Abstract Number: PB1684
Meeting: ISTH 2020 Congress
Theme: Platelets and Megakaryocytes » Platelet Function and Interactions
Background: Platelets role as immune mediators in infection has recently been recognised. Dengue virus (DENV) infection is an increasing public health problem worldwide. NS1 is a highly conserved DENV glycoprotein associated to the infective capacity of the virus and the disease clinical outcome; recent data suggest a critical role of NS1-DENV in the thrombocytopenia present in severe dengue infection.
Aims: To study platelet response to DENV NS1 protein.
Methods: NS1-DENV was cloned into pHLSec plasmid, competent E. coli cells were transformed and plasmid DNA was extracted. HEK293 cells were transfected with plasmid DNA, liquid chromatography was used to purify the protein in its full form and in its individual domains. Platelet-rich plasma (PRP) was obtained from healthy donors and dengue patients by centrifugation of citrated blood and incubated with DENV NS1 proteins. After protein stimulation, platelets were stained with specific antibodies and cells fixed with 1% paraformaldehyde solution. Platelet activation was assessed by flow cytometry using an anti-CD41-PC7 antibody. Expression of platelet activation markers P-selectin and GpIIb-IIIa was assessed using anti-CD62P-PE and PAC1-FITC antibodies, respectively. The activation patterns obtained were compared with those obtained with classical platelet agonists ADP, collagen and epinephrine. Platelet aggregation was also assessed using NS1 DENV full protein and its individual domains as agonists.
Results: Flow cytometry analysis showed that the NS1 full protein as well as NS1 wing domain are able to induce the expression of P-selectin receptor and GpIIb-IIIa complex on platelet surface. DENV NS1 Full but not its domains induce platelet aggregation. Dengue patients show increased platelet activation and aggregation patterns.
Conclusions: DENV Protein NS1 induces platelet activation and aggregation. These results may partially explain the cause of thrombocytopenia in DENV virus infection and emphasize both the critical role of DV NS1 in dengue pathogenesis and the important role of platelets in host defense against viruses.
To cite this abstract in AMA style:
Viveros-Sandoval ME, Garcia-Larragoiti NI, Kim Y-, Lopez-Castaneda SE, Cano-Mendez A, Reyes-Sandoval A. Platelet Activation and Aggregation Induced by NS1 Dengue Protein [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/platelet-activation-and-aggregation-induced-by-ns1-dengue-protein/. Accessed April 17, 2024.« Back to ISTH 2020 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/platelet-activation-and-aggregation-induced-by-ns1-dengue-protein/