vercoe, E. , Devinney, R. , Doig, C.J , Green, F.H. , and Kubes, P. 2007. Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood. Nat. Med. 13:463-469 doi: 10.1038/nm1565.

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