Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity

Neutrophils respond to a range of stimuli by releasing extracellular traps (NETs), a mesh consisting of chromatin plus granule and cytoplasmic proteins. We have investigated NET release in response to phorbol myristate acetate (PMA), <i>Pseudomonas aeruginosa</i> (PAO1), <i>Staphyl...

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Autores principales:	Heather A. Parker, Harry M. Jones, Christopher D. Kaldor, Mark B. Hampton, Christine C. Winterbourn
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	neutrophil extracellular traps NADPH oxidase inhibition myeloperoxidase hydrogen peroxide microbial stimuli of NETs Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/6705e2e852834972befe7b14e4ce83a9
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spelling	oai:doaj.org-article:6705e2e852834972befe7b14e4ce83a92021-11-25T16:28:39ZNeutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity10.3390/antiox101117912076-3921https://doaj.org/article/6705e2e852834972befe7b14e4ce83a92021-11-01T00:00:00Zhttps://www.mdpi.com/2076-3921/10/11/1791https://doaj.org/toc/2076-3921Neutrophils respond to a range of stimuli by releasing extracellular traps (NETs), a mesh consisting of chromatin plus granule and cytoplasmic proteins. We have investigated NET release in response to phorbol myristate acetate (PMA), <i>Pseudomonas aeruginosa</i> (PAO1), <i>Staphylococcus aureus</i> and <i>Candida albicans</i>, and the involvement of NADPH oxidase (NOX2) and myeloperoxidase (MPO) activities. An oxidative mechanism was involved with each stimulus, and the NOX2 inhibitor diphenylene iodonium (DPI) gave almost total inhibition. Notably, DPI added up to 60–90 min after stimulation still gave significant inhibition of subsequent NET formation. As most of the NOX2 activity had already occurred by that time, this indicates a requirement for late-stage low-level oxidant production. Inhibition of histone citrullination did not suppress NET formation, indicating that this was not the essential oxidant-dependent step. With PMA and <i>P. aeruginosa</i> PAO1, MPO activity played an important role in the induction of NETs and MPO inhibitors added up to 30–90 min after stimulation suppressed NET formation. NET formation with <i>S. aureus</i> and <i>C. albicans</i> was insensitive to MPO inhibition. Thus, MPO products are important with some stimuli but not others. Our results extend earlier observations with PMA and show that induction of NETs by microbial stimuli requires late stage oxidant production. Others have shown that NET formation involves NOX2-dependent elastase release from granules. As this is an early event, we conclude from our results that there is more than one oxidant-dependent step.Heather A. ParkerHarry M. JonesChristopher D. KaldorMark B. HamptonChristine C. WinterbournMDPI AGarticleneutrophil extracellular trapsNADPH oxidase inhibitionmyeloperoxidasehydrogen peroxidemicrobial stimuli of NETsTherapeutics. PharmacologyRM1-950ENAntioxidants, Vol 10, Iss 1791, p 1791 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	neutrophil extracellular traps NADPH oxidase inhibition myeloperoxidase hydrogen peroxide microbial stimuli of NETs Therapeutics. Pharmacology RM1-950
spellingShingle	neutrophil extracellular traps NADPH oxidase inhibition myeloperoxidase hydrogen peroxide microbial stimuli of NETs Therapeutics. Pharmacology RM1-950 Heather A. Parker Harry M. Jones Christopher D. Kaldor Mark B. Hampton Christine C. Winterbourn Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
description	Neutrophils respond to a range of stimuli by releasing extracellular traps (NETs), a mesh consisting of chromatin plus granule and cytoplasmic proteins. We have investigated NET release in response to phorbol myristate acetate (PMA), <i>Pseudomonas aeruginosa</i> (PAO1), <i>Staphylococcus aureus</i> and <i>Candida albicans</i>, and the involvement of NADPH oxidase (NOX2) and myeloperoxidase (MPO) activities. An oxidative mechanism was involved with each stimulus, and the NOX2 inhibitor diphenylene iodonium (DPI) gave almost total inhibition. Notably, DPI added up to 60–90 min after stimulation still gave significant inhibition of subsequent NET formation. As most of the NOX2 activity had already occurred by that time, this indicates a requirement for late-stage low-level oxidant production. Inhibition of histone citrullination did not suppress NET formation, indicating that this was not the essential oxidant-dependent step. With PMA and <i>P. aeruginosa</i> PAO1, MPO activity played an important role in the induction of NETs and MPO inhibitors added up to 30–90 min after stimulation suppressed NET formation. NET formation with <i>S. aureus</i> and <i>C. albicans</i> was insensitive to MPO inhibition. Thus, MPO products are important with some stimuli but not others. Our results extend earlier observations with PMA and show that induction of NETs by microbial stimuli requires late stage oxidant production. Others have shown that NET formation involves NOX2-dependent elastase release from granules. As this is an early event, we conclude from our results that there is more than one oxidant-dependent step.
format	article
author	Heather A. Parker Harry M. Jones Christopher D. Kaldor Mark B. Hampton Christine C. Winterbourn
author_facet	Heather A. Parker Harry M. Jones Christopher D. Kaldor Mark B. Hampton Christine C. Winterbourn
author_sort	Heather A. Parker
title	Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
title_short	Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
title_full	Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
title_fullStr	Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
title_full_unstemmed	Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity
title_sort	neutrophil net formation with microbial stimuli requires late stage nadph oxidase activity
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/6705e2e852834972befe7b14e4ce83a9
work_keys_str_mv	AT heatheraparker neutrophilnetformationwithmicrobialstimulirequireslatestagenadphoxidaseactivity AT harrymjones neutrophilnetformationwithmicrobialstimulirequireslatestagenadphoxidaseactivity AT christopherdkaldor neutrophilnetformationwithmicrobialstimulirequireslatestagenadphoxidaseactivity AT markbhampton neutrophilnetformationwithmicrobialstimulirequireslatestagenadphoxidaseactivity AT christinecwinterbourn neutrophilnetformationwithmicrobialstimulirequireslatestagenadphoxidaseactivity
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Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity

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