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Cytokine production after acute exposure of precision cut lung slices to NO2 and O3

: Switalla, S.; Ritter, D.; Krug, N.; Knebel, J.; Braun, A.; Sewald, K.


46th Congress of the European Societies of Toxicology 2009. Abstracts : Dresden, Germany, 13th - 16th September 2009; EUROTOX 2009
Amsterdam: Elsevier, 2009 (Toxicology letters 189.2009, Supplement 1)
ISSN: 0378-4274
European Societies of Toxicology (EUROTOX Congress) <46, 2009, Dresden>
Fraunhofer ITEM ()

Gaseous compounds like nitrogen dioxide and ozone display a major health risk. Consequence of inhalation can be associated with exacerbation of asthma, chronic pulmonary disease (COPD) or pneumonia. Precision cut lung slices (PCLS) represent an ex vivo technique which delivers the possibility to gain insight into chemical-induced effects in all cell types of the respiratory tract after air/lifted exposure to gaseous mixtures.
PCLS were prepared and exposed air/lifted to different concentrations of O3 and NO2. As negative control PCLS were exposed to clean air. Cytotoxicity was determined by WST-1 and live/dead staining for confocal microscopy. Chemical-induced inflammation was characterized by quantification of cytokine release by ELISA or Luminex technology.
WST-1 or live/dead staining showed that air/lifted cultivation and 1 h exposure to concentrations from 1 to 10 ppm NO2 or 3.5 to 8.5 ppm O3 did not induce cytotoxicity. NO2 induced cytotoxicity after exposure to a concentration of 70 ppm. Expression of IL-1alpha showed for NO2 a dose-dependent increase of up to 20% compared to clean air whereas RANTES decreased. Exposure to O3 resulted in dose-dependent and significant up regulation of proinflammatory cytokine IL-1alpha of up to 80 % whereas RANTES showed an increase of up to 800%. Cytokines like MCP-1 or IL-12 showed no changes for any gas compound.
These experiments pointed up that PCLS can be exposed air/lifted to gaseous compounds and show a dose-dependent cytotoxicity and cytokine production. This offers opportunities to investigate ex vivo immunotoxicological parameters for gaseous and particulate chemicals in a multifunctional test system.