title: NF-κB is required for dengue virus NS5-induced RANTES expression.
abstract: Dengue virus (DENV) infection associates with renal disorders. Patients with dengue hemorrhagic fever and acute kidney injury have a high mortality rate. Increased levels of cytokines may contribute to the pathogenesis of DENV-induced kidney injury. Currently, molecular mechanisms how DENV induces kidney cell injury has not been thoroughly investigated. Excessive cytokine production may be involved in this process. Using human cytokine RT(2) Profiler PCR array, 14 genes including IP-10, RANTES, IL-8, CXCL-9 and MIP-1β were up-regulated more than 2 folds in DENV-infected HEK 293 cells compared to that of mock-infected HEK 293 cells. In the present study, RANTES was suppressed by the NF-κB inhibitor, compound A (CpdA), in DENV-infected HEK 293 cells implying the role of NF-κB in RANTES expression. Chromatin immunoprecipitation (ChIP) assay showed that NF-κB binds more efficiently to its binding sites on the RANTES promoter in NS5-transfected HEK 293 cells than in HEK 293 cells expressing the vector lacking NS5 gene. To further examine whether the NS5-activated RANTES promoter is mediated through NF-κB, the two NF-κB binding sites on the RANTES promoter were mutated and this promoter was coupled to the luciferase cDNA. The result showed that when both binding sites of NF-κB in the RANTES promoter were mutated, the ability of NS5 to induce the luciferase activity was significantly decreased. Therefore, DENV NS5 activates RANTES production by increasing NF-κB binding to its binding sites on the RANTES promoter.
authors.affiliation: Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand; Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
authors.forename: Sasiprapa
authors.initials: S
authors.lastname: Khunchai
authors.affiliation: Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
authors.forename: Mutita
authors.initials: M
authors.lastname: Junking
authors.affiliation: Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
authors.forename: Aroonroong
authors.initials: A
authors.lastname: Suttitheptumrong
authors.affiliation: Department of Physiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
authors.forename: Suwattanee
authors.initials: S
authors.lastname: Kooptiwut
authors.affiliation: Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand.
authors.forename: Guy
authors.initials: G
authors.lastname: Haegeman
authors.affiliation: Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand. Electronic address: thawornchai.lim@mahidol.ac.th.
authors.forename: Thawornchai
authors.initials: T
authors.lastname: Limjindaporn
authors.affiliation: Division of Molecular Medicine, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand. Electronic address: ptyench@gmail.com.
authors.forename: Pa-Thai
authors.initials: PT
authors.lastname: Yenchitsomanus
mesh_terms: D002460:Cell Line; D018946:Chemokine CCL5 / Q000096:biosynthesis*; D047369:Chromatin Immunoprecipitation; D003716:Dengue Virus / Q000276:immunology*; D020869:Gene Expression Profiling; D006801:Humans; D046228:Microarray Analysis; D016328:NF-kappa B / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D018946:Chemokine CCL5; D016328:NF-kappa B; C582223:NS5 protein, dengue virus; D017361:Viral Nonstructural Proteins
keywords: Dengue virus; NF-κB; NS5; RANTES