scholarly journals Susceptibility of primary human airway epithelial cells to Bordetella pertussis adenylate cyclase toxin in two- and three-dimensional culture conditions

2020 ◽  
Vol 27 (1) ◽  
pp. 89-98
Author(s):  
Maria Bianchi ◽  
Rinu Sivarajan ◽  
Thorsten Walles ◽  
Stephan Hackenberg ◽  
Maria Steinke
Keyword(s):  
Epithelial Cells ◽  
Adenylate Cyclase ◽  
Cell Lines ◽  
Bordetella Pertussis ◽  
Three Dimensional ◽  
Airway Epithelial Cells ◽  
Culture Conditions ◽  
Airway Epithelial ◽  
Human Airway ◽  
The Impact

The human pathogen Bordetella pertussis targets the respiratory epithelium and causes whooping cough. Its virulence factor adenylate cyclase toxin (CyaA) plays an important role in the course of infection. Previous studies on the impact of CyaA on human epithelial cells have been carried out using cell lines derived from the airways or the intestinal tract. Here, we investigated the interaction of CyaA and its enzymatically inactive but fully pore-forming toxoid CyaA-AC– with primary human airway epithelial cells (hAEC) derived from different anatomical sites (nose and tracheo-bronchial region) in two-dimensional culture conditions. To assess possible differences between the response of primary hAEC and respiratory cell lines directly, we included HBEC3-KT in our studies. In comparative analyses, we studied the impact of both the toxin and the toxoid on cell viability, intracellular cAMP concentration and IL-6 secretion. We found that the selected hAEC, which lack CD11b, were differentially susceptible to both CyaA and CyaA-AC–. HBEC3-KT appeared not to be suitable for subsequent analyses. Since the nasal epithelium first gets in contact with airborne pathogens, we further studied the effect of CyaA and its toxoid on the innate immunity of three-dimensional tissue models of the human nasal mucosa. The present study reveals first insights in toxin–cell interaction using primary hAEC.

scholarly journals Choice of Differentiation Media Significantly Impacts Cell Lineage and Response to CFTR Modulators in Fully Differentiated Primary Cultures of Cystic Fibrosis Human Airway Epithelial Cells

Cells ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 2137
Author(s):  
Vinciane Saint-Criq ◽  
Livia Delpiano ◽  
John Casement ◽  
Jennifer C. Onuora ◽  
JinHeng Lin ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Epithelial Cells ◽  
Cell Lineage ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Functional Responses ◽  
Human Airway ◽  
The Impact ◽  
Cftr Modulators ◽  
Human Airway Epithelial Cells

In vitro cultures of primary human airway epithelial cells (hAECs) grown at air–liquid interface have become a valuable tool to study airway biology under normal and pathologic conditions, and for drug discovery in lung diseases such as cystic fibrosis (CF). An increasing number of different differentiation media, are now available, making comparison of data between studies difficult. Here, we investigated the impact of two common differentiation media on phenotypic, transcriptomic, and physiological features of CF and non-CF epithelia. Cellular architecture and density were strongly impacted by the choice of medium. RNA-sequencing revealed a shift in airway cell lineage; one medium promoting differentiation into club and goblet cells whilst the other enriched the growth of ionocytes and multiciliated cells. Pathway analysis identified differential expression of genes involved in ion and fluid transport. Physiological assays (intracellular/extracellular pH, Ussing chamber) specifically showed that ATP12A and CFTR function were altered, impacting pH and transepithelial ion transport in CF hAECs. Importantly, the two media differentially affected functional responses to CFTR modulators. We argue that the effect of growth conditions should be appropriately determined depending on the scientific question and that our study can act as a guide for choosing the optimal growth medium for specific applications.

scholarly journals The impact of oil spill to lung health—Insights from an RNA-seq study of human airway epithelial cells

Gene ◽  
2016 ◽  
Vol 578 (1) ◽  
pp. 38-51 ◽  
Author(s):  
Yao-Zhong Liu ◽  
Astrid M. Roy-Engel ◽  
Melody C. Baddoo ◽  
Erik K. Flemington ◽  
Guangdi Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Oil Spill ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Rna Seq ◽  
Human Airway ◽  
Lung Health ◽  
The Impact ◽  
Human Airway Epithelial Cells

scholarly journals Assessment of the infectious threshold of SARS-CoV-2 in primary airway epithelial cells

2021 ◽  
Author(s):  
Manel ESSAIDI-LAZIOSI ◽  
Francisco Javier Perez Rodriguez ◽  
Pascale Sattonnet Roche ◽  
Nicolas Hulo ◽  
Frederique Jacquerioz ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Lines ◽  
Virus Isolation ◽  
Airway Epithelial Cells ◽  
Clinical Samples ◽  
Airway Epithelial ◽  
Airway Epithelia ◽  
Viral Loads ◽  
Human Airway ◽  
Human Airway Epithelia

Comparison of virus isolation success from clinical samples across a range of viral loads inoculated in parallel on Vero E6 and human airway epithelia (HAE) showed lower success of virus isolation in HAE, suggesting an overestimation of actual infectiousness in humans using Vero E6 cell lines, commonly considered as reference.

A Toolbox for Studying Respiratory Viral Infections Using Air-Liquid Interface Cultures of Human Airway Epithelial Cells

2021 ◽  
Author(s):  
Aubrey Nicole Michi ◽  
David Proud
Keyword(s):  
Epithelial Cells ◽  
Cell Lines ◽  
Viral Infections ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Human Airway ◽  
Respiratory Viral Infections ◽  
Air Liquid Interface ◽  
Human Airway Epithelial Cells

Submerged cultures of primary human airway epithelial cells, or human airway epithelial cell lines have been a mainstay of airway epithelial biology research for decades due to their robust in vitro proliferative capacity, relatively low maintenance culture conditions, and clinically translatable results to nasal or bronchial brushings. With the development and improvement of air-liquid interface (ALI) cultures of human airway epithelial cells, such cultures have been considered superior to immortalized cell lines and primary cell monolayers as such cultures effectively recapitulate in vivo epithelial architecture and cell types. Although ALI culture growth protocols are well-established and widely available, many researchers have avoided their use, as ALI cultures not only take longer to grow but also present technical challenges and limitations that make in vitro intracellular and structural assays taxing. Challenges arise relating to their complex structure, requirements for air exposure, the constraints of transwell growth apparatus, and interference in assays caused by mucus secretion. Although few publications briefly describe technical adaptations for some assays, there is still considerable trial and error required for researchers to establish consistent and reliable assay adaptations, often becoming a deterrent for pursuing mechanistic investigation. We have created a user-friendly toolbox detailing comprehensive protocols for numerous techniques and assay adaptations, particularly focusing on respiratory virus infections. By expanding the repertoire of ALI culture-adapted in vitro assays, we hope to facilitate the widespread adoption of this valuable culture system for mechanistic investigations of respiratory viral infections or other epithelial-pathogen models.

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