scholarly journals Human coronaviruses 229E and OC43 replicate and induce distinct antiviral responses in differentiated primary human bronchial epithelial cells

2020 ◽  
Vol 319 (6) ◽  
pp. L926-L931
Author(s):  
Su-Ling Loo ◽  
Peter A. B. Wark ◽  
Camille Esneau ◽  
Kristy S. Nichol ◽  
Alan C-Y. Hsu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Time Course ◽  
Bronchial Epithelial Cells ◽  
Innate Immune ◽  
Infection Model ◽  
Type I ◽  
Human Bronchial Epithelial Cells ◽  
Cell Infection ◽  
Bronchial Epithelial ◽  
Human Coronaviruses

The recurrent emergence of novel, pathogenic coronaviruses (CoVs) severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1; 2002), Middle East respiratory syndrome (MERS)-CoV (2012), and most recently SARS-CoV-2 (2019) has highlighted the need for physiologically informative airway epithelial cell infection models for studying immunity to CoVs and development of antiviral therapies. To address this, we developed an in vitro infection model for two human coronaviruses; alphacoronavirus 229E-CoV (229E) and betacoronavirus OC43-CoV (OC43) in differentiated primary human bronchial epithelial cells (pBECs). Primary BECs from healthy subjects were grown at air-liquid interface (ALI) and infected with 229E or OC43, and replication kinetics and time-course expression of innate immune mediators were assessed. OC43 and 229E-CoVs replicated in differentiated pBECs but displayed distinct replication kinetics: 229E replicated rapidly with viral load peaking at 24 h postinfection, while OC43 replication was slower peaking at 96 h after infection. This was associated with diverse antiviral response profiles defined by increased expression of type I/III interferons and interferon-stimulated genes (ISGs) by 229E compared with no innate immune activation with OC43 infection. Understanding the host-virus interaction for previously established coronaviruses will give insight into pathogenic mechanisms underpinning SARS-CoV-2-induced respiratory disease and other future coronaviruses that may arise from zoonotic sources.

scholarly journals Asthmatic bronchial epithelial cells have a deficient innate immune response to infection with rhinovirus

2005 ◽  
Vol 201 (6) ◽  
pp. 937-947 ◽  
Author(s):  
Peter A.B. Wark ◽  
Sebastian L. Johnston ◽  
Fabio Bucchieri ◽  
Robert Powell ◽  
Sarah Puddicombe ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Virus Replication ◽  
Cellular Response ◽  
Bronchial Epithelial Cells ◽  
Innate Immune ◽  
Type I ◽  
Interferon Β ◽  
Bronchial Epithelial ◽  
Asthma Exacerbations ◽  
Viral Yield

Rhinoviruses are the major trigger of acute asthma exacerbations and asthmatic subjects are more susceptible to these infections. To investigate the underlying mechanisms of this increased susceptibility, we examined virus replication and innate responses to rhinovirus (RV)-16 infection of primary bronchial epithelial cells from asthmatic and healthy control subjects. Viral RNA expression and late virus release into supernatant was increased 50- and 7-fold, respectively in asthmatic cells compared with healthy controls. Virus infection induced late cell lysis in asthmatic cells but not in normal cells. Examination of the early cellular response to infection revealed impairment of virus induced caspase 3/7 activity and of apoptotic responses in the asthmatic cultures. Inhibition of apoptosis in normal cultures resulted in enhanced viral yield, comparable to that seen in infected asthmatic cultures. Examination of early innate immune responses revealed profound impairment of virus-induced interferon-β mRNA expression in asthmatic cultures and they produced >2.5 times less interferon-β protein. In infected asthmatic cells, exogenous interferon-β induced apoptosis and reduced virus replication, demonstrating a causal link between deficient interferon-β, impaired apoptosis and increased virus replication. These data suggest a novel use for type I interferons in the treatment or prevention of virus-induced asthma exacerbations.

scholarly journals Accelerated epithelial cell senescence in IPF and the inhibitory role of SIRT6 in TGF-β-induced senescence of human bronchial epithelial cells

2011 ◽  
Vol 300 (3) ◽  
pp. L391-L401 ◽  
Author(s):  
Shunsuke Minagawa ◽  
Jun Araya ◽  
Takanori Numata ◽  
Satoko Nojiri ◽  
Hiromichi Hara ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cellular Senescence ◽  
Type I Collagen ◽  
Transforming Growth Factor ◽  
Pathological Finding ◽  
Bronchial Epithelial Cells ◽  
Type I ◽  
Human Bronchial Epithelial Cells ◽  
Bronchial Epithelial ◽  
Expression Levels

Reepithelialization of remodeled air spaces with bronchial epithelial cells is a prominent pathological finding in idiopathic pulmonary fibrosis (IPF) and is implicated in IPF pathogenesis. Recent studies suggest that epithelial senescence is a risk factor for development of IPF, indicating such reepithelialization may be influenced by the acceleration of cellular senescence. Among the sirtuin (SIRT) family, SIRT6, a class III histone deacetylase, has been demonstrated to antagonize senescence. We evaluated the senescence of bronchiolization in association with SIRT6 expression in IPF lung. Senescence-associated β-galactosidase staining and immunohistochemical detection of p21 were performed to evaluate cellular senescence. As a model for transforming growth factor (TGF)-β-induced senescence of abnormal reepithelialization, we used primary human bronchial epithelial cells (HBEC). The changes of SIRT6, p21, and interleukin (IL)-1β expression levels in HBEC, as well as type I collagen expression levels in fibroblasts, were evaluated. In IPF lung samples, an increase in markers of senescence and SIRT6 expression was found in the bronchial epithelial cells lining cystically remodeled air spaces. We found that TGF-β induced senescence in primary HBEC by increasing p21 expression, and, whereas TGF-β also induced SIRT6, it was not sufficient to inhibit cellular senescence. However, overexpression of SIRT6 efficiently inhibited TGF-β-induced senescence via proteasomal degradation of p21. TGF-β-induced senescent HBEC secreted increased amounts of IL-1β, which was sufficient to induce myofibroblast differentiation in fibroblasts. These findings suggest that accelerated epithelial senescence plays a role in IPF pathogenesis through perpetuating abnormal epithelial-mesenchymal interactions, which can be antagonized by SIRT6.

Human bronchial epithelial cells can contract type I collagen gels

1998 ◽  
Vol 274 (1) ◽  
pp. L58-L65 ◽  
Author(s):  
Xiangde Liu ◽  
Takeshi Umino ◽  
Marty Cano ◽  
Ronald Ertl ◽  
Tom Veys ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Type I Collagen ◽  
Bronchial Epithelial Cells ◽  
Image Analyzer ◽  
Type I ◽  
Collagen Gels ◽  
Human Bronchial Epithelial Cells ◽  
Maximal Contraction ◽  
Bronchial Epithelial ◽  
Contract Type

Fibroblasts can contract collagen gels, a process thought to be related to tissue remodeling. Because epithelial cells are also involved in repair responses, we postulated that human bronchial epithelial cells (HBECs) could cause contraction of collagen gels. To evaluate this, HBECs were plated on the top of native type I collagen gels and were incubated for 48 h. After this, the gels were released and floated in LHC-9-RPMI 1640 for varying times, and gel size was measured with an image analyzer. HBECs caused a marked contraction of the gels within 24 h; the area was reduced by 88 ± 4% ( P < 0.01). The degree of gel contraction was dependent on cell density; 12,500 cells/cm2 resulted in maximal contraction, and half-maximal contraction occurred at 7,500 cells/cm2. Contraction varied inversely with the collagen concentration (91 ± 1% with 0.5 mg/ml collagen vs. 43 ± 5% with 1.5 mg/ml collagen). In contrast to fibroblasts that contract gels most efficiently when cast into the gel, HBEC-mediated contraction was significantly ( P < 0.01) more efficient when cells were on top of the gels rather than when cast into the gels. Anti-β1-integrin antibody blocked HBEC-mediated contraction by >50%, whereas anti-α2-, anti-α3-, anti-αv-, anti-αvβ5-, anti-β2-, or anti-β4-integrin antibody was without effect. The combination of anti-β1-integrin antibody and an anti-α-subfamily antibody completely blocked gel contraction induced by HBECs. In contrast, anti-cellular fibronectin antibody did not block HBEC-induced gel contraction, whereas it did block fibroblast-mediated gel contraction. In summary, human airway epithelial cells can contract type I collagen gels, a process that appears to require integrins but may not require fibronectin. This process may contribute to airway remodeling.

scholarly journals Human bronchial epithelial cells modulate collagen gel contraction by fibroblasts

1998 ◽  
Vol 274 (1) ◽  
pp. L119-L126 ◽  
Author(s):  
Tadashi Mio ◽  
Xiang-Der Liu ◽  
Yuichi Adachi ◽  
Ilja Striz ◽  
C. Magnus Sköld ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Conditioned Medium ◽  
Collagen Gel ◽  
Bronchial Epithelial Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Human Bronchial Epithelial Cells ◽  
Bronchial Epithelial ◽  
Tissue Contraction ◽  
Collagen Gel Contraction

Connective tissue contraction is an important aspect of both normal wound healing and fibrosis. This process may contribute to small airway narrowing associated with certain airway diseases. Fibroblast-mediated contraction of a three-dimensional collagen gel has been considered a model of tissue contraction. In this study, the ability of primary cultured human bronchial epithelial cells (HBEC) obtained by bronchial brushings to modulate fibroblast gel contraction was evaluated. Human lung fibroblasts (HFL1) were cast into type I collagen gels. The gels were floated both in dishes containing a monolayer of HBEC or in dishes without HBEC. Contraction assessed by measuring the area of gels was increased at all time points from 24 h up to 96 h of coculture. At 48 h, coculture of HBEC with fibroblasts resulted in significantly more contraction than fibroblasts alone (36.6 ± 1.2 vs. 20.4 ± 1.7%, P < 0.05). Lipopolysaccharide (LPS, 10 μg/ml) stimulation of the HBEC augmented the contraction (44.9 ± 1.0%, P < 0.05 vs. HBEC). In the presence of indomethacin, the augmentation by LPS was increased further (52.2 ± 4.3%, P< 0.05 vs. HBEC with LPS), suggesting that prostaglandins (PGs) are present and may inhibit contraction. Consistent with this, PGE was present in HBEC-conditioned medium. Bronchial epithelial cell conditioned medium had an effect similar to coculturing. SG-150 column chromatography revealed augmentive activity between 20 and 30 kDa and inhibitory activity between 10 and 20 kDa. Measurement by enzyme-linked immunosorbent assay confirmed the presence of the active form of transforming growth factor (TGF)-β2. The stimulatory activity of conditioned medium was blocked by adding anti-TGF-β antibody. These data demonstrate that, through the release of factors including TGF-β2 which can augment and PGE which can inhibit, HBEC can modulate fibroblast-mediated collagen gel contraction. In this manner, HBEC may modulate fibroblast activities that determine the architecture of bronchial tissue.

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