Assessment of immunotoxicity induced by chemicals in human precision-cut lung slices (PCLS)

Abstract

Occupational asthma can be induced by a number of chemicals at the workplace. Risk assessment of potential sensitizers is mostly performed in animal experiments. With increasing public demand for alternative methods, human precision-cut lung slices (PCLS) have been developed as an ex vivo model. Human PCLS were exposed to increasing concentrations of 20 industrial chemicals including 4 respiratory allergens, 11 contact allergens, and 5 non-sensitizing irritants. Local respiratory irritation was characterized and expressed as 75% (EC25) and 50% (EC50) cell viability with respect to controls. Dose-response curves of all chemicals except for phenol were generated. Local respiratory inflammation was quantified by measuring the production of cytokines and chemokines. TNF-α and IL-1α were increased significantly in human PCLS after exposure to the respiratory sensitizers trimellitic anhydride (TMA) and ammonium hexachloroplatinate (HClPt) at subtoxic concentrations, while contact sensitizers and non-sensitizing irritants failed to induce the release of these cytokines to the same extent. Interestingly, significant increases in T(H)1/T(H)2 cytokines could be detected only after exposure to HClPt at a subtoxic concentration. In conclusion, allergen-induced cytokines were observed but not considered as biomarkers for the differentiation between respiratory and contact sensitizers. Our preliminary results show an ex vivo model which might be used for prediction of chemical-induced toxicity, but is due to its complex three-dimensional structure not applicable for a simple screening of functional and behavior changes of certain cell populations such as dendritic cells and T-cells in response to allergens.

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

“…They form an attractive alternative to human epithelial cell cultures, since they contain all cell types in their in situ environment. Lauenstein et al reported on the use of human PCLS for chemical-induced toxicity, but concluded that it was not suitable for simple screening (Lauenstein et al, 2014). Use of this method is limited by the complex 3D structure of the model, the variable life span of the different cell types cultured, and the requirement for regular availability of fresh (healthy) human lung tissue.…”

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

“…Human precision-cut lung slices (PCLS), an ex vivo model consisting of all relevant cell types in their microanatomical environment (de Kanter et al, 2002), are well-established in pharmacological testing (Sturton et al, 2008), analysis of allergic immune responses (Wohlsen et al, 2003) and studies of xenobiotic metabolism (Lauenstein et al, 2014). There is also emerging literature concerning the use of ex vivo lung perfusion as a tool for experimental research (Nelson et al, 2014).…”

Evaluation of inflammatory and immune responses in long-term cultured human precision-cut lung slices

“…[1][2][3][4][5] Multiple studies have focused on assessing various characteristics of cultured PCLS, such as measurement of bronchoconstriction [6][7][8] and vascular responses 9,10 induced by various stimuli. In addition, PCLS have been used to evaluate allergy, 11 asthma, 6 toxicology, [12][13][14] and infectious disease responses. 15 The anatomical architecture of PCLS also allows for studying early inflammatory and immune responses that occur in the lung.…”