Publication: Isolation and propagation of Dengue virus in Vero and BHK-21 cells expressing human DC-SIGN stably
Issued Date
2014-12-01
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ISSN
18790984
01660934
01660934
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2-s2.0-84924965637
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Virological Methods. Vol.209, (2014), 55-61
Suggested Citation
Supranee Phanthanawiboon, Atchareeya A-nuegoonpipat, Narawan Panngarm, Kriengsak Limkittikul, Kazuyoshi Ikuta, Surapee Anantapreecha, Takeshi Kurosu Isolation and propagation of Dengue virus in Vero and BHK-21 cells expressing human DC-SIGN stably. Journal of Virological Methods. Vol.209, (2014), 55-61. doi:10.1016/j.jviromet.2014.08.023 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/33944
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Title
Isolation and propagation of Dengue virus in Vero and BHK-21 cells expressing human DC-SIGN stably
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Abstract
© 2014. The "standard" methods of isolating dengue virus (DENV) utilize the mosquito cell line C6/36, monkey kidney LLC-MK<inf>2</inf> cells, Vero cells, or baby hamster kidney (BHK-21) cells. However, these cells lines lack a particular DENV receptor, known as dendritic cell-specific ICAM-3-grabbing non-integrin (DC-SIGN), which is expressed on immature dendritic cells and monocytes/macrophages. This may result in less efficient virus isolation and propagation. The present study used a lentivirus vector to establish Vero and BHK-21 cell lines (Vero-DC and BHK-DC) that express human DC-SIGN stably. Five DENV strains, each passaged several times in C6/36 cells, replicated more efficiently in Vero-DC and BHK-DC than in the parental Vero or BHK-21 cells. Vero/Vero-DC and BHK-21/BHK-DC were used to isolate virus from buffy coats and plasma samples derived from 13 patients infected with DENV. Most of the viruses showed increased production in cell lines expressing DC-SIGN. However, the isolation rate was lower (15.4-46.2%) than that from C6/36 cells (84.6%). Interestingly, when the viruses were isolated in C6/36 cells prior to infecting Vero/Vero-DC and BHK-21/BHK-DC, the rate of virus production increased markedly, reaching levels higher than those initially achieved in C6/36 cells. These data suggest that Vero-DC and BHK-DC could be useful tools for virus propagation, and that human specimens may contain a factor that interferes with virus growth in mammalian cells.