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Isolation of CD4+ T cells from murine lungs: a method to analyze ongoing immune responses in the lung

Nature Protocols volume 1, pages 2870–2875 (2006)Cite this article

Abstract

The regulation of the cellular immune response in lung diseases is not yet fully understood. Isolating different subsets of immune cells directly from the lung is therefore an indispensable method of gaining detailed knowledge on the function of these cells in this organ. This protocol describes a method of isolating and magnetically labeling CD4+ lung T cells, which are then loaded and retained on the column while all other cells run through it (positive selection). The yield of this isolation is approximately 5 × 105 to 1.5 × 106 CD4+ cells from a murine lung. These cells can be further investigated by several methods such as flow cytometry, western blot analysis, RT-PCR, immunostaining and ELISA. In addition, lung CD4+ T cells alone or along with other immunologically important cells such as CD8+ T cells and T regulatory cells can be adoptively transferred into immune-deficient mice, and can influence important local parameters. This protocol can be completed in approximately 4 h 20 min.

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Figure 1: Schematic representation of the CD4+ lung T cell isolation.
Figure 2: FACS analysis of the total lung cells in wild-type mice.

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References

  1. Van Oosterhout, A.J. & Bloksma, N. Regulatory T-lymphocytes in asthma. Eur. Respir. J. 26, 918–932 (2005).

    Article  CAS  Google Scholar 

  2. Doganci, A., Sauer, K., Karwot, R. & Finotto, S. Pathological role of IL-6 in the experimental allergic bronchial asthma in mice. Clin. Rev. Allergy Immunol. 28, 257–270 (2005).

    Article  CAS  Google Scholar 

  3. Sinigaglia, F. & D'Ambrosio, D. Regulation of helper T cell differentiation and recruitment in airway inflammation. Am. J. Respir. Crit. Care Med. 162, S157–160 (2000).

    Article  CAS  Google Scholar 

  4. Neurath M.F., Finotto, S & Glimcher, L.H. The role of Th1/Th2 polarization in mucosal immunity. Nat. Med. 8, 567–73 (2002).

    Article  CAS  Google Scholar 

  5. Doganci, A. et al. The IL-6R alpha chain 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 

  6. Hausding, M. et al. A stage-specific functional role of the leucine zipper transcription factor c-Maf in lung Th2 cell differentiation. Eur. J. Immunol. 34, 3401–3412 (2004).

    Article  CAS  Google Scholar 

  7. Yoo, J.K. et al. IL-12 Provides proliferation and survival signals to murine CD4 T cells through phosphatidylinositol 3-kinase/Akt signaling pathway. J. Immunol. 169, 3637–3643 (2002).

    Article  CAS  Google Scholar 

  8. Yang, J. et al. Schistosoma japonicum egg antigens stimulate CD4 CD25 T cells and modulate airway inflammation in a murine model of asthma. Immunology, published online 17 October 2006 (doi: 10.1111/j.1365–2567.2006.02472.x).

  9. Quinn, K.M. et al. Inactivation of CD4+CD25+ regulatory T cells during early mycobacterial infection increases cytokine production but does not affect pathogen load. Immunol. Cell Biol. 84, 467–474 (2006).

    Article  CAS  Google Scholar 

  10. Finotto, S. et al. Asthmatic changes in mice lacking T-bet are mediated by IL-13. Int Immunol. 17, 993–1007 (2005).

    Article  CAS  Google Scholar 

  11. Jelley-Gibbs, D.M. et al. Unexpected prolonged presentation of influenza antigens promotes CD4 T cell memory generation. J. Exp. Med. 202, 697–706 (2005).

    Article  CAS  Google Scholar 

  12. Sakoda, Y. et al. Donor-derived thymic-dependent T cells cause chronic graft-versus-host disease. Blood, published online 10 October 2006 (doi: 10.1182/blood-2006-08-042853).

  13. 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 

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Acknowledgements

This work is supported by German grant SFB 548-B8-Z3. The authors thank Professor Jürgen Knop for his support to the Asthma Core Facility.

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Authors and Affiliations

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

    Kerstin A Sauer, Petra Scholtes, Roman Karwot & Susetta Finotto

  2. Asthma Core Facility, I Medical Clinic, University of Mainz, Germany

    Petra Scholtes & Susetta Finotto

Authors
  1. Kerstin A Sauer
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  2. Petra Scholtes
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  3. Roman Karwot
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  4. Susetta Finotto
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Correspondence to Susetta Finotto.

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Sauer, K., Scholtes, P., Karwot, R. et al. Isolation of CD4+ T cells from murine lungs: a method to analyze ongoing immune responses in the lung. Nat Protoc 1, 2870–2875 (2006). https://doi.org/10.1038/nprot.2006.435

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