Abstract
Legionnaires’ disease is a severe pneumonia caused by inhalation of Legionella pneumophila. Although powerful infection models ranging from monocellular host systems to mammals were developed, numerous intra- and extracellular interactions of L. pneumophila factors with human lung tissue structures remain unknown. Therefore, we developed and applied a novel infection model for Legionnaires’ disease comprising living human lung tissue explants (HLTEs). This model allows analyzing Legionella infections at a unique level of complexity and narrows the gap between current infection models and postmortem histopathology analyses of infected patients. Here we describe the infection of tumor-free pulmonary tissue samples from patients undergoing lobe- or pneumectomy because of lung cancer. The method comprises bacterial cultivation, preparation of HLTEs, and infection of HLTEs. The infected tissue samples allow to characterize tissue damage, bacterial localization, dissemination and growth kinetics, and the host’s molecular response.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Grivel JC, Margolis L (2009) Use of human tissue explants to study human infectious agents. Nat Protoc 4(2):256–269. https://doi.org/10.1038/nprot.2008.245
Abbot EL, Smith WD, Siou GP et al (2007) Pili mediate specific adhesion of Streptococcus pyogenes to human tonsil and skin. Cell Microbiol 9(7):1822–1833. https://doi.org/10.1111/j.1462-5822.2007.00918.x
Haque A, Bowe F, Fitzhenry RJ et al (2004) Early interactions of Salmonella enterica serovar typhimurium with human small intestinal epithelial explants. Gut 53(10):1424–1430
Greenhead P, Hayes P, Watts PS et al (2000) Parameters of human immunodeficiency virus infection of human cervical tissue and inhibition by vaginal virucides. J Virol 74(12):5577–5586. https://doi.org/10.1128/JVI.74.12.5577-5586.20
Fischetti L, Barry SM, Hope TJ et al (2009) HIV-1 infection of human penile explant tissue and protection by candidate microbicides. AIDS 23(3):319–328. https://doi.org/10.1097/QAD.0b013e328321b778
Nash TW, Libby DM, Horwitz MA (1984) Interaction between the legionnaires’ disease bacterium (Legionella pneumophila) and human alveolar macrophages. Influence of antibody, lymphokines, and hydrocortisone. J Clin Invest 74(3):771–782. https://doi.org/10.1172/JCI111493
Hambletoan P, Baskerville RB, Fitzgeorge B et al (1982) Pathological and biochemical features of Legionella pneumophila infection in guinea-pigs. J Med Microbiol 15:317–326. https://doi.org/10.1099/00222615-15-3-317
Brieland J, Freeman P, Kunkel R et al (1994) Replicative Legionella pneumophila lung infection in intratracheally inoculated A/J mice. A murine model of human Legionnaires’ disease. Am J Pathol 145(6):1537–1546
Rupp J, Droemann D, Goldmann T et al (2004) Alveolar epithelial cells type II are major target cells for C. pneumoniae in chronic but not in acute respiratory infection. FEMS Immunol Med Microbiol 41(3):197–203
Xu F, Droemann D, Rupp J et al (2008) Modulation of the inflammatory response to Streptococcus pneumoniae in a model of acute lung tissue infection. Am J Respir Cell Mol Biol 39(5):522–529. https://doi.org/10.1165/rcmb.2007-0328OC
Droemann D, Rupp J, Rohmann K et al (2010) The TGF-beta-Pseudoreceptor BAMBI is strongly expressed in COPD lungs and regulated by nontypeable Haemophilus influenzae. Respir Res 11:67. https://doi.org/10.1186/1465-9921-11-67
Szymanski KV, Toennies M, Becher A et al (2012) Streptococcus pneumoniae-induced regulation of cyclooxygenase-2 in human lung tissue. Eur Respir J 40(6):1458–1467. https://doi.org/10.1183/09031936.00186911
Hoppe J, Ünal CM, Thiem S et al (2017) PilY1 promotes Legionella pneumophila infection of human lung tissue explants and contributes to bacterial adhesion, host cell invasion, and twitching motility. Front Cell Infect Microbiol 7:63. https://doi.org/10.3389/fcimb.2017.00063
Jäger J, Marwitz S, Tiefenau J et al (2014) Human lung tissue explants reveal novel interactions during Legionella pneumophila infections. Infect Immun 82(1):275–285. https://doi.org/10.1128/IAI.00703-13
Shevchuk O, Abidi N, Klawonn F et al (2014) HOPE-fixation of lung tissue allows retrospective proteome and phosphoproteome studies. J Proteome Res 13(11):5230–5239. https://doi.org/10.1021/pr500096a
Bergmann S, Steinert M (2015) From single cells to engineered and explanted tissues: new perspectives in bacterial infection biology. In: Jeon KW (ed) International review of cell and molecular biology, vol 319. Academic Press, Cambridge, MA, pp 1–44
Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft (DFG STE 838/8-1).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Scheithauer, L., Steinert, M. (2019). Infection of Human Lung Tissue Explants (HLTEs) with Legionella pneumophila. In: Buchrieser, C., Hilbi, H. (eds) Legionella. Methods in Molecular Biology, vol 1921. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9048-1_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9048-1_21
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9047-4
Online ISBN: 978-1-4939-9048-1
eBook Packages: Springer Protocols