生物
体外
血小板
中性粒细胞胞外陷阱
免疫学
分子生物学
细胞生物学
炎症
生物化学
作者
Maximilian Mauler,Julia Seyfert,David Haenel,Hannah Seeba,Janine Guenther,Daniela Stallmann,Claudia Schoenichen,Ingo Hilgendorf,Christoph Bode,Ingo Ahrens,Daniel Duerschmied
标识
DOI:10.1189/jlb.3ta0315-082r
摘要
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.
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