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Coyle, Peter; Ong, Grace; O’Neill, Hugh J.; McCaughey, Conall; Ornellas, Dennis De; Mitchell, F. E.; Mitchell, S.J.; Feeney, Susan; Wyatt, D.E.; Forde, Marian; Stockton, Joanne

doi:10.1186/1471-2180-4-41

Cited by 26 publications

(12 citation statements)

References 28 publications

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“…Fourteen uninfected and otherwise healthy children (mean age, 1.7 years; range, 1 to 2.4 years) acted as controls with blind nonbronchoscopic bronchoalveolar lavage samples obtained (after the instillation of 10 ml of saline) at the time of intubation for an elective surgical procedure (29). All RSV-infected infants were confirmed as having mono-infections by using a multiplex virus reverse transcriptase-PCR (RT-PCR) strip for 12 respiratory viruses, as previously described (30).…”

Section: Methodsmentioning

confidence: 99%

Induction and Antagonism of Antiviral Responses in Respiratory Syncytial Virus-Infected Pediatric Airway Epithelium

Villenave

Broadbent

Douglas³

et al. 2015

J Virol

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Airway epithelium is the primary target of many respiratory viruses. However, virus induction and antagonism of host responses by human airway epithelium remains poorly understood. To address this, we developed a model of respiratory syncytial virus (RSV) infection based on well-differentiated pediatric primary bronchial epithelial cell cultures (WD-PBECs) that mimics hallmarks of RSV disease in infants. RSV is the most important respiratory viral pathogen in young infants worldwide. We found that RSV induces a potent antiviral state in WD-PBECs that was mediated in part by secreted factors, including interferon lambda 1 ( IMPORTANCEMost respiratory viruses target airway epithelium for infection and replication, which is central to causing disease. However, for most human viruses we have a poor understanding of their interactions with human airway epithelium. Respiratory syncytial virus (RSV) is the most important viral pathogen of young infants. To help understand RSV interactions with pediatric airway epithelium, we previously developed three-dimensional primary cell cultures from infant bronchial epithelium that reproduce several hallmarks of RSV infection in infants, indicating that they represent authentic surrogates of RSV infection in infants. We found that RSV induced a potent antiviral state in these cultures and that a type III interferon, interleukin IL-29 (IL-29), was involved. Indeed, our data suggest that IL-29 has potential to prevent RSV disease. However, we also demonstrated that RSV efficiently circumvents this antiviral immune response and identified mechanisms by which this may occur. Our study provides new insights into RSV interaction with pediatric airway epithelium. Airway epithelium is an extremely important barrier to respiratory pathogens. It is also the primary infection target for many respiratory viruses. Elucidating the interactions between respiratory viruses and airway epithelium is fundamental to understanding aspects of their pathogenesis. We recently developed and characterized models of respiratory syncytial virus (RSV) infection based on well-differentiated pediatric primary airway epithelial cells derived from pediatric bronchial (WD-PBECs) or nasal (WD-PNECs) brushings (1, 2). RSV is the primary viral cause of infant hospitalizations in the first year of life and is capable of repeated infections throughout life (3). Despite its original isolation in 1957 (4), no effective RSV therapies or vaccines are available. The mechanisms by which RSV causes disease and is capable of repeated infections in humans remain an enigma. Our models reproduce several hallmarks of RSV infection in vivo, suggesting that they provide authentic surrogates with which to study RSVinduced innate immune responses and interaction with human airway epithelium (1, 2).We previously reported secretion of high levels of CXCL10, an

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Section: Methodsmentioning

confidence: 99%

Induction and Antagonism of Antiviral Responses in Respiratory Syncytial Virus-Infected Pediatric Airway Epithelium

Villenave

Broadbent

Douglas³

et al. 2015

J Virol

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“…Asthmatic PBECs and asthmatic PNECs were cultured as previously described [5] . All brush washings were analysed for viruses using a multi-viral PCR analysis and only uncontaminated cultures were used [22] .…”

Section: Methodsmentioning

confidence: 99%

Nasal Epithelial Cells Can Act as a Physiological Surrogate for Paediatric Asthma Studies

et al. 2014

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IntroductionDifferentiated paediatric epithelial cells can be used to study the role of epithelial cells in asthma. Nasal epithelial cells are easier to obtain and may act as a surrogate for bronchial epithelium in asthma studies. We assessed the suitability of nasal epithelium from asthmatic children to be a surrogate for bronchial epithelium using air-liquid interface cultures.MethodsPaired nasal and bronchial epithelial cells from asthmatic children (n = 9) were differentiated for 28 days under unstimulated and IL-13-stimulated conditions. Morphological and physiological markers were analysed using immunocytochemistry, transepithelial-electrical-resistance, Quantitative Real-time-PCR, ELISA and multiplex cytokine/chemokine analysis.ResultsPhysiologically, nasal epithelial cells from asthmatic children exhibit similar cytokine responses to stimulation with IL-13 compared with paired bronchial epithelial cells. Morphologically however, nasal epithelial cells differed significantly from bronchial epithelial cells from asthmatic patients under unstimulated and IL-13-stimulated conditions. Nasal epithelial cells exhibited lower proliferation/differentiation rates and lower percentages of goblet and ciliated cells when unstimulated, while exhibiting a diminished and varied response to IL-13.ConclusionsWe conclude that morphologically, nasal epithelial cells would not be a suitable surrogate due to a significantly lower rate of proliferation and differentiation of goblet and ciliated cells. Physiologically, nasal epithelial cells respond similarly to exogenous stimulation with IL-13 in cytokine production and could be used as a physiological surrogate in the event that bronchial epithelial cells are not available.

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“…This represents a culture success rate of 70% of PBEC samples received. All brush washings were analysed for viruses using a multi-viral PCR analysis and only uncontaminated cultures were used [26].…”

Section: Methodsmentioning

confidence: 99%

Chronic IL9 and IL-13 Exposure Leads to an Altered Differentiation of Ciliated Cells in a Well-Differentiated Paediatric Bronchial Epithelial Cell Model

et al. 2013

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Asthma is a chronic inflammatory disease characterised by airways remodelling. In mouse models IL-9 and IL-13 have been implicated in airways remodelling including mucus hypersecretion and goblet cell hyperplasia. Their role, especially that of IL-9, has been much less studied in authentic human ex vivo models of the bronchial epithelium from normal and asthmatic children. We assessed the effects of IL-9, IL-13 and an IL-9/IL-13 combination, during differentiation of bronchial epithelial cells from normal (n = 6) and asthmatic (n = 8) children. Cultures were analysed for morphological markers and factors associated with altered differentiation (MUC5AC, SPDEF and MMP-7). IL-9, IL-9/IL-13 combination and IL-13 stimulated bronchial epithelial cells from normal children had fewer ciliated cells [14.8% (SD 8.9), p = 0.048, 12.4 (SD 6.1), p = 0.016 and 7.3% (SD 6.6), p = 0.031] respectively compared with unstimulated [(21.4% (SD 9.6)]. IL-9 stimulation had no effect on goblet cell number in either group whereas IL-9/IL-13 combination and IL-13 significantly increased goblet cell number [24.8% (SD 8.8), p = 0.02), 32.9% (SD 8.6), p = 0.007] compared with unstimulated normal bronchial cells [(18.6% (SD 6.2)]. All stimulations increased MUC5AC mRNA in bronchial epithelial cells from normal children and increased MUC5AC mucin secretion. MMP-7 localisation was dysregulated in normal bronchial epithelium stimulated with Th2 cytokines which resembled the unstimulated bronchial epithelium of asthmatic children. All stimulations resulted in a significant reduction in transepithelial electrical resistance values over time suggesting a role in altered tight junction formation. We conclude that IL-9 does not increase goblet cell numbers in bronchial epithelial cell cultures from normal or asthmatic children. IL-9 and IL-13 alone and in combination, reduce ciliated cell numbers and transepithelial electrical resistance during differentiation of normal epithelium, which clinically could inhibit mucociliary clearance and drive an altered repair mechanism. This suggests an alternative role for IL-9 in airways remodelling and reaffirms IL-9 as a potential therapeutic target.

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Cited by 26 publications

References 28 publications

Induction and Antagonism of Antiviral Responses in Respiratory Syncytial Virus-Infected Pediatric Airway Epithelium

Induction and Antagonism of Antiviral Responses in Respiratory Syncytial Virus-Infected Pediatric Airway Epithelium

Nasal Epithelial Cells Can Act as a Physiological Surrogate for Paediatric Asthma Studies

Chronic IL9 and IL-13 Exposure Leads to an Altered Differentiation of Ciliated Cells in a Well-Differentiated Paediatric Bronchial Epithelial Cell Model

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