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DOI: 10.1055/s-0038-1675574
Functional Classification of Paediatric Patients with Non-syndromic Delta-Storage Pool Deficiency
Publication History
22 March 2018
28 June 2018
Publication Date:
21 November 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00034925/201904/lookinside/thumbnails/10-1055-s-0038-1675574_180013-1.jpg)
Abstract
Storage pool disease (SPD) covers a group of platelet defects in which α- and/or delta-granules are reduced or cannot be secreted adequately in response to agonists. The detection of delta-granule release defects is hampered by a lack of fast and feasible tests. We aimed to implement a flow cytometry–based kinetic mepacrine assay to better identify and subgroup childhood patients with a mild to moderate bleeding diathesis and compare our method to established laboratory tests. We analysed 50 children with suspected SPD whose initial parameters were re-assessed in a second site visit. Mepacrine uptake and release patterns were correlated with CD63 exposure, platelet ADP/ATP release and content, and the bleeding score ascertained by the ISTH-BAT. Mepacrine release was overall significantly reduced in investigated patients compared with controls. Summarizing, our time-resolved approach proved to be a quick and inexpensive tool that was additionally able to distinguish between mepacrine uptake, mepacrine release, and combined defects. Classification of patients using such a kinetic assay makes it feasible to sensitively detect frequently missed SPD and to group these patients for further analyses and clinical correlations.
Zusammenfassung
Storage pool disease (SPD) umfasst eine Gruppe von Thrombozytendefekten, bei denen die α und/oder dichten Granula vermindert vorliegen bzw. nach Stimulation mit Agonisten nicht ausreichend ausgeschüttet werden können. Die Detektion von Freisetzungsdefekten der dichten Granula ist erschwert durch das Fehlen schneller und verlässlicher Tests. In dieser Arbeit haben wir einen durchflusszytometrischen kinetischen Mepacrin-Assay etabliert, um kindliche Patienten mit milder bis moderater Blutungsneigung besser identifizieren und gruppieren zu können und unseren Test mit anderen gängigen Methoden zu vergleichen. Wir haben dazu 50 Kinder mit Verdacht auf SPD analysiert, welche auch bei wiederholter Einbestellung ähnliche Blutungsparameter zeigten. Die Mepacrinaufnahme und –freisetzungsmuster wurden mit der Oberflächenexpression von CD63 auf Thrombozyten, der ADP/ATP-Freisetzung und dem ADP-Gehalt sowie dem Blutungsscore, nach den Kriterien der ISTH-BAT, korreliert. Die Mepacrinfreisetzung war insgesamt in der untersuchten Patientenkohorte signifikant reduziert verglichen zu Gesundspenderkontrollen. Zusammenfassend stellt sich unser zeitaufgelöster Ansatz als schnelles und kostengünstiges Hilfsmittel heraus, welches es möglich macht, zwischen Aufnahme-, Freisetzungs- und kombinierten Defekten zu unterscheiden. Die Klassifizierung von Patienten nach Anwendung dieser kinetischen Methode erlaubt es, häufig unerkannte SPD sensitiv zu detektieren und Patienten für weiterführende Untersuchungen und klinische Korrelationen zu gruppieren.
Authors' Contributions
H.S. and O.A. planned the study and obtained approval of the local institutional review board; S.E., J.E., and O.A. enrolled and treated patients; C.S. performed whole blood aggregometry and luminometry; J.E. and G.M. analysed blood samples by flow cytometry; G.M. and H.S. wrote the manuscript.
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