Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

A method to enable the investigation of murine bronchial immune cells, their cytokines and mediators

Nature Protocols volume 2, pages 105–112 (2007)Cite this article

Abstract

Innovative therapies for severe lung diseases (such as allergic and chronic asthma, chronic obstructive pulmonary disease or any type of lung cancer) require a detailed understanding of the cellular and immune processes in the lung. This protocol details a method to obtain the immune cells of the bronchi as well as the cytokines and mediators produced by these cells for further investigation. The broncho-alveolar lavage fluid (BALF) is taken by injecting physiological solution through the tracheal tube into the murine airways and carefully regained by winding up the connected syringe. After centrifugation, the resulting BALF supernatant can be stored for detection of cytokines or other mediators by enzyme-linked immunosorbent assay or other methods; the resuspended cell pellet can also be used for flow cytometric analyses, to check cell viability and the level of apoptosis, as well as other applications. In addition, CD4+ T cells isolated from wild-type and genetically modified mice alone or along with other immunologically important cells such as T regulatory cells, which can be used to reconstitute immunodeficient mice, may be retrieved from the airways with this method. This protocol can be completed within 35 min.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Schematic drawing of the experimental retrieval of BALF.
Figure 2: FACS analysis of the BALF cells in wild-type and immunodeficient mice reconstituted with CD4+ T cells.

Similar content being viewed by others

References

  1. Doganci, A. et al. The IL-6R αchain controls lung CD4+CD25+ Treg development and function during allergic airway inflammation in vivo . J. Clin. Invest. 115, 313–325 (2005).

    Article  CAS  Google Scholar 

  2. Finotto, S. et al. Development of spontaneous airway changes consistent with human asthma in mice lacking T-bet. Science 295, 336–338 (2002).

    Article  CAS  Google Scholar 

  3. Finotto, S. et al. Treatment of allergic airway inflammation and hyperresponsiveness by antisense-induced local blockade of GATA-3 expression. J. Exp. Med. 193, 1247–1260 (2001).

    Article  CAS  Google Scholar 

  4. Finotto, S. et al. Local administration of antisense phosphorothioate oligonucleotides to the c-kit ligand, stem cell factor, suppresses airway inflammation and IL-4 production in a murine model of asthma. J. Allergy Clin. Immunol. 107, 279–286 (2001).

    Article  CAS  Google Scholar 

  5. Sauer, K.A., Scholtes, P., Kavwot, R. & Finotto, S. Isolation of CD4+ T Cells from murine lungs: a method to analyze ongoing immune-responses in the lung. Nat. Protocol in the press.

  6. Drazen, J.M. Leukotrienes in asthma. Adv. Exp. Med. Biol. 525, 1–5 (2003).

    Article  CAS  Google Scholar 

  7. Vargaftig, B.B. & Singer, M. Leukotrienes mediate murine bronchopulmonary hyperreactivity, inflammation, and part of mucosal metaplasia and tissue injury induced by recombinant murine interleukin-13. Am. J. Resp. Cell Mol. Biol. 28, 410–419 (2003).

    Article  CAS  Google Scholar 

  8. Jacobson, J.R. et al. Simvastatin attenuates vascular leak and inflammation in murine inflammatory lung injury. Am. J. Physiol. Lung Cell Mol. Physiol. 288, L1026–L1032 (2005).

    Article  CAS  Google Scholar 

  9. Ware, L.B. & Matthay, M.A. The acute respiratory distress syndrome. N. Engl. J. Med. 342, 1334–1349 (2000).

    Article  CAS  Google Scholar 

  10. Finotto, S. et al. Identification of epithelial cells in bronchoalveolar lavage. Hum. Exp. Toxicol. 12, 43–46 (1993).

    Article  CAS  Google Scholar 

  11. Finotto, S. et al. Analysis of mice carrying targeted mutations of the glucocorticoid receptor gene argue against an essential role of glucocorticoid signalling for generating adrenal chromaffin cells. Development 126, 2935–2944 (1999).

    CAS  PubMed  Google Scholar 

  12. Finotto, S. et al. Severe hepatic injury in interleukin 18 (IL-18) transgenic mice: a key role for IL-18 in regulating hepatocyte apoptosis in vivo . Gut 53, 392–400 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Christine Lux and Isabel Ernst for their technical support. This work was supported by an SFB 548 (projects B8 and Z3).

Author information

Author notes

  1. Joachim H Maxeiner and Roman Karwot: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cellular and Molecular Immunology of the Lung University of Mainz, Mainz, Germany

    Joachim H Maxeiner, Roman Karwot, Kerstin A Sauer & Susetta Finotto

  2. Asthma Core Facility, I Medical Clinic, Obere Zahlbacher Strasse 63, University of Mainz, Mainz, Germany

    Michael Hausding, Petra Scholtes & Susetta Finotto

Authors
  1. Joachim H Maxeiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roman Karwot
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Hausding
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kerstin A Sauer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Petra Scholtes
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Susetta Finotto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Susetta Finotto.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Maxeiner, J., Karwot, R., Hausding, M. et al. A method to enable the investigation of murine bronchial immune cells, their cytokines and mediators. Nat Protoc 2, 105–112 (2007). https://doi.org/10.1038/nprot.2007.8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2007.8

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing