scholarly journals Rhinovirus C replication is associated with the endoplasmic reticulum and triggers cytopathic effects in an in vitro model of human airway epithelium

2022 ◽  
Vol 18 (1) ◽  
pp. e1010159
Author(s):  
Talita B. Gagliardi ◽  
Monty E. Goldstein ◽  
Daniel Song ◽  
Kelsey M. Gray ◽  
Jae W. Jung ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Airway Epithelium ◽  
High Ratio ◽  
Cytopathic Effects ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Rhinovirus C ◽  
The Impact

The clinical impact of rhinovirus C (RV-C) is well-documented; yet, the viral life cycle remains poorly defined. Thus, we characterized RV-C15 replication at the single-cell level and its impact on the human airway epithelium (HAE) using a physiologically-relevant in vitro model. RV-C15 replication was restricted to ciliated cells where viral RNA levels peaked at 12 hours post-infection (hpi), correlating with elevated titers in the apical compartment at 24hpi. Notably, infection was associated with a loss of polarized expression of the RV-C receptor, cadherin-related family member 3. Visualization of double-stranded RNA (dsRNA) during RV-C15 replication revealed two distinct replication complex arrangements within the cell, likely corresponding to different time points in infection. To further define RV-C15 replication sites, we analyzed the expression and colocalization of giantin, phosphatidylinositol-4-phosphate, and calnexin with dsRNA. Despite observing Golgi fragmentation by immunofluorescence during RV-C15 infection as previously reported for other RVs, a high ratio of calnexin-dsRNA colocalization implicated the endoplasmic reticulum as the primary site for RV-C15 replication in HAE. RV-C15 infection was also associated with elevated stimulator of interferon genes (STING) expression and the induction of incomplete autophagy, a mechanism used by other RVs to facilitate non-lytic release of progeny virions. Notably, genetic depletion of STING in HAE attenuated RV-C15 and -A16 (but not -B14) replication, corroborating a previously proposed proviral role for STING in some RV infections. Finally, RV-C15 infection resulted in a temporary loss in epithelial barrier integrity and the translocation of tight junction proteins while a reduction in mucociliary clearance indicated cytopathic effects on epithelial function. Together, our findings identify both shared and unique features of RV-C replication compared to related rhinoviruses and define the impact of RV-C on both epithelial cell organization and tissue functionality–aspects of infection that may contribute to pathogenesis in vivo.

scholarly journals Rhinovirus C replication is associated with the endoplasmic reticulum and triggers cytopathic effects in an in vitro model of human airway epithelium

2021 ◽  
Author(s):  
Talita B Gagliardi ◽  
Monty E Goldstein ◽  
Daniel Song ◽  
Kelsey M Gray ◽  
Jae W Jung ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Airway Epithelium ◽  
In Vitro Model ◽  
Cytopathic Effects ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Vitro Model ◽  
Rhinovirus C ◽  
The Impact

The clinical impact of rhinovirus C (RV-C) is well-documented; yet the viral life cycle remains poorly defined. Thus, we characterized RV-C15 replication at the single-cell level and its impact on the human airway epithelium (HAE) using a physiologically-relevant in vitro model. RV-C15 replication was restricted to ciliated cells where viral RNA levels peaked at 12 hours post-infection (hpi), correlating with elevated titers in the apical compartment at 24 hpi. Notably, infection was associated with a loss of polarized expression of the RV-C receptor, cadherin-related family member 3. Visualization of double-stranded RNA (dsRNA) during RV-C15 replication revealed two distinct replication complex arrangements within the cell, likely corresponding to different time points in infection and correlating with the formation of large intracellular vesicles. To further define RV-C15 replication sites, we analyzed the expression of giantin, phosphatidylinositol-4-phosphate, and calnexin, as well as the colocalization of these markers with dsRNA. Fluorescence levels of all three cellular markers were elevated during infection and altered giantin distribution further indicated Golgi fragmentation. However, unlike previously characterized RVs, the high ratio of calnexin-dsRNA colocalization implicated the endoplasmic reticulum as the primary site for RV-C15 replication in HAE. RV-C15 infection was also associated with elevated stimulator of interferon genes (STING) expression, facilitating replication, and the induction of incomplete autophagy, a mechanism used by other RVs to promote non-lytic release of progeny virions. Finally, RV-C15 infection resulted in a temporary loss in epithelial barrier integrity and the translocation of tight junction proteins while a reduction in mucociliary clearance indicated cytopathic effects on epithelial function. Together, our findings identify both shared and unique features of RV-C replication compared to related rhinoviruses and define the impact of RV-C on both epithelial cell organization and tissue functionality - aspects of infection that may contribute to pathogenesis in vivo.

scholarly journals An approach to testing undiluted e-cigarette aerosol in vitro using 3D reconstituted human airway epithelium

2019 ◽  
Vol 54 ◽  
pp. 391-401 ◽  
Author(s):  
E. Bishop ◽  
L. Haswell ◽  
J. Adamson ◽  
S. Costigan ◽  
D. Thorne ◽  
...  
Keyword(s):  
Airway Epithelium ◽  
Human Airway ◽  
Human Airway Epithelium

An in vitro model of human airway epithelium (EpiAirway) for in vitro metabolism and toxicity screening

2007 ◽  
Vol 172 ◽  
pp. S79 ◽  
Author(s):  
Patrick Hayden ◽  
Joseph Kubilus ◽  
Helena Kandárová ◽  
Mitchell Klausner ◽  
George Jackson ◽  
...  
Keyword(s):  
Airway Epithelium ◽  
In Vitro Model ◽  
In Vitro Metabolism ◽  
Toxicity Screening ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Vitro Model

scholarly journals DAS181, a sialidase fusion protein, protects human airway epithelium against influenza virus infection: an in vitro pharmacodynamic analysis

2009 ◽  
Vol 65 (2) ◽  
pp. 275-284 ◽  
Author(s):  
Gallen B. Triana-Baltzer ◽  
Maria Babizki ◽  
Michael C. W. Chan ◽  
Adam C. N. Wong ◽  
Laura M. Aschenbrenner ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Virus Infection ◽  
Fusion Protein ◽  
Airway Epithelium ◽  
Influenza Virus Infection ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Pharmacodynamic Analysis

In vivo antioxidant gene expression in human airway epithelium of normal individuals exposed to 100% O2

1993 ◽  
Vol 75 (3) ◽  
pp. 1256-1262 ◽  
Author(s):  
S. C. Erzurum ◽  
C. Danel ◽  
A. Gillissen ◽  
C. S. Chu ◽  
B. C. Trapnell ◽  
...  
Keyword(s):  
Gene Expression ◽  
Airway Epithelium ◽  
Oxidant Stress ◽  
Mrna Level ◽  
Bronchial Epithelium ◽  
Antioxidant Gene ◽  
Human Airway ◽  
Human Airway Epithelium ◽  
Antioxidant Gene Expression

Human bronchial epithelium is exquisitely sensitive to high O2 levels, with tracheobronchitis usually developing after 12 h of exposure to 100% O2. To evaluate whether this vulnerability results from inability of the bronchial epithelium to provide adequate antioxidant protection, we quantified antioxidant gene expression in bronchial epithelium of normal volunteers at baseline and after exposure to 100% O2 in vivo. After 14.8 +/- 0.2 h of 100% O2, 24 of 33 individuals had evidence of tracheobronchitis. Baseline gene expression of CuZn superoxide dismutase (SOD), MnSOD, and catalase in bronchial epithelium was very low (CuZnSOD 4.1 +/- 0.8 transcripts/cell, MnSOD 5.1 +/- 0.9, catalase 1.3 +/- 0.2), with control gamma-actin expression relatively abundant (50 +/- 6 transcripts/cell). Importantly, despite 100% O2 exposure sufficient to cause tracheobronchitis in most individuals, antioxidant mRNA transcripts/cell in bronchial epithelium did not increase (P > 0.5). Catalase activity in bronchial epithelium did not change after exposure to hyperoxia (P > 0.05). Total SOD activity increased mildly (P < 0.01) but not sufficiently to protect the epithelium. Together, the very low levels of expression of intracellular antioxidant enzymes and the inability to upregulate expression at the mRNA level with oxidant stress likely have a role in human airway epithelium susceptibility to hyperoxia.

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