title: Platelet adhesion to dengue-2 virus-infected endothelial cells.
abstract: Platelets in circulation normally do not adhere to resting endothelial cells. However, in response to vascular injury they adhere to stimulated endothelium and thereby play an essential role in hemostasis and thrombosis. Infection with dengue-2 virus can cause illness accompanied by thrombocytopenia and hemorrhage. Increased adherence of platelets to stimulated endothelial cells could contribute to the thrombocytopenia. In this study, adherence of radioisotopically labeled platelets to 1) unstimulated, 2) lipopolysaccharide (LPS)-stimulated, and 3) dengue-2 virus-infected human umbilical vein endothelial cells (HUVEC) was measured in an in vitro assay. Primary HUVEC were cultured in 96-well tissue culture plates in the presence or absence of LPS or dengue-2 virus. These cells were co-incubated with 3H-adenine-labeled fresh platelets for 30 min after which the cells were assayed for adherent platelets. Within 30 min there was maximum adherence of platelets to confluent LPS-stimulated HUVEC (36 +/- 4% over controls; P = 0.005). In comparison, there was a significant increase in adherence to dengue-2 infected HUVEC (78 +/- 7%; P < or = 0.001). Additionally, platelet adherence was visualized using fluorescent microscopy. Dengue-2 infection stimulated the HUVEC as monitored by expression of E-selectin. Platelets that adhered to dengue-2 or LPS-stimulated HUVEC were activated as visualized by dual fluorescent probes. These data demonstrate that human platelets adhere to dengue-2 virus-stimulated HUVEC and this interaction could contribute to the thrombocytopenia observed during infection.
authors.affiliation: Department of Blood Research, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910-7500, USA. krishnac@nhlbi.nih.gov
authors.forename: Chitra
authors.initials: C
authors.lastname: Krishnamurti
authors.forename: Raquel A
authors.initials: RA
authors.lastname: Peat
authors.forename: Mary A
authors.initials: MA
authors.lastname: Cutting
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Rothwell
mesh_terms: D001792:Blood Platelets / Q000502:physiology*; D002478:Cells, Cultured; D003716:Dengue Virus / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology / Q000821:virology*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D010973:Platelet Adhesiveness; D014471:Umbilical Veins
chemical_list: D008070:Lipopolysaccharides