title: Effect of Puumala hantavirus infection on human umbilical vein endothelial cell hemostatic function: platelet interactions, increased tissue factor expression and fibrinolysis regulator release.
abstract: Puumala virus (PUUV) infection causes over 5000 cases of hemorrhagic fever in Europe annually and can influence the hemostatic balance extensively. Infection might lead to hemorrhage, while a recent study showed an increased risk of myocardial infarction during or shortly after PUUV infection. The mechanism by which this hantavirus influences the coagulation system remains unknown. Therefore we aimed to elucidate mechanisms explaining alterations seen in primary and secondary hemostasis during PUUV infection. By using low passage PUUV isolates to infect primary human umbilical vein endothelial cells (HUVECs) we were able to show alterations in the regulation of primary- and secondary hemostasis and in the release of fibrinolysis regulators. Our main finding was an activation of secondary hemostasis due to increased tissue factor (TF) expression leading to increased thrombin generation in a functional assay. Furthermore, we showed that during infection platelets adhered to HUVEC and subsequently specifically to PUUV virus particles. Infection of HUVEC with PUUV did not result in increased von Willebrand factor while they produced more plasminogen activator inhibitor type-1 (PAI-1) compared to controls. The PAI-1 produced in this model formed complexes with vitronectin. This is the first report that reveals a potential mechanism behind the pro-coagulant changes in PUUV patients, which could be the result of increased thrombin generation due to an increased TF expression on endothelial cells during infection. Furthermore, we provide insight into the contribution of endothelial cell responses regarding hemostasis in PUUV pathogenesis.
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands.
authors: Marco
authors: M
authors: Goeijenbier
authors: Department of Experimental Vascular Medicine, Academic Medical Center, University of Amsterdam Amsterdam, Netherlands ; Department of Plasma Proteins, Sanquin Research, Amsterdam Netherlands.
authors: Joost C M
authors: JC
authors: Meijers
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands ; Department of Internal Medicine, Faculty of Medicine, Universitas Indonesia Jakarta, Indonesia.
authors: Fatih
authors: F
authors: Anfasa
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands ; Artemis One Health Institute, Utrecht Netherlands.
authors: Jeroen M
authors: JM
authors: Roose
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands.
authors: Cornelia A M
authors: CA
authors: van de Weg
authors: Department of Experimental Vascular Medicine, Academic Medical Center, University of Amsterdam Amsterdam, Netherlands ; Department of Plasma Proteins, Sanquin Research, Amsterdam Netherlands.
authors: Kamran
authors: K
authors: Bakhtiari
authors: Metla, Finnish Forest Research Institute, Vantaa Finland.
authors: Heikki
authors: H
authors: Henttonen
authors: Department of Virology, Haartman Institute, University Of Helsinki Helsinki, Finland.
authors: Antti
authors: A
authors: Vaheri
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands ; Artemis One Health Institute, Utrecht Netherlands.
authors: Albert D M E
authors: AD
authors: Osterhaus
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands.
authors: Eric C M
authors: EC
authors: van Gorp
authors: Department of Viroscience, Erasmus MC, Rotterdam Netherlands ; Artemis One Health Institute, Utrecht Netherlands.
authors: Byron E E
authors: BE
authors: Martina
keywords: HFRS; endothelial cells; hantavirus; hemorrhagic fever with renal syndrome; hemostasis; platelets; thrombin generation