title: Isolation and propagation of Dengue virus in Vero and BHK-21 cells expressing human DC-SIGN stably.
abstract: The "standard" methods of isolating dengue virus (DENV) utilize the mosquito cell line C6/36, monkey kidney LLC-MK2 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.
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Supranee
authors.initials: S
authors.lastname: Phanthanawiboon
authors.affiliation: National Institute of Health, Department of Medical Sciences, Muang, Nonthaburi 11000, Thailand.
authors.forename: Atchareeya
authors.initials: A
authors.lastname: A-nuegoonpipat
authors.affiliation: National Institute of Health, Department of Medical Sciences, Muang, Nonthaburi 11000, Thailand.
authors.forename: Narawan
authors.initials: N
authors.lastname: Panngarm
authors.affiliation: Department of Tropical Pediatrics, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, Thailand.
authors.forename: Kriengsak
authors.initials: K
authors.lastname: Limkittikul
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Ikuta
authors.affiliation: National Institute of Health, Department of Medical Sciences, Muang, Nonthaburi 11000, Thailand.
authors.forename: Surapee
authors.initials: S
authors.lastname: Anantapreecha
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: tkurosu@biken.osaka-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kurosu
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000235:genetics*; D002460:Cell Line; D003716:Dengue Virus / Q000254:growth & development* / Q000302:isolation & purification*; D015870:Gene Expression; D006801:Humans; D037181:Lectins, C-Type / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000235:genetics*; D011991:Receptors, Virus / Q000235:genetics*; D014776:Virus Cultivation / Q000379:methods; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D011991:Receptors, Virus; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
keywords: DC-SIGN; Dengue virus; Receptor; Virus isolation