10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a4662

Assessment of lung toxicity induced by chemical allergens in human precision-cut lung slices

Abstract

Introduction: A number of low-molecular weight (LMW) chemicals at workplaces are involved in the development of occupational asthma. There are no currently accepted and validated test methods to identify chemicals with the potential to cause respiratory sensitization. Risk assessment is normally performed in animal experiments, however, in the context of REACH and its principle of 3Rs there is an increasing public demand for alternative methods. Human precision cut lung slices (PCLS) is an ex vivo approach in which all relevant cell types of lung tissue are present in their natural position. The aim of the study was to analyse chemical-induced inflammation and irritation by assessing a variety of immunotoxic endpoints in human PCLS. Methods: Human PCLS were prepared from tumor-free tissue from resected lung lobes of cancer patients. PCLS were incubated with 20 substances in DMEM under standard submerged cell culture conditions. PCLS without test substances were incubated as controls. After 24 hours incubation, induced toxicity was assessed by vitality staining and determination of enzymatic activity using WST-1 assay. EC25 with respect to vehicle controls were calculated for all chemicals. Cytokine contents were detected with MSD technology and ELISA. Results: Individual EC25 values correlated significantly with data published for in vitro approaches with THP-1 and NCTC cell lines (r2 = 0.87 and 0.83, respectively) (Mitjans et al. 386-95;Corsini et al. 789-96). Furthermore EC25 of human PCLS correlated with LD50 data published for in vivo rat inhalation toxicity with r of 0.53 (p value of 0.08). Subtoxic concentrations were used to investigate cytokine pattern for the differentiation between allergen and non-allergen. Respiratory allergens trimellitic anhydride and ammonium hexachloroplatinate increased the expression of the cytokines TNF? and IL-1? significantly while contact allergens (cinnamaldehyde, 2-bromo-2-glutaronitrile) and controls (phenol, SLS) failed to induce these cytokines to the same extent. Conclusion: The toxicity of chemicals in human PCLS resembles the in vitro situation very closely and also correlates with LD50 values from in vivo studies. The pro-inflammatory effects could be shown for some respiratory allergens. However, some respiratory allergens like glutaraldehyde did not induce cytokine due to the chemical properties. Altogether, PCLS could be regarded as an ex vivo immunotoxicity model, displaying chemical-induced inflammation and irritation on all relevant cell types of lung tissue

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oai:fraunhofer.de:N-226395Last time updated on 11/15/2016

This paper was published in Fraunhofer-ePrints.

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