title: RNA interference mediated silencing of Hsp60 gene in human monocytic myeloma cell line U937 revealed decreased dengue virus multiplication.
abstract: Heat shock proteins (Hsps) or stress proteins are highly conserved molecules and expressed in all cell types under stressful conditions like heat, cold, hypoxia and infections. The objective of the present study was to determine the effect of dengue virus infection on relative expression of stress proteins and their role in the progression of the infection. As macrophages are the primary host for dengue, human promonocytic myeloblastoma U937 cells were infected with dengue virus type 2 New Guinea C strain for the evaluation of Hsps expression. A significant expression of Hsp60 was observed in virally infected U937 cells as compared to controls. In order to determine the correlation between Hsp60 expression and viral multiplication in infected cells, expression of Hsp60 was down regulated by RNA interference. Viral multiplication was determined by quantification of viral RNA copy number using Real Time PCR and plaque formation assay in cellular supernatants of Hsp60 silenced cells. Intracellular quantification of viral load was also determined by flow cytometry. It was observed that down regulation of Hsp60 in virally infected cells resulted into decrease in viral RNA copy number, plaque forming units and intracellular viral load. At the same time down regulation also resulted in increased IFN-alpha level. These observations suggest that, elevated levels of Hsp60 expression in virally infected cells may help in viral multiplication and could be possible therapeutic targets for the management of dengue virus infection.
authors.affiliation: Immunomodulation Laboratory, Defence Institute of Physiology and Allied Sciences, Timarpur, Delhi 110054, India.
authors.forename: Y S
authors.initials: YS
authors.lastname: Padwad
authors.forename: K P
authors.initials: KP
authors.lastname: Mishra
authors.forename: M
authors.initials: M
authors.lastname: Jain
authors.forename: S
authors.initials: S
authors.lastname: Chanda
authors.forename: D
authors.initials: D
authors.lastname: Karan
authors.forename: L
authors.initials: L
authors.lastname: Ganju
mesh_terms: D018834:Chaperonin 60 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003715:Dengue / Q000235:genetics* / Q000276:immunology / Q000821:virology; D003716:Dengue Virus / Q000472:pathogenicity / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D006801:Humans; D016898:Interferon-alpha; D009000:Monocytes / Q000276:immunology / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020298:U937 Cells; D010948:Viral Plaque Assay; D014774:Virulence; D014779:Virus Replication
chemical_list: D018834:Chaperonin 60; D016898:Interferon-alpha; D034741:RNA, Small Interfering