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Precision cut lung slices as tool for assessment of allergens

: Switalla, S.; Sewald, K.; Henjakovic, M.; Veres, T.; Krug, N.; Braun, A.


44th Congress of the European Societies of Toxicology, EUROTOX 2007. Abstracts : Amsterdam, The Netherlands, 7 - 10 October 2007
Amsterdam: Elsevier, 2007 (Toxicology letters 172.2007, Supplement 1)
ISSN: 0378-4274
European Societies of Toxicology (Congress) <44, 2007, Amsterdam>
Conference Paper, Journal Article
Fraunhofer ITEM ()
precision cut lung slice; assessment; allergen

Studies on precision cut lung slices (PCLS) are part of the European Union project SENS-IT-IV. Aim of this project is the development and establishment of in vitro techniques for assessment of allergenic potential of inhaled substances without animal testing.
Mouse PCLS were prepared and treated with subtoxic concentrations of respiratory (ammonium-hexachloroplatinate, AHCP) and contact (cinnamaldehyde) allergens in comparison to negative control (salicylic acid). Supernatants were collected, PCLS were lysed and extrinsic and intrinsic cytokine profiles were analysed by ELISA and cytokine array. In addition lung slices were stained with calcein and anti-ICAM-1 for confocal microscopy.
Cytokine levels showed significant increases after treatment with AHCP compared to tissue control. Interleukine (IL)-10 was increased up to 2.2-fold, TNF-alpha and IL-1alpha up to 1.6-fold and MIP-1beta up to 0.7-fold. Cytokines like eotaxin, G-CSF and IL-12 showed a 0.75-fold increase. Stimulation with cinnamaldehyde resulted in a significant decrease in MIP-1beta up to 0.5-fold, but showed no significant changes in expression of other cytokines. Salicylic acid showed compared to AHCP an increase of TNF-alpha-expression, but e.g. no increase of IL-1alpha. Staining of epithelial cells showed an upregulation of ICAM-1 in response to activation.
Our experiments showed different expression patterns of cytokines for representative respiratory and contact allergens and indicate that PCLS is a usable ex vivo model for the assessment of new substances.