Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry

Neutrophils provide rapid and efficient defense mechanisms against invading pathogens. Upon stimulation with proinflammatory mediators, including complement factors and bacterial peptides, neutrophils respond with changes in their membrane potential, intracellular pH, and cellular size. This study p...

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Autores principales:	Alexander Elias Paul Stratmann, Lisa Wohlgemuth, Maike Elisabeth Erber, Stefan Bernhard, Stefan Hug, Michael Fauler, Laura Vidoni, Adam Omar Khalaf Mohamed, Bertram Dietrich Thomaß, Frederik Münnich, Laura Stukan, Karl Josef Föhr, Marco Mannes, Markus Stefan Huber-Lang, David Alexander Christian Messerer
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	neutrophils innate immunity membrane potential forward scatter intracellular pH flow cytometry Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/d686275d19eb4ea392ad8bb551c7eabe
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spelling	oai:doaj.org-article:d686275d19eb4ea392ad8bb551c7eabe2021-11-25T16:48:20ZSimultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry10.3390/biomedicines91115042227-9059https://doaj.org/article/d686275d19eb4ea392ad8bb551c7eabe2021-10-01T00:00:00Zhttps://www.mdpi.com/2227-9059/9/11/1504https://doaj.org/toc/2227-9059Neutrophils provide rapid and efficient defense mechanisms against invading pathogens. Upon stimulation with proinflammatory mediators, including complement factors and bacterial peptides, neutrophils respond with changes in their membrane potential, intracellular pH, and cellular size. This study provides an approach to quantify these important changes simultaneously using multiparametric flow cytometry, thereby revealing a typical sequence of neutrophil activation consisting of depolarization, alkalization, and increase in cellular size. Additionally, the time resolution of the flow cytometric measurement is improved in order to allow changes that occur within seconds to be monitored, and thus to enhance the kinetic analysis of the neutrophil response. The method is appropriate for the reliable semiquantitative detection of small variations with respect to an increase, no change, and decrease in those parameters as demonstrated by the screening of various proinflammatory mediators. As a translational outlook, the findings are put into context in inflammatory conditions in vitro as well as in a clinically relevant whole blood model of endotoxemia. Taken together, the multiparametric analysis of neutrophil responsiveness regarding depolarization, alkalization, and changes in cellular size may contribute to a better understanding of neutrophils in health and disease, thus potentially yielding innovative mechanistic insights and possible novel diagnostic and/or prognostic approaches.Alexander Elias Paul StratmannLisa WohlgemuthMaike Elisabeth ErberStefan BernhardStefan HugMichael FaulerLaura VidoniAdam Omar Khalaf MohamedBertram Dietrich ThomaßFrederik MünnichLaura StukanKarl Josef FöhrMarco MannesMarkus Stefan Huber-LangDavid Alexander Christian MessererMDPI AGarticleneutrophilsinnate immunitymembrane potentialforward scatterintracellular pHflow cytometryBiology (General)QH301-705.5ENBiomedicines, Vol 9, Iss 1504, p 1504 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	neutrophils innate immunity membrane potential forward scatter intracellular pH flow cytometry Biology (General) QH301-705.5
spellingShingle	neutrophils innate immunity membrane potential forward scatter intracellular pH flow cytometry Biology (General) QH301-705.5 Alexander Elias Paul Stratmann Lisa Wohlgemuth Maike Elisabeth Erber Stefan Bernhard Stefan Hug Michael Fauler Laura Vidoni Adam Omar Khalaf Mohamed Bertram Dietrich Thomaß Frederik Münnich Laura Stukan Karl Josef Föhr Marco Mannes Markus Stefan Huber-Lang David Alexander Christian Messerer Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
description	Neutrophils provide rapid and efficient defense mechanisms against invading pathogens. Upon stimulation with proinflammatory mediators, including complement factors and bacterial peptides, neutrophils respond with changes in their membrane potential, intracellular pH, and cellular size. This study provides an approach to quantify these important changes simultaneously using multiparametric flow cytometry, thereby revealing a typical sequence of neutrophil activation consisting of depolarization, alkalization, and increase in cellular size. Additionally, the time resolution of the flow cytometric measurement is improved in order to allow changes that occur within seconds to be monitored, and thus to enhance the kinetic analysis of the neutrophil response. The method is appropriate for the reliable semiquantitative detection of small variations with respect to an increase, no change, and decrease in those parameters as demonstrated by the screening of various proinflammatory mediators. As a translational outlook, the findings are put into context in inflammatory conditions in vitro as well as in a clinically relevant whole blood model of endotoxemia. Taken together, the multiparametric analysis of neutrophil responsiveness regarding depolarization, alkalization, and changes in cellular size may contribute to a better understanding of neutrophils in health and disease, thus potentially yielding innovative mechanistic insights and possible novel diagnostic and/or prognostic approaches.
format	article
author	Alexander Elias Paul Stratmann Lisa Wohlgemuth Maike Elisabeth Erber Stefan Bernhard Stefan Hug Michael Fauler Laura Vidoni Adam Omar Khalaf Mohamed Bertram Dietrich Thomaß Frederik Münnich Laura Stukan Karl Josef Föhr Marco Mannes Markus Stefan Huber-Lang David Alexander Christian Messerer
author_facet	Alexander Elias Paul Stratmann Lisa Wohlgemuth Maike Elisabeth Erber Stefan Bernhard Stefan Hug Michael Fauler Laura Vidoni Adam Omar Khalaf Mohamed Bertram Dietrich Thomaß Frederik Münnich Laura Stukan Karl Josef Föhr Marco Mannes Markus Stefan Huber-Lang David Alexander Christian Messerer
author_sort	Alexander Elias Paul Stratmann
title	Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
title_short	Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
title_full	Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
title_fullStr	Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
title_full_unstemmed	Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
title_sort	simultaneous measurement of changes in neutrophil granulocyte membrane potential, intracellular ph, and cell size by multiparametric flow cytometry
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/d686275d19eb4ea392ad8bb551c7eabe
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Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry

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