Asthma in fresh human lung tissue

Asthma in fresh human lung tissue

Asthma is one of the most common lung diseases worldwide. Main features of the disease can be displayed ex vivo by using fresh lung tissue, so-called precision-cut lung slices (PCLS). PCLS contain epithelial cells, fibroblasts, smooth muscle cells, nerve fibers, and even immune cells such as antigen-presenting cells and T-cells. The tissue is fully viable. Cells in the tissue interact with each other, thereby reflecting the highly specialized function of the lung.

We use lung tissue of laboratory animals and human donors. The tissue is exposed ex vivo to proteins known to be important in the pathophysiology of asthma, such as IL-13. The tissue can also be passively sensitized to commonly used allergens such as ovalbumin or house dust mite (HDM). The tissue is subsequently examined for immune responses, changes in cellular phenotype, respiratory toxicity, airway constriction and dilation, and vasoconstriction and dilation. Features of asthma can thus be investigated –using tissue of different species including human. We found the tissue response to be highly comparable with the in-vivo response, and it can be used for prediction of organ responses.

Endpoints/outcome parameters

  • Early airway response
  • Airway hyperresponsiveness
  • Proinflammatory responses

 

Readout parameters

  • Tissue viability: assessed by LDH assay, WST-1 assay, calcein AM/EthD-1 staining
  • Airway constriction by videomicroscopy: early airway response (EAR) – airway response to allergen exposure; airway hyperresponsiveness (AHR) – airway response to methacholine exposure
  • Proinflammatory responses of lung tissue: cytokine levels by ELISA or MSD, protein expression by Western blot
  • Histology imaging: conventional stainings, histopathology, immunohistochemistry, and scoring
  • Lung tissue analysis: RNA isolation for gene expression analysis

Publications

  1. Switalla S, Lauenstein L, Prenzler F, Knothe S, Förster C, Fieguth HG, Pfennig O, Schaummann F, Martin C, Guzman CA, Ebensen T, Müller M, Hohlfeld JM, Krug N, Braun A, Sewald K. Natural innate cytokine response to immunomodulators and adjuvants in human precision-cut lung slices. Toxicol Appl Pharmacol 246 (2010): 107-115.
  2. Switalla S, Knebel J, Ritter D, Krug N, Braun A, Sewald K. Effects of acute in vitro exposure of murine precision-cut lung slices to gaseous nitrogen dioxide and ozone in an air-liquid interface (ALI) culture. Toxicol Lett 196 (2010): 117-124.
  3. Seehase S, Schlepütz M, Switalla S, Mätz-Rensing K, Kaup FJ, Zöller M, Schlumbohm C, Fuchs E, Lauenstein HD, Winkler C, Kuehl AR, Uhlig S, Braun A, Sewald K, Martin C. Bronchoconstriction in nonhuman primates: a species comparison. J Appl Physiol 111 (2011): 791-798.
  4. Seehase S, Lauenstein HD, Schlumbohm C, Switalla S, Neuhaus V, Förster C, Fuchs E, Kaup FJ, Zöller M, Braun A, Sewald K, Knauf S. LPS-induced lung inflammation in marmoset monkeys – an acute model for anti-inflammatory drug testing. PLoS ONE August 28, 2012, doi: 10.1371/journal.pone.0043709.
  5. Lauenstein L, Switalla S, Prenzler F, Seehase S, Pfennig O, Förster C, Fieguth H, Braun A, Sewald K. Assessment of immunotoxicity induced by chemicals in human precision-cut lung slices (PCLS). Toxicol In Vitro 28 (2014): 588-599.
  6. Hess A, Wang-Lauenstein L, Braun A, Kolle SN, Landsiedel R, Liebsch M, Ma-Hock L, Pirow R, Schneider X, Steinfath M, Vogel S, Martin C, Sewald K. Prevalidation of the ex-vivo model PCLS for prediction of respiratory toxicity, Toxicol In Vitro 32 (2016): 347-61.