Dengue virus nonstructural protein 1 activates platelets via Toll-like receptor 4, leading to thrombocytopenia and hemorrhage
Fig 5
DENV NS1 enhances platelet aggregation ability after activation.
(A) Platelets were plated on poly-L-lysine-coated coverslips and treated with BSA or DENV NS1 (10 μg/ml) for 1 h. After fixation and permeabilization, platelets were stained with anti-CD61 polyclonal antibody and an anti-rabbit Alexa 594-conjugated antibody. (B) The light transmission of platelet-rich plasma (PRP) was measured in a Chrono-log aggregometer for 5 min after ADP (10 μM) or DENV NS1 (10 μg/ml) was added. (C) PRP was treated with BSA or DENV NS1 (10 μg/ml) (cotreated with or without 25 μg/ml 33D2 F(ab’)2, or cmIgG F(ab’)2) for 1 h and stimulated with ADP (2.5 μM). The light transmission of PRP was measured in a Chrono-log aggregometer, and (D) the maximum percentage of platelet aggregation was calculated by an AggRAM™ platelet aggregation analyzer (n = 3). **P<0.01; Kruskal-Wallis ANOVA (panel D).