Simultaneous Measurement of Changes in Neutrophil Granulocyte Membrane Potential, Intracellular pH, and Cell Size by Multiparametric Flow Cytometry
| Autor: | Bertram Dietrich Thomaß, David A. C. Messerer, Laura Vidoni, Marco Mannes, Lisa Wohlgemuth, Stefan Bernhard, Maike Erber, Karl J. Föhr, Adam Omar Khalaf Mohamed, Michael Fauler, Markus Huber-Lang, Laura Stukan, Stefan Hug, Frederik Münnich, Alexander Elias Paul Stratmann |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Lipopolysaccharide
QH301-705.5 Neutrophil granulocyte Intracellular pH Medicine (miscellaneous) intracellular pH Context (language use) General Biochemistry Genetics and Molecular Biology Article Flow cytometry Proinflammatory cytokine chemistry.chemical_compound neutrophils medicine Biology (General) innate immunity forward scatter medicine.diagnostic_test Chemistry Multiparametric Analysis flow cytometry lipopolysaccharide Depolarization Cell biology medicine.anatomical_structure membrane potential |
| Zdroj: | Biomedicines, Vol 9, Iss 1504, p 1504 (2021) Biomedicines Volume 9 Issue 11 |
| ISSN: | 2227-9059 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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