Background: Accurate identification of the proportion of young platelets is important to distinguish peripheral thrombocytopenia from a deficit in platelet production. Young platelets are defined by their higher RNA content and are often assessed as thiazole orange bright (TObright ) by flow cytometry. In clinical practice, their proportion is estimated by automatic blood counter according to their greater RNA content, which identifies a so-called immature platelet fraction (IPF). However, the detected IPFs are not strictly identical to the young TObright platelet population observed by flow cytometry.
Objectives: The aim of this study was to assess the reliability of HLA I/major histocompatibility I (MHC I) cell surface expression as a marker of young platelets.
Methods: The HLA I/MHC I expression was evaluated by flow cytometry after costaining blood with TO and antibodies directed against HLA I/MHC I molecules.
Results: We found that platelets with a higher expression of plasma membrane-localized MHC I molecules displayed an increased TO staining and a higher content in ribosomal P-antigen. Transfusion experiments in mice showed that the number of MHC I molecules expressed on the cell surface of young murine platelets decreased during platelet aging, reaching basal levels within 24 h. Finally, we demonstrated that for patients with thrombocytopenias, the identification of young platelets is better assessed by the flow cytometric determination of the level of HLA I expression than by TO staining or the use of hematological blood counter.
Conclusion: Overall, our results highlight the relevance of MHC I/HLA I expression as a valuable parameter to identify young platelets.
Keywords: HLA-I; diagnosis; flow cytometry; platelets; thrombocytopenia.
© 2019 International Society on Thrombosis and Haemostasis.