Dengue virus-induced autophagosomes and changes in endomembrane ultrastructure imaged by electron tomography and whole-mount grid-cell culture techniques.
The biogenesis events and formation of dengue virus (DENV) in the infected host cells remain incompletely understood. In the present study, we examined the ultrastructural changes associated with DENV-2 replication in three susceptible host cells, C6/36, Vero and SK Hep1, a cell line of human endothelial origin, using transmission electron microscopy, whole-mount grid-cell culture techniques and electron tomography (ET). The prominent feature in C6/36 cells was the formation of large perinuclear vacuoles with mature DENV particles, and on-grid whole-mount examination of the infected Vero cells showed different forms of DENV core structures associated with cellular membranes within 48 h after infection. Distinct multivesicular structures and prominent autophagic vesicles were seen in the infected SK Hep1 cells when compared with the other two cell lines. ET showed the three-dimensional organization of these vesicles as a continuous system. This is the first report of ET-based analysis of DENV-2 replication in a human endothelial cell line. These results further emphasizes the strong role played by intracellular host membranes-virus interactions in the biogenesis of DENV and strongly argues for the possibility of targeting compounds to block such structure formation as key anti-dengue agents.
Gangodkar
Shobha
S
Electron Microscopy Group, National Institute of Virology (ICMR), 20A Dr Ambedkar Road, Pune 411001, India.
Jain
Preksha
P
Dixit
Nishikant
N
Ghosh
Kanjaksha
K
Basu
Atanu
A
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure* / Q000821:virology; D002522:Chlorocebus aethiops; D003716:Dengue Virus / Q000472:pathogenicity*; D055032:Electron Microscope Tomography / Q000379:methods; D042783:Endothelial Cells / Q000648:ultrastructure* / Q000821:virology; D006801:Humans; D046529:Microscopy, Electron, Transmission / Q000379:methods; D010588:Phagosomes / Q000648:ultrastructure*; D014709:Vero Cells; D014779:Virus Replication