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Trehalose-mediated autophagy impairs the anti-viral function of human primary airway epithelial cells.

Wu Q, Jiang D, Huang C, van Dyk LF, Li L, Chu HW - PLoS ONE (2015)

Bottom Line: We found that trehalose-induced autophagy significantly impaired IFN-λ1 expression and increased HRV-16 load.Inhibition of autophagy via knockdown of autophagy-related gene 5 (ATG5) effectively rescued the impaired IFN-λ1 expression by trehalose and subsequently reduced HRV-16 load.Intervention of excessive autophagy in chronic lung diseases may provide a novel approach to attenuate viral infections and associated disease exacerbations.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, National Jewish Health, Denver, Colorado, United States of America.

ABSTRACT
Human rhinovirus (HRV) is the most common cause of acute exacerbations of chronic lung diseases including asthma. Impaired anti-viral IFN-λ1 production and increased HRV replication in human asthmatic airway epithelial cells may be one of the underlying mechanisms leading to asthma exacerbations. Increased autophagy has been shown in asthmatic airway epithelium, but the role of autophagy in anti-HRV response remains uncertain. Trehalose, a natural glucose disaccharide, has been recognized as an effective autophagy inducer in mammalian cells. In the current study, we used trehalose to induce autophagy in normal human primary airway epithelial cells in order to determine if autophagy directly regulates the anti-viral response against HRV. We found that trehalose-induced autophagy significantly impaired IFN-λ1 expression and increased HRV-16 load. Inhibition of autophagy via knockdown of autophagy-related gene 5 (ATG5) effectively rescued the impaired IFN-λ1 expression by trehalose and subsequently reduced HRV-16 load. Mechanistically, ATG5 protein interacted with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1), two critical molecules involved in the expression of anti-viral interferons. Our results suggest that induction of autophagy in human primary airway epithelial cells inhibits the anti-viral IFN-λ1 expression and facilitates HRV infection. Intervention of excessive autophagy in chronic lung diseases may provide a novel approach to attenuate viral infections and associated disease exacerbations.

No MeSH data available.


Related in: MedlinePlus

Autophagy-related gene 5 (ATG5) protein interacts with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1) in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were treated with medium or trehalose (TRE, 100 mM) for 48 h and then infected with HRV-16 (104 TCID50/well) for 2 h. After removing the free viruses, cells were incubated with medium or trehalose for additional 6 h. Pre-cleared cell lysates were incubated with a mouse anti-human ATG5 antibody, and immunoprecipitated proteins were separated on 10% SDS—PAGE for immunoblotting of RIG-I, IPS-1 and ATG5. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells.
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pone.0124524.g006: Autophagy-related gene 5 (ATG5) protein interacts with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1) in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were treated with medium or trehalose (TRE, 100 mM) for 48 h and then infected with HRV-16 (104 TCID50/well) for 2 h. After removing the free viruses, cells were incubated with medium or trehalose for additional 6 h. Pre-cleared cell lysates were incubated with a mouse anti-human ATG5 antibody, and immunoprecipitated proteins were separated on 10% SDS—PAGE for immunoblotting of RIG-I, IPS-1 and ATG5. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells.

Mentions: To uncover the potential molecular mechanisms involved in autophagy-mediated suppression of IFN-λ1, the interaction of ATG5 protein with RIG-I and IPS-1 was examined by ATG5 pull-down, followed by immunoblotting of RIG-I and IPS-1 in NHTE cells after treatment with or without trehalose and HRV-16 for 6 h. ATG5 protein was constitutively expressed at a low level in cultured NHTE cells. RIG-I and IPS-1 were co-immunoprecipitated with ATG5 protein (Fig 6). A stronger interaction of RIG-I and IPS-1 with ATG5 protein was observed in trehalose-treated cells, especially with HRV-16 infection. These observations indicate that ATG5 may interact with RIG-I and IPS-1 upon induction of autophagy in normal human primary airway epithelial cells.


Trehalose-mediated autophagy impairs the anti-viral function of human primary airway epithelial cells.

Wu Q, Jiang D, Huang C, van Dyk LF, Li L, Chu HW - PLoS ONE (2015)

Autophagy-related gene 5 (ATG5) protein interacts with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1) in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were treated with medium or trehalose (TRE, 100 mM) for 48 h and then infected with HRV-16 (104 TCID50/well) for 2 h. After removing the free viruses, cells were incubated with medium or trehalose for additional 6 h. Pre-cleared cell lysates were incubated with a mouse anti-human ATG5 antibody, and immunoprecipitated proteins were separated on 10% SDS—PAGE for immunoblotting of RIG-I, IPS-1 and ATG5. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4400043&req=5

pone.0124524.g006: Autophagy-related gene 5 (ATG5) protein interacts with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1) in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were treated with medium or trehalose (TRE, 100 mM) for 48 h and then infected with HRV-16 (104 TCID50/well) for 2 h. After removing the free viruses, cells were incubated with medium or trehalose for additional 6 h. Pre-cleared cell lysates were incubated with a mouse anti-human ATG5 antibody, and immunoprecipitated proteins were separated on 10% SDS—PAGE for immunoblotting of RIG-I, IPS-1 and ATG5. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells.
Mentions: To uncover the potential molecular mechanisms involved in autophagy-mediated suppression of IFN-λ1, the interaction of ATG5 protein with RIG-I and IPS-1 was examined by ATG5 pull-down, followed by immunoblotting of RIG-I and IPS-1 in NHTE cells after treatment with or without trehalose and HRV-16 for 6 h. ATG5 protein was constitutively expressed at a low level in cultured NHTE cells. RIG-I and IPS-1 were co-immunoprecipitated with ATG5 protein (Fig 6). A stronger interaction of RIG-I and IPS-1 with ATG5 protein was observed in trehalose-treated cells, especially with HRV-16 infection. These observations indicate that ATG5 may interact with RIG-I and IPS-1 upon induction of autophagy in normal human primary airway epithelial cells.

Bottom Line: We found that trehalose-induced autophagy significantly impaired IFN-λ1 expression and increased HRV-16 load.Inhibition of autophagy via knockdown of autophagy-related gene 5 (ATG5) effectively rescued the impaired IFN-λ1 expression by trehalose and subsequently reduced HRV-16 load.Intervention of excessive autophagy in chronic lung diseases may provide a novel approach to attenuate viral infections and associated disease exacerbations.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, National Jewish Health, Denver, Colorado, United States of America.

ABSTRACT
Human rhinovirus (HRV) is the most common cause of acute exacerbations of chronic lung diseases including asthma. Impaired anti-viral IFN-λ1 production and increased HRV replication in human asthmatic airway epithelial cells may be one of the underlying mechanisms leading to asthma exacerbations. Increased autophagy has been shown in asthmatic airway epithelium, but the role of autophagy in anti-HRV response remains uncertain. Trehalose, a natural glucose disaccharide, has been recognized as an effective autophagy inducer in mammalian cells. In the current study, we used trehalose to induce autophagy in normal human primary airway epithelial cells in order to determine if autophagy directly regulates the anti-viral response against HRV. We found that trehalose-induced autophagy significantly impaired IFN-λ1 expression and increased HRV-16 load. Inhibition of autophagy via knockdown of autophagy-related gene 5 (ATG5) effectively rescued the impaired IFN-λ1 expression by trehalose and subsequently reduced HRV-16 load. Mechanistically, ATG5 protein interacted with retinoic acid-inducible gene I (RIG-I) and IFN-β promoter stimulator 1 (IPS-1), two critical molecules involved in the expression of anti-viral interferons. Our results suggest that induction of autophagy in human primary airway epithelial cells inhibits the anti-viral IFN-λ1 expression and facilitates HRV infection. Intervention of excessive autophagy in chronic lung diseases may provide a novel approach to attenuate viral infections and associated disease exacerbations.

No MeSH data available.


Related in: MedlinePlus