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

Biomedicines

Karl Josef Föhr¹  Stefan Hug²  Bertram Dietrich Thomaß²  Alexander Elias Paul Stratmann²  Frederik Münnich²  Marco Mannes²  Stefan Bernhard²  Laura Stukan²  David Alexander Christian Messerer²  Markus Stefan Huber-Lang²  Lisa Wohlgemuth²  Maike Elisabeth Erber²  Laura Vidoni²  Adam Omar Khalaf Mohamed²  Michael Fauler³ 

[1] Department of Anesthesiology and Intensive Care Medicine, University Hospital Ulm, 89081 Ulm, Germany;Institute of Clinical and Experimental Trauma Immunology, University Hospital Ulm, 89081 Ulm, Germany;Institute of General Physiology, Ulm University, 89081 Ulm, Germany;

关键词: neutrophils;    innate immunity;    membrane potential;    forward scatter;    intracellular pH;    flow cytometry;   

DOI  :  10.3390/biomedicines9111504

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown