Fraunhofer Institute for Toxicology and Experimental Medicine
Fraunhofer Institute for Toxicology and Experimental Medicine

Chronic obstructive pulmonary disease (COPD)

The common chronic obstructive pulmonary disease (COPD) is an increasingly important cause of morbidity and mortality. More than 200 million people worldwide are affected by COPD, making it the fourth leading cause of death. COPD is characterized by progressive destruction of the lung parenchyma, resulting in the development of emphysema, increased mucus production, and chronic airway inflammation. It is a highly heterogeneous disease. Cigarette smoke has been shown to be the most important risk factor for the development of COPD; but also burning of biomass fuels and occupational exposure are known to be COPD risk factors.

At Fraunhofer ITEM, we use primary cells, cell lines, fresh lung tissue, and animal models to induce features of COPD. Lipopolysaccharides (LPS), cigarette smoke, and cigarette smoke condensate are applied as agents. LPS-induced inflammation mimics primarily an acute and subacute inflammatory process in the lung. We have models of LPS-induced inflammation in laboratory animals including non-human primates, in fresh lung tissue and cells. Cells and tissue can also be exposed to cigarette smoke condensate and cigarette smoke. This is done primarily at the air-liquid interface. In particular the exposure of cells and tissue at the air-liquid interface is considered to closely reflect the human situation.

Acute and subacute respiratory inflammation

Expand all Close all

  • Inflammation is an essential component of many respiratory diseases, including pneumonia, asthma and chronic obstructive pulmonary disease (COPD). Main features of respiratory inflammation can be displayed in animal models, including non-human primates. Marmosets in particular are new world monkeys with a very high level of homology and physiological similarity to humans.

    In our model, acute inflammation is induced by single or repeated exposure to endotoxin (lipopolysaccharide, LPS). This exposure treatment has been shown to reproduce hallmarks of human inflammation, with high neutrophil counts and sensitivity to glucocorticoid treatment. It is a robust, easy, and cost-efficient model that can be used especially for efficacy testing of anti-inflammatory drugs. We have excellent expertise in inhalation of substances and lung function measurements.

     

    Species:

    • Mouse 
    • Rat
    • Marmoset monkey

     

    Endpoints/outcome parameters:

    • Acute airway inflammation

     

    Readout parameters:

    • In-life assessment of lung function: airway response to methacholine exposure
    • Bronchoalveolar lavage: total and differential cell counts, cytokine levels by ELISA or MSD 
    • Histology imaging: conventional stainings, histopathology, immunohistochemistry, and pathology scoring 

     

    Publications:

    1. Curths C, Wichmann J, Dunker S, Windt H, Hoymann HG, Lauenstein HD, Hohlfeld J, Becker T, Kaup FJ, Braun A, and Knauf S. Airway hyper-responsiveness in lipopolysaccharide-challenged common marmosets (Callithrix jacchus). Clin Sci (Lond). 126 (2014), No. 2: 155-162.
    2. Seehase S, Schleputz M, Switalla S, Matz-Rensing K, Kaup FJ, Zoller M, Schlumbohm C, Fuchs E, Lauenstein HD, Winkler C, Kuehl AR, Uhlig S, Braun A, Sewald K, and Martin C. Bronchoconstriction in nonhuman primates: a species comparison. J Appl Physiol. 111 (2011), No. 3: 791-798.
    3. Seehase S, Lauenstein HD, Schlumbohm C, Switalla S, Neuhaus V, Forster C, Fieguth HG, Pfennig O, Fuchs E, Kaup FJ, Bleyer M, Hohlfeld JM, Braun A, Sewald K, and Knauf S. LPS-induced lung inflammation in marmoset monkeys - an acute model for anti-inflammatory drug testing. PLoS One. 7 (2012), No. 8: e43709.

Fresh human lung tissue and cells

Expand all Close all

  • The common chronic obstructive pulmonary disease (COPD) is characterized by progressive destruction of the lung parenchyma. More than 200 million people worldwide are affected by COPD. Cigarette smoke has been shown to be majorly responsible for COPD pathogenesis. Main features of COPD 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 cigarette smoke and cigarette smoke condensate using an air-liquid culture. The PCLS are subsequently examined for immune responses, changes in cellular phenotype, and respiratory toxicity. Features of COPD 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.

     

    Species:

    • Mouse 
    • Rat 
    • Monkey
    • Human

     

    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.