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Journal of Electron Microscopy Advance Access published online on May 8, 2008
Journal of Electron Microscopy, doi:10.1093/jmicro/dfn007
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Imaging the interaction between dengue 2 virus and human blood platelets using atomic force and electron microscopy

Kanjaksha Ghosh2, Shobha Gangodkar1, Preksha Jain1, Shrimati Shetty2, Sandhya Ramjee1, Pankaj Poddar3 and Atanu Basu1,*

1 National Institute of Virology (ICMR), 20A Dr Ambedkar Road, Pune 411001
2 National Institute of Immunohematology, 13th Floor KEM Hospital, Parel, Mumbai
3 National Chemical Laboratory, Pune, India

* To whom correspondence should be addressed. E-mail: basua{at}icmr.org.in

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 104 LD50 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.

Keywords     AFM, electron microscopy, dengue virus, platelet

Received      5 October 2007, accepted 2 April 2008


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