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In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review

BACKGROUND: Multiple inducers of in vitro Neutrophil Extracellular Trap (NET) formation (NETosis) have been described. Since there is much variation in study design and results, our aim was to create a systematic review of NETosis inducers and perform a standardized in vitro study of NETosis inducer...

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Autores principales: Hoppenbrouwers, Tamara, Autar, Anouchska S. A., Sultan, Andi R., Abraham, Tsion E., van Cappellen, Wiggert A., Houtsmuller, Adriaan B., van Wamel, Willem J. B., van Beusekom, Heleen M. M., van Neck, Johan W., de Maat, Moniek P. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5423591/
https://www.ncbi.nlm.nih.gov/pubmed/28486563
http://dx.doi.org/10.1371/journal.pone.0176472
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author Hoppenbrouwers, Tamara
Autar, Anouchska S. A.
Sultan, Andi R.
Abraham, Tsion E.
van Cappellen, Wiggert A.
Houtsmuller, Adriaan B.
van Wamel, Willem J. B.
van Beusekom, Heleen M. M.
van Neck, Johan W.
de Maat, Moniek P. M.
author_facet Hoppenbrouwers, Tamara
Autar, Anouchska S. A.
Sultan, Andi R.
Abraham, Tsion E.
van Cappellen, Wiggert A.
Houtsmuller, Adriaan B.
van Wamel, Willem J. B.
van Beusekom, Heleen M. M.
van Neck, Johan W.
de Maat, Moniek P. M.
author_sort Hoppenbrouwers, Tamara
collection PubMed
description BACKGROUND: Multiple inducers of in vitro Neutrophil Extracellular Trap (NET) formation (NETosis) have been described. Since there is much variation in study design and results, our aim was to create a systematic review of NETosis inducers and perform a standardized in vitro study of NETosis inducers important in (cardiac) wound healing. METHODS: In vitro NETosis was studied by incubating neutrophils with PMA, living and dead bacteria (S. aureus and E. coli), LPS, (activated) platelets (supernatant), glucose and calcium ionophore Ionomycin using 3-hour periods of time-lapse confocal imaging. RESULTS: PMA is a consistent and potent inducer of NETosis. Ionomycin also consistently resulted in extrusion of DNA, albeit with a process that differs from the NETosis process induced by PMA. In our standardized experiments, living bacteria were also potent inducers of NETosis, but dead bacteria, LPS, (activated) platelets (supernatant) and glucose did not induce NETosis. CONCLUSION: Our systematic review confirms that there is much variation in study design and results of NETosis induction. Our experimental results confirm that under standardized conditions, PMA, living bacteria and Ionomycin all strongly induce NETosis, but real-time confocal imaging reveal different courses of events.
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spelling pubmed-54235912017-05-15 In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review Hoppenbrouwers, Tamara Autar, Anouchska S. A. Sultan, Andi R. Abraham, Tsion E. van Cappellen, Wiggert A. Houtsmuller, Adriaan B. van Wamel, Willem J. B. van Beusekom, Heleen M. M. van Neck, Johan W. de Maat, Moniek P. M. PLoS One Research Article BACKGROUND: Multiple inducers of in vitro Neutrophil Extracellular Trap (NET) formation (NETosis) have been described. Since there is much variation in study design and results, our aim was to create a systematic review of NETosis inducers and perform a standardized in vitro study of NETosis inducers important in (cardiac) wound healing. METHODS: In vitro NETosis was studied by incubating neutrophils with PMA, living and dead bacteria (S. aureus and E. coli), LPS, (activated) platelets (supernatant), glucose and calcium ionophore Ionomycin using 3-hour periods of time-lapse confocal imaging. RESULTS: PMA is a consistent and potent inducer of NETosis. Ionomycin also consistently resulted in extrusion of DNA, albeit with a process that differs from the NETosis process induced by PMA. In our standardized experiments, living bacteria were also potent inducers of NETosis, but dead bacteria, LPS, (activated) platelets (supernatant) and glucose did not induce NETosis. CONCLUSION: Our systematic review confirms that there is much variation in study design and results of NETosis induction. Our experimental results confirm that under standardized conditions, PMA, living bacteria and Ionomycin all strongly induce NETosis, but real-time confocal imaging reveal different courses of events. Public Library of Science 2017-05-09 /pmc/articles/PMC5423591/ /pubmed/28486563 http://dx.doi.org/10.1371/journal.pone.0176472 Text en © 2017 Hoppenbrouwers et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Hoppenbrouwers, Tamara
Autar, Anouchska S. A.
Sultan, Andi R.
Abraham, Tsion E.
van Cappellen, Wiggert A.
Houtsmuller, Adriaan B.
van Wamel, Willem J. B.
van Beusekom, Heleen M. M.
van Neck, Johan W.
de Maat, Moniek P. M.
In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title_full In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title_fullStr In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title_full_unstemmed In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title_short In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review
title_sort in vitro induction of netosis: comprehensive live imaging comparison and systematic review
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5423591/
https://www.ncbi.nlm.nih.gov/pubmed/28486563
http://dx.doi.org/10.1371/journal.pone.0176472
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