Standardization of a Flow Cytometry Platelet Sialylation Assay, and reference Values for Beta-Galactose Exposure in Healthy Subjects

D. Lasne^1,2, T. Pascreau^1,2, S. Darame^1,2, M.-C. Bourrienne¹, P. Tournoux¹, A. Philippe¹, S. Ziachahabi¹, F. Suarez³, A. Marcais³, A. Dupont^4,5, C. Denis², A. Kauskot², D. Borgel^1,2

¹AP-HP, Hôpital Necker, Department of Biological Hematology, Paris, France, ²INSERM U 1176, Le Kremlin-Bicêtre, France, ³AP-HP, Hôpital Necker, Paris, France, ⁴CHU de Lille, Department of Hematology, Lille, France, ⁵Université de Lille, UMR INSERM 1011, Institut Pasteur de Lille, Lille, France

Abstract Number: PB0441

Meeting: ISTH 2020 Congress

Theme: Diagnostics and OMICs » Biomarkers of Thrombosis and Hemostasis

Background: A correct identification of the causes of thrombocytopenia is crucial for the appropriate management of the patients. The major mechanisms underlying a reduced platelet count are decreased production and/or increased destruction of platelets. It is now known that, in many cases, multiple mechanisms may contribute to the development of thrombocytopenia, and hyposialylation/desialylation characterized by an increase of β-galactose exposure accelerate platelet clearance.

Aims: A reference range of β-galactose exposure in the healthy population was never defined. Therefore, this step is a prerequisite before to identify thrombocytopenic diseases linked to sialylation variation, paving the way for a more widespread use of oseltamivir treatment in the future.

Methods: Our goal in this study was to develop a robust and standardized flow cytometry test in platelet-rich plasma by using RCA lectin allowing β-galactose exposure measurement in a clinical lab.

Results: Several pre-analytical and analytical conditions have been evaluated (RCA concentration, stability, platelet count and the storage conditions after blood sampling, positive and negative controls). We proposed that platelet β-galactose exposure has to be expressed with a ratio R^RCA/FSC taking into account platelet size. We then used this test to study platelet β-galactose exposure variability in a healthy population (127 adults) and we defined normal ranges. The analytical performances of our assay are good with acceptable precision results (within-run and between-day CV less than 12%).

Conclusions: Having reliable reference values is crucial for obtaining an adequate diagnostic accuracy. Based on our results, further investigations in large group of patients with thrombocytopenia will determine the value of this new biomarker.

To cite this abstract in AMA style:

Lasne D, Pascreau T, Darame S, Bourrienne M-, Tournoux P, Philippe A, Ziachahabi S, Suarez F, Marcais A, Dupont A, Denis C, Kauskot A, Borgel D. Standardization of a Flow Cytometry Platelet Sialylation Assay, and reference Values for Beta-Galactose Exposure in Healthy Subjects [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/standardization-of-a-flow-cytometry-platelet-sialylation-assay-and-reference-values-for-beta-galactose-exposure-in-healthy-subjects/. Accessed September 29, 2023.

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