ToxSci Advance Access originally published online on September 4, 2008
Toxicological Sciences 2008 106(2):444-453; doi:10.1093/toxsci/kfn178
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Ex Vivo Lung Function Measurements in Precision-Cut Lung Slices (PCLS) from Chemical Allergen–Sensitized Mice Represent a Suitable Alternative to In Vivo Studies
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* Department of Immunology, Allergology and Immunotoxicology, Fraunhofer Institute of Toxicology and Experimental Medicine, 30625 Hannover, Germany
Institute of Pharmacology and Toxicology, RWTH Aachen, Wendlingweg 2, 52074 Aachen, Germany and Division of Pulmonary Pharmacology, Research Center Borstel, 23845 Borstel, Germany
1 To whom correspondence should be addressed at Department of Immunology, Allergology and Immunotoxicology, Fraunhofer Institute of Toxicology and Experimental Medicine, Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany. Fax: +49-511-5350-155. E-mail: armin.braun{at}item.fraunhofer.de.
Received June 23, 2008; accepted August 18, 2008
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Abstract |
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A wide range of industrial chemicals can induce respiratory allergic reactions. Hence, there is an urgent need for methods identifying and characterizing the biological action of chemicals in the lung. Here, we present an easy, reliable alternative method to measure lung function changes ex vivo after exposure to chemical allergens and compare this to invasive in vivo measurements after sensitization with the industrial chemicals trimellitic anhydride (TMA) and 2,4-dinitrochlorobenzene (DNCB). Female BALB/c mice were sensitized epicutaneously with the respiratory allergen TMA and the contact sensitizer DNCB. The early allergic response to TMA and DNCB was registered in vivo and ex vivo on day 21 after inhalational challenge with dry standardized aerosols or after exposure of precision-cut lung slices (PCLS) to dissolved allergen. Airway hyperresponsiveness (AHR) to increasing doses of methacholine (MCh) was measured on the next day in vivo and ex vivo. Bronchoalveolar lavage (BAL) was performed for immunological characterization of local inflammation. TMA-sensitized mice showed AHR to MCh in vivo (ED50: 0.06 µg MCh vs. 0.21 µg MCh in controls) and in PCLS (EC50: 0.24 µM MCh vs. 0.4 µM MCh). TMA-treated animals showed increased numbers of eosinophils (12.8·104 vs. 0.7·104) and elevated eotaxin-2 concentrations (994 pg/ml vs. 167 pg/ml) in BAL fluid 24 h after allergen challenge. In contrast, none of these parameters differed after sensitization with DNCB. The present study suggests that the effects of low molecular weight allergens, like TMA and DNCB, on ex vivo lung functions tested in PCLS reflect the in vivo situation.
Key Words: lung function; bronchoconstriction; PCLS; TMA; DNCB; methacholine.
2 These authors contributed equally to the study.