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The effect of hydroxychloroquine on platelet activation in model experiments

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Abstract

Hydroxychloroquine (HCQ) is an antimalarial agent with pleiotropic effects and now represents a cornerstone in the management of patients with autoimmune conditions. While clinical series suggest anti-thrombotic properties, the way in which HCQ exerts this effect remains to be fully explained. Following a 24-h incubation of human umbilical vein endothelial cells (HUVEC) and human umbilical arterial endothelial cells (HUAEC) with HCQ (concentration 500, 1000 and 2000 ng/ml), these cells were then stimulated for an hour with tumor necrosis factor alpha (TNF-α) and were subsequently incubated in direct contact with thrombin-activated platelets. The expression of CD40L on platelets was measured by flow cytometry. The expression of CD40L on platelets significantly increased after direct incubation with 1000 ng/ml and 2000 ng/ml concentrations of HCQ. In contrast, after pre-incubation of HUAECs with 1000 ng/ml HCQ and following stimulation with platelets the expression of CD40L was significantly reduced also after stimulation with thrombin and TNF-α activated platelets. It was shown that the expression of CD40L on the platelets was not significantly reduced by different HCQ concentrations after contact with HCQ pre-incubated HUVECs. HCQ reduces the stimulatory effect of thrombin and TNF-α on platelet activation in the presence of endothelial cells. Our experiments suggest that HCQ pre-incubated HUAEC cells result in a reduced platelets activation measured by means of CD40L expression. Further, our results show that direct HCQ incubation of platelets (without the presence of EC) increased the expression of CD40L suggesting that the observed effect of HCQ on platelet activation may be EC mediated.

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Acknowledgements

The data has never been presented in the scientific meeting. All authors are involved in conception, design, acquisition of data, analysis, interpretation of data and revising it critically for important intellectual content. Nobody else was involved who do not meet the criteria of authorship. Authors have the permission to acknowledge from all those mentioned in the above section.

Funding

All sources of funding are declared in the Acknowledgements. All authors certify that all financial and material support for the conduct of this study clearly described in the Acknowledgements section of the manuscript. This research received no grant from any funding agency in the public, commercial or not-for-profit sectors, only for cost of supplies.

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Author notes

  1. Karen Schreiber and Savino Sciascia have contributed equally to this work.

Authors and Affiliations

  1. Danish Hospital for Rheumatic Diseases, Sønderborg, Denmark

    Karen Schreiber

  2. Danish Institute of Regional Health Research, University of Southern Denmark, Odense, Denmark

    Karen Schreiber

  3. Thrombosis and Haemostasis, Guy’s and St Thomas’ NHS Foundation Trust, London, UK

    Karen Schreiber

  4. Center of Research of Immunopathology and Rare Diseases- Coordinating Center of Piemonte and Valle D’Aosta Network for Rare Diseases, Department of Clinical and Biological Sciences, and SCDU Nephrology and Dialysis, S. Giovanni Bosco Hospital and University of Turin, Turin, Italy

    Savino Sciascia

  5. Fifth Department of Medicine (Nephrology/Endocrinology/Rheumatology), University Medical Centre Mannheim, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

    Florian Wehrmann, Jan Leipe, Bernhard K. Krämer & Ksenija Stach

  6. Department of Medical Statistics, University Medical Centre Mannheim, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany

    Christel Weiß

  7. European Center for Angioscience, University Medical Centre Mannheim, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany

    Jan Leipe, Bernhard K. Krämer & Ksenija Stach

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  1. Karen Schreiber
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  2. Savino Sciascia
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Contributions

Design: KS, KS, Data collection: FW, Data analysis: SS, Statistics: CW, Critical revision of article: JL, BKK.

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Correspondence to Ksenija Stach.

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Schreiber, K., Sciascia, S., Wehrmann, F. et al. The effect of hydroxychloroquine on platelet activation in model experiments. J Thromb Thrombolysis 52, 674–679 (2021). https://doi.org/10.1007/s11239-020-02325-y

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  • DOI: https://doi.org/10.1007/s11239-020-02325-y

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