Continuous mucociliary transport by primary human airway epithelial cells in vitro

Mucociliary clearance (MCC) is an important innate defense mechanism that continuously removes inhaled pathogens and particulates from the airways. Normal MCC is essential for maintaining a healthy respiratory system, and impaired MCC is a feature of many airway diseases, including both genetic (cystic fibrosis, primary ciliary dyskinesia) and acquired (chronic obstructive pulmonary disease, bronchiectasis) disorders. Research into the fundamental processes controlling MCC, therefore, has direct clinical application, but has been limited in part due to the difficulty of studying this complex multicomponent system in vitro. In this study, we have characterized a novel method that allows human airway epithelial cells to differentiate into a mucociliary epithelium that transports mucus in a continuous circular track. The mucociliary transport device allows the measurement and manipulation of all features of mucociliary transport in a controlled in vitro system. In this initial study, the effect of ciliary beat frequency and mucus concentration on the speed of mucociliary transport was investigated.

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417: Cyclosporine does not penetrate, but accumulates at high concentrations within, differentiated human airway epithelial cells in vitro: Possible implications for the use of inhaled cyclosporine

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2006.11.439 ◽

2007 ◽

Vol 26 (2) ◽

pp. S210

Author(s):

R. Aris ◽

P. McNeillie ◽

O. Olusesi ◽

S. Thomas ◽

M. Paine ◽

...

Keyword(s):

Epithelial Cells ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Human Airway ◽

High Concentrations ◽

Human Airway Epithelial Cells

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Pseudomonaspyocyanine alters calcium signaling in human airway epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1998.274.6.l893 ◽

1998 ◽

Vol 274 (6) ◽

pp. L893-L900 ◽

Cited By ~ 19

Author(s):

Gerene M. Denning ◽

Michelle A. Railsback ◽

George T. Rasmussen ◽

Charles D. Cox ◽

Bradley E. Britigan

Keyword(s):

Epithelial Cells ◽

Lung Disease ◽

Cell Function ◽

Oxidant Stress ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Human Airway ◽

Human Airway Epithelial Cells

Pseudomonas aeruginosa, an opportunistic human pathogen, causes both acute and chronic lung disease. P. aeruginosa exerts many of its pathophysiological effects by secreting virulence factors, including pyocyanine, a redox-active compound that increases intracellular oxidant stress. Because oxidant stress has been shown to affect cytosolic Ca2+concentration ([Ca2+]c) in other cell types, we studied the effect of pyocyanine on [Ca2+]cin human airway epithelial cells (A549 and HBE). At lower concentrations, pyocyanine inhibits inositol 1,4,5-trisphosphate formation and [Ca2+]cincreases in response to G protein-coupled receptor agonists. Conversely, at higher concentrations, pyocyanine itself increases [Ca2+]c. The pyocyanine-dependent [Ca2+]cincrease appears to be oxidant dependent and to result from increased inositol trisphosphate and release of Ca2+from intracellular stores. Ca2+plays a central role in epithelial cell function, including regulation of ion transport, mucus secretion, and ciliary beat frequency. By disrupting Ca2+homeostasis, pyocyanine could interfere with these critical functions and contribute to the pathophysiological effects observed in Pseudomonas-associated lung disease.

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Ozone-induced release of cytokines and fibronectin by alveolar macrophages and airway epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1994.266.6.l612 ◽

1994 ◽

Vol 266 (6) ◽

pp. L612-L619 ◽

Cited By ~ 28

Author(s):

R. B. Devlin ◽

K. P. McKinnon ◽

T. Noah ◽

S. Becker ◽

H. S. Koren

Keyword(s):

Epithelial Cells ◽

Cell Line ◽

Alveolar Macrophages ◽

Airway Epithelial Cells ◽

Airway Hyperreactivity ◽

Ozone Exposure ◽

Airway Epithelial ◽

Human Airway ◽

Human Airway Epithelial Cells

Acute exposure of animals and humans to ozone results in decrements in lung function, development of airway hyperreactivity, inflammation, edema, damage to pulmonary cells, and production of several compounds with tissue damaging, fibrinogenic or fibrotic potential. The contribution of airway epithelial cells and alveolar macrophages to these processes is unclear. In this study we have directly exposed human alveolar macrophages and human airway epithelial cells to ozone in vitro and measured the cytotoxic effects of ozone, as well as the production of the inflammatory cytokines interleukin-6 (IL-6) and interleukin-8 (IL-8), and fibronectin, all of which are substantially elevated in the bronchoalveolar lavage fluid of humans exposed to ozone. Cells were grown on rigid, collagen-impregnated filter supports, and the interaction of cells with ozone facilitated by exposing them to the gas with medium below the support but no medium on top of the cells. The results show that, although macrophages are much more sensitive to ozone than epithelial cells, they do not produce increased amounts of IL-6, IL-8, or fibronectin following ozone exposure. In contrast, epithelial cells produce substantially more of all three proteins following ozone exposure, and both IL-6 and fibronectin are secreted vectorially. An immortalized human airway epithelial cell line (BEAS 2B) was used in these experiments since human airway epithelial cells are infrequently available for in vitro studies. Data from this study extend previous findings which suggest that the BEAS cell line is a useful model to study the interaction between airway epithelial cells and environmental toxicants.

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Mechanisms of innate immunity involvement in airway disease exacerbations: Experiments with in vitro models of human airway epithelial cells (EpiAirway™) and epithelial cell/fibroblast co-cultures (EpiAirway-FT™)

Toxicology Letters ◽

10.1016/j.toxlet.2009.06.692 ◽

2009 ◽

Vol 189 ◽

pp. S168

Author(s):

Patrick Hayden ◽

Helena Kandarova ◽

George Jackson ◽

Alexander Armento ◽

Jennifer Bolmarcich ◽

...

Keyword(s):

Innate Immunity ◽

Epithelial Cell ◽

Epithelial Cells ◽

Airway Disease ◽

Airway Epithelial Cells ◽

In Vitro Models ◽

Airway Epithelial ◽

Human Airway ◽

Human Airway Epithelial Cells

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Alpha 1-adrenergic signaling in human airway epithelial cells involves inositol lipid and phosphate metabolism

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1992.262.2.l183 ◽

1992 ◽

Vol 262 (2) ◽

pp. L183-L191 ◽

Cited By ~ 1

Author(s):

C. M. Liedtke

Keyword(s):

Epithelial Cells ◽

Inositol Phosphates ◽

Inositol Phosphate ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Nasal Epithelial Cells ◽

Human Airway ◽

Adrenergic Antagonist ◽

Human Airway Epithelial Cells

A role for phospholipase C (PLC) hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) as a mechanism of alpha 1-adrenergic signal transduction in human airway epithelial cells (AEC) was investigated in isolated normal tracheal and cystic fibrosis (CF) nasal epithelial cells grown in in vitro culture and prelabeled with 3 muCi myo-[3H]inositol/ml for 72 h. Breakdown of polyphosphoinositides was measured using thin-layer chromatography to detect phosphatidylinositol, phosphatidylinositol 4-phosphate (PIP), and PIP2. Inositol phosphates were separated by ion-exchange column chromatography. In normal AEC, the addition of the endogenous catecholamine l-epinephrine produced a rapid, transient accumulation of inositol 1,4,5-trisphosphate (IP3) and inositol 1,4-bisphosphate (IP2) and breakdown of PIP and PIP2. IP3 increased 1.7-fold and IP2 1.6-fold after 20 and 40 s, respectively. A maximal decrease of 35% PIP2 and 30% PIP is observed after 20 and 40 s, respectively. The effects of l-epinephrine were not blocked by the beta-adrenergic antagonist dl-propranolol but were mimicked by the alpha 1-adrenergic agonist methoxamine. Prazosin, an alpha 1-adrenergic antagonist, and pertussis toxin (PTX) blocked the effects of l-epinephrine and methoxamine. Addition of l-epinephrine and methoxamine to CF nasal epithelial cells also induced prazosin-sensitive polyphosphoinositide breakdown and inositol phosphate accumulation. A 2.2-fold accumulation of IP3 was observed after 10 s and 2.0-fold increase in IP2 after 20 s. Maximal decreases of 32% PIP2 and 23% PIP levels were observed after 20-s incubation with l-epinephrine. PTX reduced the effects of l-epinephrine and significantly blocked the effects of methoxamine.(ABSTRACT TRUNCATED AT 250 WORDS)

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