An in vitro coculture system for the detection of sensitization following aerosol exposure

February 20, 2019


Aline Chary1,2, Tommaso Serchi1, Elisa Moschini1, Jennifer Hennen2, Sébastien Cambier1, Janine Ezendam3, Brunhilde Blömeke2 and Arno C. Gutleb1
1Department of Environmental Research and Innovation, Luxembourg Institute of Science and Technology, Luxembourg; 
2Department of Environmental Toxicology, University Trier, Germany; 
3Centre for Health Protection, National Institute for Public Health and the Environment, The Netherlands


The Vitrocell® Cloud-6 system was used for the exposure of a coculture in vitro model to test compounds and vehicle controls. The 3D in vitro model was cultured at the air liquid interface consists of A549, EA.hy926, PMA-differentiated THP-1 and non-differentiated THP-1 cells, that were exposed to nebulized chemical respiratory sensitizers. The results are shown in flow cytometry measurements, cytokine release and gene expression.


The aim of the study was to develop an in vitro model that mimics the alveolar-capillary barrier and that allows assessment of the respiratory sensitizing potential of respiratory sensitizers. The 3D in vitro model cultured at the air liquid interface consists of alveolar type II epithelial cells (A549), endothelial cells (EA.hy926), macrophage-like cells (PMA-differentiated THP-1) and dendritic-like cells (non-differentiated THP-1). This alveolar model was exposed apically to nebulized chemical respiratory sensitizers (Phthalic Anhydride (PA) and TriMellitic Anhydride (TMA)) or irritants (Methyl Salicylate (MeSa) and Acrolein (Acr)) at concentrations inducing at maximum 25% of cytotoxicity. The exposure to respiratory sensitizers induced dendritic cells activation and a specific cytokine release pattern, while the irritants did not. In addition, the cell surface marker OX40L was determined for dendritic like cells activation to identify high molecular weight allergens. With this in vitro model we can postulate a set of promising markers based on the studied compounds that allow the discrimination of chemical respiratory sensitizers from irritants.

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