title: Imaging the interaction between dengue 2 virus and human blood platelets using atomic force and electron microscopy.
abstract: Thrombocytopenia is frequently associated with dengue virus infection. Host factors such as anti-platelet immunopathogenic processes have been implicated in the origin of dengue-associated thrombocytopenia but the role of dengue virus in directly interacting with platelets and altering their hemostatic property remains incompletely understood. In the present study, we examined the effect of dengue 2 virus on the morphology and physiological activation profile of normal human platelets using atomic force microscopy, electron microscopy and flowcytometry. Platelets obtained from healthy donors were exposed to a cell culture-adapted 10(4) LD(50) dengue 2 virus isolate in vitro and the subsequent effect on morphology and activation biology studied. Our results show that dengue 2 virus exposure at doses comparable to natural viremic states in human infections can activate platelets with an increase in P-selectin expression and fibrinogen-binding property. Atomic force, scanning and transmission electron microscopy also showed typical activation-related morphological changes such as altered platelet membrane architecture, degranulation, presence of filopodia and dilatation of the open canalicular system in the dengue 2 virus-exposed platelets but not in the controls. Importantly, Japanese encephalitis virus exposure at the same dose did not activate platelets or show any morphological changes. Our findings suggest that dengue 2 virus may directly interact with and activate platelets - an event that might be important in the origin of dengue-associated thrombocytopenia. Detailed molecular characterization of this effect might provide key knowledge toward better prophylaxis of the hemostatic complications of dengue disease.
authors.affiliation: National Institute of Virology (ICMR), 20A Dr Ambedkar Road, Pune 411001, India.
authors.forename: Kanjaksha
authors.initials: K
authors.lastname: Ghosh
authors.forename: Shobha
authors.initials: S
authors.lastname: Gangodkar
authors.forename: Preksha
authors.initials: P
authors.lastname: Jain
authors.forename: Shrimati
authors.initials: S
authors.lastname: Shetty
authors.forename: Sandhya
authors.initials: S
authors.lastname: Ramjee
authors.forename: Pankaj
authors.initials: P
authors.lastname: Poddar
authors.forename: Atanu
authors.initials: A
authors.lastname: Basu
mesh_terms: D001792:Blood Platelets / Q000502:physiology* / Q000648:ultrastructure; D003716:Dengue Virus / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D010973:Platelet Adhesiveness