title: Platelet-neutrophil complex formation-a detailed in vitro analysis of murine and human blood samples.
abstract: Platelets form complexes with neutrophils during inflammatory processes. These aggregates migrate into affected tissues and also circulate within the organism. Several studies have evaluated platelet-neutrophil complexes as a marker of cardiovascular diseases in human and mouse. Although multiple publications have reported platelet-neutrophil complex counts, we noticed that different methods were used to analyze platelet-neutrophil complex formation, resulting in significant differences, even in baseline values. We established a protocol for platelet-neutrophil complex measurement with flow cytometry in murine and human whole blood samples. In vitro platelet-neutrophil complex formation was stimulated with ADP or PMA. We tested the effect of different sample preparation steps and cytometer settings on platelet-neutrophil complex detection and noticed false-positive counts with increasing acquisition speed. Platelet-neutrophil complex formation depends on platelet P-selectin expression, and antibody blocking of P-selectin consequently prevented ADP-induced platelet-neutrophil complex formation. These findings may help generating more comparable data among different research groups that examine platelet-neutrophil complexes as a marker for cardiovascular disease and novel therapeutic interventions.
authors.affiliation: Faculty of Biology, Heart Center, University of Freiburg, Germany and Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Mauler
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Seyfert
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Haenel
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Hannah
authors.initials: H
authors.lastname: Seeba
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Guenther
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Stallmann
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Schoenichen
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Hilgendorf
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Bode
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany ingo.ahrens@universitaets-herzzentrum.de.
authors.forename: Ingo
authors.initials: I
authors.lastname: Ahrens
authors.affiliation: Department of Cardiology and Angiology I, Heart Center, University of Freiburg, Germany daniel.duerschmied@universitaets-herzzentrum.de.
authors.forename: Daniel
authors.initials: D
authors.lastname: Duerschmied
mesh_terms: D000328:Adult; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D055815:Young Adult
chemical_list: D019007:P-Selectin; D019039:Platelet Glycoprotein GPIIb-IIIa Complex
keywords: P-selectin; flow cytometry; platelet activation