A differentiated porcine bronchial epithelial cell culture model for studying human adenovirus tropism and virulence

Lam, Elena; Ramke, Mirja; Groos, Stephanie; Warnecke, G.; Heim, Albert

doi:10.1016/j.jviromet.2011.08.025

Cited by 26 publications

(28 citation statements)

References 20 publications

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“…The growth conditions of swALI cultures were different from those described for human ALI cultures. The difficulty in establishing this system may explain why so far only few reports about differentiated respiratory epithelial cells of swine are available161718. The novel feature in our study is that we analysed events occurring late after infection not addressed in previous analyses with swine or human ALI cultures.…”

Section: Discussionmentioning

confidence: 99%

“…PTEC and PBEC were obtained from swine trachea and bronchi, respectively. Primary cells were harvested as previously described18 and were seeded on type I collagen (Sigma)-coated flasks with bronchial epithelial cell serum-free growth medium (BEGM). The BEGM was modified from previous studies2829 and contained the BEBM basal medium (Lonza) supplemented with the required additives.…”

Section: Methodsmentioning

confidence: 99%

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The differentiated airway epithelium infected by influenza viruses maintains the barrier function despite a dramatic loss of ciliated cells

Yang

Beineke³

et al. 2016

Sci Rep

108

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Virus-host interactions in the respiratory epithelium during long term influenza virus infection are not well characterized. Therefore, we developed an air-liquid interface culture system for differentiated porcine respiratory epithelial cells to study the effect of virus-induced cellular damage. In our well-differentiated cells, α2,6-linked sialic acid is predominantly expressed on the apical surface and the basal cells mainly express α2,3-linked sialic acid. During the whole infection period, release of infectious virus was maintained at a high titre for more than seven days. The infected epithelial cells were subject to apoptosis resulting in the loss of ciliated cells together with a thinner thickness. Nevertheless, the airway epithelium maintained trans-epithelial electrical resistance and retained its barrier function. The loss of ciliated cells was compensated by the cells which contained the KRT5 basal cell marker but were not yet differentiated into ciliated cells. These specialized cells showed an increase of α2,3-linked sialic acid on the apical surface. In sum, our results help to explain the localized infection of the airway epithelium by influenza viruses. The impairment of mucociliary clearance in the epithelial cells provides an explanation why prior viral infection renders the host more susceptible to secondary co-infection by another pathogen.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The differentiated airway epithelium infected by influenza viruses maintains the barrier function despite a dramatic loss of ciliated cells

Yang

Beineke³

et al. 2016

Sci Rep

108

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“…ALI cultures from porcine airway cells have only recently been described for infection studies with viruses such as influenza virus and adenovirus2338 but not yet for bacterial infections. S. suis had been analyzed using a variety of immortalized respiratory epithelial cells1231323942, as well as polar porcine choroid plexus epithelial cells and human choroid plexus papilloma cells4546.…”

Section: Discussionmentioning

confidence: 99%

Efficient suilysin-mediated invasion and apoptosis in porcine respiratory epithelial cells after streptococcal infection under air-liquid interface conditions

Meng

Seitz

et al. 2016

Sci Rep

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Streptococci may colonize the epithelium in the airways and other entry sites. While local infection often remains asymptomatic, severe or even fatal diseases occur when streptococci become invasive and spread to different sites in the infected host. We have established porcine respiratory air-liquid interface cultures (ALI) from the porcine lung to analyze the interaction of streptococci with their primary target cells. As representative of the streptococcal family we chose Streptococcus suis (S. suis) that is not only a major swine respiratory pathogen but can also infect humans. Suilysin, a cholesterol-dependent cytolysin (CDC), is an important virulence factor. By comparing a S. suis wt strain with a suilysin-deficient mutant, we demonstrate that suilysin contributes to (i) adherence to airway cells (ii) loss of ciliated cells (iii) apoptosis, and (iv) invasion. Furthermore, we show that cytolytic activity of suilysin is crucial for these effects. A striking result of our analysis was the high efficiency of S. suis-induced apoptosis and invasion upon infection under ALI conditions. These properties have been reported to be less efficient when analyzed with immortalized cells. We hypothesize that soluble effectors such as suilysin are present at higher concentrations in cells kept at ALI conditions and thus more effective. These results should be relevant also for infection of the respiratory tract by other respiratory pathogens.

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“…Most primary cells senesce within a few passages; additionally, these cells can undergo spontaneous transdifferentiation into other epithelial cell types 17 or undergo epithelial-mesenchymal transition owing to the autocrine production of transforming growth factor-β (TGF-β) in culture [18][19][20] . To circumvent this, many studies have used immortalized primary epithelial cells 21,22 , established cell lines derived from tumors 23 or cells isolated from large animals 24 to study disease mechanisms or regeneration. With the development of iPSCs, tissue-specific cells generated from iPSCs hold great promise for patient-specific disease modeling, drug discovery and personalized medicine.…”

Section: Introductionmentioning

confidence: 99%

Efficient generation of functional CFTR-expressing airway epithelial cells from human pluripotent stem cells

et al. 2015

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Airway epithelial cells are of great interest for research on lung development, regeneration and disease modeling. This protocol describes how to generate cystic fibrosis (CF) transmembrane conductance regulator protein (CFTR)-expressing airway epithelial cells from human pluripotent stem cells (PSCs). The stepwise approach from PSC culture to differentiation into progenitors and then mature epithelia with apical CFTR activity is outlined. Human PSCs that were inefficient at endoderm differentiation using our previous lung differentiation protocol were able to generate substantial lung progenitor cell populations. Augmented CFTR activity can be observed in all cultures as early as at 35 d of differentiation, and full maturation of the cells in air-liquid interface cultures occurs in <5 weeks. This protocol can be used for drug discovery, tissue regeneration or disease modeling.

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A differentiated porcine bronchial epithelial cell culture model for studying human adenovirus tropism and virulence

Cited by 26 publications

References 20 publications

The differentiated airway epithelium infected by influenza viruses maintains the barrier function despite a dramatic loss of ciliated cells

The differentiated airway epithelium infected by influenza viruses maintains the barrier function despite a dramatic loss of ciliated cells

Efficient suilysin-mediated invasion and apoptosis in porcine respiratory epithelial cells after streptococcal infection under air-liquid interface conditions

Efficient generation of functional CFTR-expressing airway epithelial cells from human pluripotent stem cells

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