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Respiratory syncytial virus (RSV) attachment and nonstructural proteins modify the type I interferon response associated with suppressor of cytokine signaling (SOCS) proteins and IFN-stimulated gene-15 (ISG15).

Moore EC, Barber J, Tripp RA - Virol. J. (2008)

Bottom Line: Respiratory syncytial virus (RSV) is a major cause of severe lower airway disease in infants and young children, but no safe and effective RSV vaccine is yet available.In the present study, we investigate suppressor of cytokine signaling (SOCS)-1 and SOCS3 expression associated with the type I IFN and IFN-stimulated gene (ISG)-15 response following infection of mouse lung epithelial (MLE-15) cells with RSV or RSV mutant viruses lacking the G gene, or NS1 and NS2 gene deletions.These results show an important role for SOCS1 regulation of the antiviral host response to RSV infection, and demonstrate a novel role for RSV G protein manipulation of SOCS3 and modulation of ISG15 and IFNbeta mRNA expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Diseases, Center for Disease Intervention, University of Georgia, Athens, GA 30602, USA. ecmoore@uga.edu

ABSTRACT
Respiratory syncytial virus (RSV) is a major cause of severe lower airway disease in infants and young children, but no safe and effective RSV vaccine is yet available. Factors attributing to this problem are associated with an incomplete understanding of the mechanisms by which RSV modulates the host cell response to infection. In the present study, we investigate suppressor of cytokine signaling (SOCS)-1 and SOCS3 expression associated with the type I IFN and IFN-stimulated gene (ISG)-15 response following infection of mouse lung epithelial (MLE-15) cells with RSV or RSV mutant viruses lacking the G gene, or NS1 and NS2 gene deletions. Studies in MLE-15 cells are important as this cell line represents the distal bronchiolar and alveolar epithelium of mice, the most common animal model used to evaluate the host cell response to RSV infection, and exhibit morphologic characteristics of alveolar type II cells, a primary cell type targeted during RSV infection. These results show an important role for SOCS1 regulation of the antiviral host response to RSV infection, and demonstrate a novel role for RSV G protein manipulation of SOCS3 and modulation of ISG15 and IFNbeta mRNA expression.

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ISG15 expression is increased in the absence of G protein expression. MLE-15 cells were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. ISG15 message expression was measured by real-time PCR (A). Transcript levels were normalized to hypoxanthine guanine phosphoribosyl transferase (HPRT) expression and calibrated to the mock condition. (B) RSV stimulation of ISG15 protein expression was determined in MLE-15 cells that were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. Cells were harvested and ISG15 levels determined by flow cytometry. Data is presented as fold-differences in protein expression relative to mock-infected cells. Differences in fold expression between virus infection groups were evaluated by Mann-Whitney U test and noted as significant as denoted by an asterisk. Data are shown as means ± standard errors (SE) of the means.
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Figure 4: ISG15 expression is increased in the absence of G protein expression. MLE-15 cells were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. ISG15 message expression was measured by real-time PCR (A). Transcript levels were normalized to hypoxanthine guanine phosphoribosyl transferase (HPRT) expression and calibrated to the mock condition. (B) RSV stimulation of ISG15 protein expression was determined in MLE-15 cells that were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. Cells were harvested and ISG15 levels determined by flow cytometry. Data is presented as fold-differences in protein expression relative to mock-infected cells. Differences in fold expression between virus infection groups were evaluated by Mann-Whitney U test and noted as significant as denoted by an asterisk. Data are shown as means ± standard errors (SE) of the means.

Mentions: Expression of the interferon-stimulated gene, ISG15, was determined in RSV and RSV mutant virus infected MLE-15 cells (Figure 4). ISG15 has been shown to modify several important molecules linked to and affecting type I interferon signal transduction, is released from cells to mediate extracellular cytokine-like activities, and evidence suggests that IFNβ and ISG15 are induced in parallel as a primary response to infection [1,38,41,42]. The level of ISG15 mRNA expression (Figure 4A) was similar to the level of ISG15 protein expression at 24 h and 48 h pi where similar levels were observed following WT or ΔNS1/2 infection of MLE-15 cells.


Respiratory syncytial virus (RSV) attachment and nonstructural proteins modify the type I interferon response associated with suppressor of cytokine signaling (SOCS) proteins and IFN-stimulated gene-15 (ISG15).

Moore EC, Barber J, Tripp RA - Virol. J. (2008)

ISG15 expression is increased in the absence of G protein expression. MLE-15 cells were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. ISG15 message expression was measured by real-time PCR (A). Transcript levels were normalized to hypoxanthine guanine phosphoribosyl transferase (HPRT) expression and calibrated to the mock condition. (B) RSV stimulation of ISG15 protein expression was determined in MLE-15 cells that were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. Cells were harvested and ISG15 levels determined by flow cytometry. Data is presented as fold-differences in protein expression relative to mock-infected cells. Differences in fold expression between virus infection groups were evaluated by Mann-Whitney U test and noted as significant as denoted by an asterisk. Data are shown as means ± standard errors (SE) of the means.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 4: ISG15 expression is increased in the absence of G protein expression. MLE-15 cells were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. ISG15 message expression was measured by real-time PCR (A). Transcript levels were normalized to hypoxanthine guanine phosphoribosyl transferase (HPRT) expression and calibrated to the mock condition. (B) RSV stimulation of ISG15 protein expression was determined in MLE-15 cells that were mock-infected or infected with WT, ΔG, or ΔNS1/2 virus at a multiplicity of infection (MOI) of 1 for 24 h or 48 h as indicated. Cells were harvested and ISG15 levels determined by flow cytometry. Data is presented as fold-differences in protein expression relative to mock-infected cells. Differences in fold expression between virus infection groups were evaluated by Mann-Whitney U test and noted as significant as denoted by an asterisk. Data are shown as means ± standard errors (SE) of the means.
Mentions: Expression of the interferon-stimulated gene, ISG15, was determined in RSV and RSV mutant virus infected MLE-15 cells (Figure 4). ISG15 has been shown to modify several important molecules linked to and affecting type I interferon signal transduction, is released from cells to mediate extracellular cytokine-like activities, and evidence suggests that IFNβ and ISG15 are induced in parallel as a primary response to infection [1,38,41,42]. The level of ISG15 mRNA expression (Figure 4A) was similar to the level of ISG15 protein expression at 24 h and 48 h pi where similar levels were observed following WT or ΔNS1/2 infection of MLE-15 cells.

Bottom Line: Respiratory syncytial virus (RSV) is a major cause of severe lower airway disease in infants and young children, but no safe and effective RSV vaccine is yet available.In the present study, we investigate suppressor of cytokine signaling (SOCS)-1 and SOCS3 expression associated with the type I IFN and IFN-stimulated gene (ISG)-15 response following infection of mouse lung epithelial (MLE-15) cells with RSV or RSV mutant viruses lacking the G gene, or NS1 and NS2 gene deletions.These results show an important role for SOCS1 regulation of the antiviral host response to RSV infection, and demonstrate a novel role for RSV G protein manipulation of SOCS3 and modulation of ISG15 and IFNbeta mRNA expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Infectious Diseases, Center for Disease Intervention, University of Georgia, Athens, GA 30602, USA. ecmoore@uga.edu

ABSTRACT
Respiratory syncytial virus (RSV) is a major cause of severe lower airway disease in infants and young children, but no safe and effective RSV vaccine is yet available. Factors attributing to this problem are associated with an incomplete understanding of the mechanisms by which RSV modulates the host cell response to infection. In the present study, we investigate suppressor of cytokine signaling (SOCS)-1 and SOCS3 expression associated with the type I IFN and IFN-stimulated gene (ISG)-15 response following infection of mouse lung epithelial (MLE-15) cells with RSV or RSV mutant viruses lacking the G gene, or NS1 and NS2 gene deletions. Studies in MLE-15 cells are important as this cell line represents the distal bronchiolar and alveolar epithelium of mice, the most common animal model used to evaluate the host cell response to RSV infection, and exhibit morphologic characteristics of alveolar type II cells, a primary cell type targeted during RSV infection. These results show an important role for SOCS1 regulation of the antiviral host response to RSV infection, and demonstrate a novel role for RSV G protein manipulation of SOCS3 and modulation of ISG15 and IFNbeta mRNA expression.

Show MeSH
Related in: MedlinePlus