Innate immune response in human lung tissue

Inflammation is an essential component of many respiratory diseases, including pneumonia, asthma and chronic obstructive pulmonary disease (COPD). Main features of inflammation 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 mitogens such as endotoxin (lipopolysaccharide, LPS) or polyI:C. This exposure has been shown to reproduce hallmarks of human inflammation, with high release of cytokines and chemokines responsible for recruiting of neutrophils and monocytes. The tissue is subsequently examined for immune responses, changes in cellular phenotype, respiratory toxicity, airway constriction and dilation, and vasoconstriction and -dilation. Features of inflammation 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.

Endpoint/outcome parameter

  • Proinflammatory responses

Readout parameters

  • Tissue viability: assessed by LDH assay, WST-1 assay, calcein AM/EthD-1 staining
  • Airway constriction by videomicroscopy: 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 M, 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.
  7. Sewald K and Braun A. PCLS: From learning about ‘in vivo’ to reduction and replacement. In: Progress towards novel testing strategies for in vitro assessment of allergens. Roggen EL, Weltzien HU, Hermans H (eds.). Kerala/India: Research Signpost, 2011, pp. 13-34.
  8. Sewald K and Braun A. Precision-cut tissue slices in pharmacology and toxicology. Xenobiotica 43 (2013): 84-97.