Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cozens, Daniel; Sutherland, Erin; Marchesi, Francesco; Taylor, Geraldine; Berry, Catherine C.; Davies, Robert L.

doi:10.1038/s41598-018-33180-w

Cited by 24 publications

(33 citation statements)

References 92 publications

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“…Another study also showed that the TEER of human nasal epithelial cells cultured in a LLI system dropped to approximately 500 Ω.cm 2 after reaching the maximum TEER value (3,133 ± 665 Ω.cm 2)45 . The drop in TEER values before stabilization could be related to the beginning of cell senescence 46 or differentiation state of the cells 43 .…”

Section: Discussionmentioning

confidence: 99%

Mannheimia haemolytica and lipopolysaccharide induce airway epithelial inflammatory responses in an extensively developed ex vivo calf model

Cai

Varasteh

Putten

et al. 2020

Sci Rep

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Pulmonary infection is associated with inflammation and damage to the bronchial epithelium characterized by an increase in the release of inflammatory factors and a decrease in airway barrier function. Our objective is to optimize a method for the isolation and culture of primary bronchial epithelial cells (PBECs) and to provide an ex vivo model to study mechanisms of epithelial airway inflammation. PBECs were isolated and cultured from the airways of calves in a submerged cell culture and liquid-liquid interface system. A higher yield and cell viability were obtained after stripping the epithelium from the bronchial section compared to cutting the bronchial section in smaller pieces prior to digestion. Mannheimia haemolytica and lipopolysaccharide (LPS) as stimulants increased inflammatory responses (IL-8, IL-6 and TNF-α release), possibly, by the activation of "TLR-mediated MAPKs and NF-κB" signaling. Furthermore, M. haemolytica and LpS disrupted the bronchial epithelial layer as observed by a decreased transepithelial electrical resistance and zonula occludens-1 and E-cadherin expression. An optimized isolation and culture method for calf PBECs was developed, which cooperated with animal use Replacement, Reduction and Refinement (3R's) principle, and can also contribute to the increased knowledge and development of effective therapies for other animal and humans (childhood) respiratory diseases. Respiratory diseases, such as pneumonia and bronchiolitis, are complex, multifactorial disorders caused by viral and/or microbial pathogens, an impaired immune system, as well as environmental and genetic factors. It is generally accepted that inflammation and damage of bronchial epithelium contribute to the development of respiratory diseases in animals and also humans 1-6. The airway epithelium plays a central role in the maintenance of airway integrity and acts as a physical barrier to protect the lungs against inhaled (infectious) particles. Bacterial and viral infections can induce the release of inflammatory cytokines and chemokines, including IL-8, IL-6 and TNF-α from airway epithelial cells. The major cause of epithelial barrier breakdown during lung inflammation is related to the damage of tight and adherens junctions 7-10. Due to the risk of development and the presence of antibiotic resistance, new avenues have to be explored to tackle (opportunistic) infections. The primary culture of bronchial epithelial cells is of particular importance, allowing the characterization of pathogenic infection in airway epithelium, advancing our knowledge of airway inflammation, and developing new intervention strategies. Because of the ethical and practical concerns, especially among vulnerable population groups, including children, it is highly challenging to obtain primary tissue for isolating primary bronchial epithelial cells (PBECs) from infants and children, which hinders progress in research. However, the bovine epithelium can be obtained on a regular basis in sufficient quantities from calves slaughtered for food co...

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Section: Discussionmentioning

confidence: 99%

Mannheimia haemolytica and lipopolysaccharide induce airway epithelial inflammatory responses in an extensively developed ex vivo calf model

Cai

Varasteh

Putten

et al. 2020

Sci Rep

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“…The model forms a heavily ciliated, mucus-producing cell layer with similar physiology to that of the native respiratory tissue. In tandem, we have developed a BBEC culture model which well represents the respiratory epithelium of cattle 26,27 . The sero-specific nature of severe cases of respiratory disease by M. haemolytica 11,38 , coupled with a suggested role for the polysaccharide capsule (on which the serotyping classification is based) in cell adhesion 17 , indicates that isolates may be specifically adapted to colonising the airway tissues of discrete hosts.…”

Section: Discussionmentioning

confidence: 99%

Differentiated ovine tracheal epithelial cells support the colonisation of pathogenic and non-pathogenic strains of Mannheimia haemolytica

O’Boyle

Berry

Davies

2020

Sci Rep

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Mannheimia haemolytica is the primary bacterial species associated with respiratory disease of ruminants. A lack of cost-effective, reproducible models for the study of M. haemolytica pathogenesis has hampered efforts to better understand the molecular interactions governing disease progression. We employed a highly optimised ovine tracheal epithelial cell model to assess the colonisation of various pathogenic and non-pathogenic M. haemolytica isolates of bovine and ovine origin. Comparison of single representative pathogenic and non-pathogenic ovine isolates over ten time-points by enumeration of tissue-associated bacteria, histology, immunofluorescence microscopy and scanning electron microscopy revealed temporal differences in adhesion, proliferation, bacterial cell physiology and host cell responses. Comparison of eight isolates of bovine and ovine origin at three key time-points (2 h, 48 h and 72 h), revealed that colonisation was not strictly pathogen or serotype specific, with isolates of serotype A1, A2, A6 and A12 being capable of colonising the cell layer regardless of host species or disease status of the host. A trend towards increased proliferative capacity by pathogenic ovine isolates was observed. These results indicate that the host-specific nature of M. haemolytica infection may result at least partially from the colonisation-related processes of adhesion, invasion and proliferation at the epithelial interface.

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“…Bovine bronchial epithelial cells can be stimulated to differentiate into a more representative model of the in vivo microenvironment through exposure to the atmosphere [36, 37, 59]. The methodology for differentiation of bovine airway epithelial cells have been fully optimised [44] and the model well-characterised [45]. The differentiated BBEC model has been shown to replicate the hallmark defences of the respiratory tract, including active mucocilary clearance.…”

Section: Discussionmentioning

confidence: 99%

“…We have previously investigated the growth conditions required for optimal growth and differentiation of bovine bronchial epithelial cells at an ALI [44] and assessed the temporal differentiation of these cells to identify an optimum window suitable for infection studies [45]. The aim of the present study was to investigate the interactions of a panel of M. haemolytica isolates, representing virulent and commensal strains recovered from both cattle and sheep, with differentiated bovine bronchial epithelial cells grown at an ALI.…”

Section: Introductionmentioning

confidence: 99%

Interactions of pathogenic and commensal strains of Mannheimia haemolytica with differentiated bovine airway epithelial cells grown at an air-liquid interface

Cozens

Sutherland

Lauder

et al. 2017

Preprint

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18Mannheimia haemolytica serotype A2 is a common commensal species present in the 19 nasopharynx of healthy cattle. However, prior to the onset of bovine pneumonic 20 pasteurellosis, there is sudden increase in M. haemolytica serotype A1 within the upper 21 respiratory tract. The events during this selective proliferation of serotype A1 strains are 22 [19, 20] and can also be distinguished by differences in their outer membrane protein (OMP) 58 profiles [21], lipopolysaccharide types [21, 22] and nucleotide sequence variation in various 59 virulence-associated genes including lktA [23], ompA [24], tbpA and tbpB [25], plpE [26] as 60 well as a number of other genes [20]. The upper respiratory tract of healthy cattle is 61predominantly colonized by serotype A2 strains but, for reasons that are not clear (but 62 probably related to stress and/or viral infection), a transition occurs within this 63 4 microenvironment which leads to a sudden explosive proliferation in the number of serotype 64 A1/A6 bacteria present and subsequent colonization [1, 3]. This sudden and selective 65 explosion in the A1/A6 population within the upper respiratory tract leads to the inhalation of 66 bacteria-containing aerosol droplets into the trachea and lungs and the onset of pneumonic 67 pasteurellosis [27]. Crucially, the specific bacterial and host factors responsible for the 68 sudden shift from commensal serotype A2 to pathogenic serotype A1/A6 populations within 69 the upper respiratory tract are not clear. 70The leukotoxin (LktA) of M. haemolytica plays a central role in the pathogenesis of 71 pneumonic pasteurellosis and significant attention has been given to understanding the 72 molecular mechanisms associated with LktA activity within the lung [1, 3, 28, 29]. In 73 contrast, there has been far less focus on the interactions of M. haemolytica with respiratory 74 airway epithelial cells and events that might account for the sudden proliferation of serotype 75 A1/A6 bacteria within the upper respiratory tract. A contributing factor to our poor 76 understanding of early host-pathogen interactions associated with pneumonic pasteurellosis, 77 and indeed BRD in general, is the lack of physiologically-relevant and reproducible 78 methodologies with which to study the intricate molecular and immunological interactions 79 between pathogens and host. Traditionally, submerged, two-dimensional cultures of a single 80 cell type have been used to investigate interactions of M. haemolytic and other BRD 81 pathogens within the bovine respiratory tract [30-32] but these have numerous limitations: 82 they do not reflect the multicellular complexity of the parental tissue in vivo, they lack its 83 three-dimensional (3-D) architecture, and the physiological conditions are not representative 84 of those found within the respiratory tract. However, these characteristics that are lacking in 85 submerged cultures can be recapitulated using differentiated airway epithelial cells (AECs) 86 grown at an air-liquid interface (ALI) and, in recent years, s...

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Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cited by 24 publications

References 92 publications

Mannheimia haemolytica and lipopolysaccharide induce airway epithelial inflammatory responses in an extensively developed ex vivo calf model

Mannheimia haemolytica and lipopolysaccharide induce airway epithelial inflammatory responses in an extensively developed ex vivo calf model

Differentiated ovine tracheal epithelial cells support the colonisation of pathogenic and non-pathogenic strains of Mannheimia haemolytica

Interactions of pathogenic and commensal strains of Mannheimia haemolytica with differentiated bovine airway epithelial cells grown at an air-liquid interface

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