Limits...
Transcriptional regulation of IFN- λ genes in Hepatitis C virus-infected hepatocytes via IRF-3 · IRF-7 · NF- κ B complex

View Article: PubMed Central - HTML

No MeSH data available.


Related in: MedlinePlus

Overexpression of IRFs and NF-κB confirms their role in expression of IFN-λ genes. PH5CH8 cells were transiently transfected with plasmids encoding different promoter regions of human IL-28A (pGL3-IL-28A 1kb) (A-C), IL-28B (pGL3-IL-28B 1kb) (D-F), or IL-29 (pGL3-IL-29 1kb) (G-J), and expression vectors encoding IRF-3, IRF-7, and NF-κB or a dominant-negative mutant of IKKβ (DNIκB). At 18 h post-transfection, cells were stimulated with poly(I:C) for 6 hours. Luciferase activity in whole cell lysates was normalized to Renilla luciferase activity. Data represent mean ± SD of triplicate data points of one experiment and are representative of five independent experiments (p<0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4288562&req=5

Figure 4: Overexpression of IRFs and NF-κB confirms their role in expression of IFN-λ genes. PH5CH8 cells were transiently transfected with plasmids encoding different promoter regions of human IL-28A (pGL3-IL-28A 1kb) (A-C), IL-28B (pGL3-IL-28B 1kb) (D-F), or IL-29 (pGL3-IL-29 1kb) (G-J), and expression vectors encoding IRF-3, IRF-7, and NF-κB or a dominant-negative mutant of IKKβ (DNIκB). At 18 h post-transfection, cells were stimulated with poly(I:C) for 6 hours. Luciferase activity in whole cell lysates was normalized to Renilla luciferase activity. Data represent mean ± SD of triplicate data points of one experiment and are representative of five independent experiments (p<0.01).

Mentions: Hepatitis C virus (HCV) infection in hepatocytes stimulates innate antiviral responses including the production of type III interferons (IFN-λ), including IL-28A, IL-28B, and IL-29 (Figure 1). However, the molecular mechanism(s) regulating the expression of IFN-λ genes in HCV-infected hepatocytes remains undefined. In this study, we examined regulatory elements involved in the induction of IFN-λ genes following HCV infection in hepatocytes and further determined the binding of specific transcription factor(s) to promoter regions of IFN-λ genes. Our studies reveal that the regulatory portion for IL-28A, IL-28B, andIL-29 genes is localized to a 1-kb region in their respective promoters (Figure 2, 4). Notably, interferon regulatory factor (IRF)-3 and -7 are the key transcriptional factors for the induction of IL-28A and IL-28Bgenes (Figure 5, 6), whereas NF-κB is an additional requirement for the induction of the IL-29 gene (Figure 3). Ligation of Toll-like receptors (TLR) 3, 7, 8, and 9, which also activate IRFs and NF-κB, resulted in more robust production of IFN-λ than that observed with HCV infection, verifying the importance of TLR pathways in IFN-λ production (Figure 8). Furthermore, the addition of IFN-λ to HCV-infected hepatocytes decreased viral replication and produced a concurrent reduction in microRNA-122 (miR-122). The decrease in viral replication was enhanced by the co-administration of IFN-λ and miR-122 inhibitor (miRIDIAN) (Figure 7), suggesting that such combinatorial therapies may be beneficial for the treatment of chronic HCV infection.


Transcriptional regulation of IFN- λ genes in Hepatitis C virus-infected hepatocytes via IRF-3 · IRF-7 · NF- κ B complex
Overexpression of IRFs and NF-κB confirms their role in expression of IFN-λ genes. PH5CH8 cells were transiently transfected with plasmids encoding different promoter regions of human IL-28A (pGL3-IL-28A 1kb) (A-C), IL-28B (pGL3-IL-28B 1kb) (D-F), or IL-29 (pGL3-IL-29 1kb) (G-J), and expression vectors encoding IRF-3, IRF-7, and NF-κB or a dominant-negative mutant of IKKβ (DNIκB). At 18 h post-transfection, cells were stimulated with poly(I:C) for 6 hours. Luciferase activity in whole cell lysates was normalized to Renilla luciferase activity. Data represent mean ± SD of triplicate data points of one experiment and are representative of five independent experiments (p<0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4288562&req=5

Figure 4: Overexpression of IRFs and NF-κB confirms their role in expression of IFN-λ genes. PH5CH8 cells were transiently transfected with plasmids encoding different promoter regions of human IL-28A (pGL3-IL-28A 1kb) (A-C), IL-28B (pGL3-IL-28B 1kb) (D-F), or IL-29 (pGL3-IL-29 1kb) (G-J), and expression vectors encoding IRF-3, IRF-7, and NF-κB or a dominant-negative mutant of IKKβ (DNIκB). At 18 h post-transfection, cells were stimulated with poly(I:C) for 6 hours. Luciferase activity in whole cell lysates was normalized to Renilla luciferase activity. Data represent mean ± SD of triplicate data points of one experiment and are representative of five independent experiments (p<0.01).
Mentions: Hepatitis C virus (HCV) infection in hepatocytes stimulates innate antiviral responses including the production of type III interferons (IFN-λ), including IL-28A, IL-28B, and IL-29 (Figure 1). However, the molecular mechanism(s) regulating the expression of IFN-λ genes in HCV-infected hepatocytes remains undefined. In this study, we examined regulatory elements involved in the induction of IFN-λ genes following HCV infection in hepatocytes and further determined the binding of specific transcription factor(s) to promoter regions of IFN-λ genes. Our studies reveal that the regulatory portion for IL-28A, IL-28B, andIL-29 genes is localized to a 1-kb region in their respective promoters (Figure 2, 4). Notably, interferon regulatory factor (IRF)-3 and -7 are the key transcriptional factors for the induction of IL-28A and IL-28Bgenes (Figure 5, 6), whereas NF-κB is an additional requirement for the induction of the IL-29 gene (Figure 3). Ligation of Toll-like receptors (TLR) 3, 7, 8, and 9, which also activate IRFs and NF-κB, resulted in more robust production of IFN-λ than that observed with HCV infection, verifying the importance of TLR pathways in IFN-λ production (Figure 8). Furthermore, the addition of IFN-λ to HCV-infected hepatocytes decreased viral replication and produced a concurrent reduction in microRNA-122 (miR-122). The decrease in viral replication was enhanced by the co-administration of IFN-λ and miR-122 inhibitor (miRIDIAN) (Figure 7), suggesting that such combinatorial therapies may be beneficial for the treatment of chronic HCV infection.

View Article: PubMed Central - HTML

No MeSH data available.


Related in: MedlinePlus