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Molecular profiling of T-helper immune genes during dengue virus infection.

Chen J, Ng MM, Chu JJ - Virol. J. (2008)

Bottom Line: Differential regulation of 41 Th genes was identified and of which 20 of those genes may contribute to immuno-pathogenesis of dengue virus infection by regulating inflammation, thrombocytopenia and vascular permeability.Among the strongly up-regulated genes were the RANTES, CC-CKR3, IRF4, CLEC2C, IL-6 and TLR6, which are potent inducer of inflammation and vascular permeability.Profiling genes obtained from this study may serve as potential biomarkers and the modulation of Th immune responses during dengue virus infection has important implications in disease outcome.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, 5 Science Drive 2, National University of Singapore, 117597 Singapore. g0800060@nus.edu.sg

ABSTRACT
In this study, we provide a comprehensive molecular profiling of the involvement of T- helper (Th) genes during dengue virus infection of different cell types. The Th gene profiles of three human cell types (monocytes, T-cells and hepatocytes) were analyzed simultaneously via array-based RT-PCR upon infection with dengue virus. Differential regulation of 41 Th genes was identified and of which 20 of those genes may contribute to immuno-pathogenesis of dengue virus infection by regulating inflammation, thrombocytopenia and vascular permeability. Among the strongly up-regulated genes were the RANTES, CC-CKR3, IRF4, CLEC2C, IL-6 and TLR6, which are potent inducer of inflammation and vascular permeability. Profiling genes obtained from this study may serve as potential biomarkers and the modulation of Th immune responses during dengue virus infection has important implications in disease outcome.

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Infectivity of human cell lines with DENV. (a) Growth curves of DENV in different human cell types. (b) Detection of DENV viral antigen (envelope protein) in different human cell types via flow cytometry analysis. All the human cells were shown to support DENV replication and high infectious virus titers were also obtained from the cells at 3 days post-infection.
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Figure 1: Infectivity of human cell lines with DENV. (a) Growth curves of DENV in different human cell types. (b) Detection of DENV viral antigen (envelope protein) in different human cell types via flow cytometry analysis. All the human cells were shown to support DENV replication and high infectious virus titers were also obtained from the cells at 3 days post-infection.

Mentions: Dengue virus is known to be able to infect different kinds of cell types in human [7]. In this study, the susceptibility of human cell lines, K562, Jurkat and HepG2 to DENV 2 virus infection was first determined. K562 is a myelogenous cell line with monocyte and granulocyte properties [8]. Jurkat is a T cell lymphoblast-like cell line [9] and HepG2 is a heptocellular liver cell line [10]. Both K562 and Jurkat cells were selected for analysis as they are of immune origin and the HepG2 cell line was selected since dengue virus is hepatotrophic. The use of human cell lines for this study has been given approval by the National University of Singapore Institutional Review Board. These human cell lines were first subjected to low passage human isolate DENV (serotype 2, Singapore strain Den2ST) infection at a multiplicity of infection of 10. At different time points post-infection, virus containing supernatant were harvested for plaque assays and the DENV-infected cells were stained for immunofluorescence detection of viral antigen (envelope protein) production via flow cytometry. All the three human cell lines are shown to be highly susceptible to dengue virus infection, producing reasonably high virus titers by 3 days post-infection (Figure 1a). Production of viral antigen was also detected in all the three cell lines by 3 days post-infection (Figure 1b).


Molecular profiling of T-helper immune genes during dengue virus infection.

Chen J, Ng MM, Chu JJ - Virol. J. (2008)

Infectivity of human cell lines with DENV. (a) Growth curves of DENV in different human cell types. (b) Detection of DENV viral antigen (envelope protein) in different human cell types via flow cytometry analysis. All the human cells were shown to support DENV replication and high infectious virus titers were also obtained from the cells at 3 days post-infection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2628356&req=5

Figure 1: Infectivity of human cell lines with DENV. (a) Growth curves of DENV in different human cell types. (b) Detection of DENV viral antigen (envelope protein) in different human cell types via flow cytometry analysis. All the human cells were shown to support DENV replication and high infectious virus titers were also obtained from the cells at 3 days post-infection.
Mentions: Dengue virus is known to be able to infect different kinds of cell types in human [7]. In this study, the susceptibility of human cell lines, K562, Jurkat and HepG2 to DENV 2 virus infection was first determined. K562 is a myelogenous cell line with monocyte and granulocyte properties [8]. Jurkat is a T cell lymphoblast-like cell line [9] and HepG2 is a heptocellular liver cell line [10]. Both K562 and Jurkat cells were selected for analysis as they are of immune origin and the HepG2 cell line was selected since dengue virus is hepatotrophic. The use of human cell lines for this study has been given approval by the National University of Singapore Institutional Review Board. These human cell lines were first subjected to low passage human isolate DENV (serotype 2, Singapore strain Den2ST) infection at a multiplicity of infection of 10. At different time points post-infection, virus containing supernatant were harvested for plaque assays and the DENV-infected cells were stained for immunofluorescence detection of viral antigen (envelope protein) production via flow cytometry. All the three human cell lines are shown to be highly susceptible to dengue virus infection, producing reasonably high virus titers by 3 days post-infection (Figure 1a). Production of viral antigen was also detected in all the three cell lines by 3 days post-infection (Figure 1b).

Bottom Line: Differential regulation of 41 Th genes was identified and of which 20 of those genes may contribute to immuno-pathogenesis of dengue virus infection by regulating inflammation, thrombocytopenia and vascular permeability.Among the strongly up-regulated genes were the RANTES, CC-CKR3, IRF4, CLEC2C, IL-6 and TLR6, which are potent inducer of inflammation and vascular permeability.Profiling genes obtained from this study may serve as potential biomarkers and the modulation of Th immune responses during dengue virus infection has important implications in disease outcome.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Yong Loo Lin School of Medicine, 5 Science Drive 2, National University of Singapore, 117597 Singapore. g0800060@nus.edu.sg

ABSTRACT
In this study, we provide a comprehensive molecular profiling of the involvement of T- helper (Th) genes during dengue virus infection of different cell types. The Th gene profiles of three human cell types (monocytes, T-cells and hepatocytes) were analyzed simultaneously via array-based RT-PCR upon infection with dengue virus. Differential regulation of 41 Th genes was identified and of which 20 of those genes may contribute to immuno-pathogenesis of dengue virus infection by regulating inflammation, thrombocytopenia and vascular permeability. Among the strongly up-regulated genes were the RANTES, CC-CKR3, IRF4, CLEC2C, IL-6 and TLR6, which are potent inducer of inflammation and vascular permeability. Profiling genes obtained from this study may serve as potential biomarkers and the modulation of Th immune responses during dengue virus infection has important implications in disease outcome.

Show MeSH
Related in: MedlinePlus