Using six-colour flow cytometry to analyse the activation and interaction of platelets and leukocytes – A new assay suitable for bench and bedside conditions

摘要: Abstract Introduction Platelets are main effector cells in haemostasis and also promote inflammation. Platelet-leukocyte complexes key mediators a variety of thromboinflammatory disorders consecutive organ failure. Cell-specific epitopes activation markers on platelets leukocytes can be measured using flow cytometry. However, until recently major restriction has been paucity antibody combinations lack detection strategies. We aimed to develop six-colour cytometry method which depicts multiple aspects platelet leukocyte interactions human whole blood. Materials Methods Platelets, including microparticles aggregates, were detected platelet-specific anti-CD41-FITC size-defined regions. The morphology platelet-leukocyte (including granulocyte monocyte content) depicted anti-CD45-PerCP, anti-CD66b-PE-Cy7, anti-CD14-APC antibodies single sample. Expression P-selectin CD11b anti-CD62P-PE anti-CD11b-BV421 antibodies, respectively. Results sensitivity this assay detect the effects various agonists (TRAP-6, ADP, collagen, epinephrine, TNF-α LPS) is demonstrated. Furthermore, shown induced by extracorporeal circulation vitro. suitability for bedside analysis demonstrated exemplarily patient treated with mechanical circulatory life support. Conclusions Using concurrent assessment parameters, gives detailed insights into complexity dynamics interactions. This carries potential increase our understanding mechanisms pathophysiology interaction research laboratory clinical setting.