In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review.
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...
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doaj-2b7d03f564614e1cb8f3d165dff712ab2020-11-24T20:50:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01125e017647210.1371/journal.pone.0176472In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review.Tamara HoppenbrouwersAnouchska S A AutarAndi R SultanTsion E AbrahamWiggert A van CappellenAdriaan B HoutsmullerWillem J B van WamelHeleen M M van BeusekomJohan W van NeckMoniek P M de MaatMultiple 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.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.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.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.http://europepmc.org/articles/PMC5423591?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tamara Hoppenbrouwers Anouchska S A Autar Andi R Sultan Tsion E Abraham Wiggert A van Cappellen Adriaan B Houtsmuller Willem J B van Wamel Heleen M M van Beusekom Johan W van Neck Moniek P M de Maat |
spellingShingle |
Tamara Hoppenbrouwers Anouchska S A Autar Andi R Sultan Tsion E Abraham Wiggert A van Cappellen Adriaan B Houtsmuller Willem J B van Wamel Heleen M M van Beusekom Johan W van Neck Moniek P M de Maat In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review. PLoS ONE |
author_facet |
Tamara Hoppenbrouwers Anouchska S A Autar Andi R Sultan Tsion E Abraham Wiggert A van Cappellen Adriaan B Houtsmuller Willem J B van Wamel Heleen M M van Beusekom Johan W van Neck Moniek P M de Maat |
author_sort |
Tamara Hoppenbrouwers |
title |
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_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_sort |
in vitro induction of netosis: comprehensive live imaging comparison and systematic review. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2017-01-01 |
description |
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.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.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.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. |
url |
http://europepmc.org/articles/PMC5423591?pdf=render |
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