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High-resolution imaging of living gut mucosa: lymphocyte clusters beneath intestinal M cells are highly dynamic structures

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Abstract

In the Peyer’s patches of the small intestine, specialized epithelial cells, the membranous (M) cells, sample antigenic matter from the gut lumen and bring it into contact with cells of the immune system, which are then capable of initiating specific immune reactions. Using autofluorescence 2-photon (A2P) microscopy, we imaged living intestinal mucosa at a 0.5-μm resolution. We identified individual M cells without the aid of a marker and in vivo analyzed their sampling function over hours. Time-lapse recordings revealed that lymphocytes associated with M cells display a remarkable degree of motility with average speed rates of 8.2 μm/min, to form new M cell–associated lymphocyte clusters within less than 15 min. The lymphocytes drastically deform the M cells’ cytoplasm and laterally move from one lymphocyte cluster to the next. This implies that the micro-compartment beneath M cells is a highly efficient container to bring potentially harmful antigens into contact with large numbers of immunocompetent cells. Our setup opens a new window for high-resolution 3D imaging of functional processes occurring in lymphoid and mucosal tissues.

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Acknowledgments

We thank M. Hildner, H. Manfeldt and C. Örün for excellent technical assistance.

Funding

This study was supported by the German research foundation (DFG), Projects No.: Ge 647/9; Ge 647/10; HU 629/3; HU 629/4; INST 1757/15-1 FUGG.

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Author notes

  1. Tobias Fischer and Antje Klinger contributed equally to this work.

Authors and Affiliations

  1. Institute of Anatomy II, University Hospital Jena, Teichgraben 7, 07743, Jena, Germany

    Tobias Fischer, Falk Nitzsche, Torsten Bölke & Andreas Gebert

  2. Institute of Anatomy, University of Luebeck, Luebeck, Germany

    Antje Klinger, Dorthe von Smolinski & Andreas Gebert

  3. Institute of Biomedical Optics, University of Luebeck, Luebeck, Germany

    Regina Orzekowsky-Schroeder, Alfred Vogel & Gereon Hüttmann

Authors
  1. Tobias Fischer
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  2. Antje Klinger
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  3. Dorthe von Smolinski
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  4. Regina Orzekowsky-Schroeder
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  5. Falk Nitzsche
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  6. Torsten Bölke
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  7. Alfred Vogel
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  8. Gereon Hüttmann
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  9. Andreas Gebert
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Corresponding author

Correspondence to Tobias Fischer.

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The authors declare that there is no conflict of interest.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All animal experiments were approved by the Institutional Animal Experiment Committee of the university and received approval of the local authorities (Ministerium für Umwelt, Naturschutz und Landwirtschaft Schleswig-Holstein V742-72241.122 and Thüringer Landesamt für Verbraucherschutz 02/003/14).

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Fischer, T., Klinger, A., von Smolinski, D. et al. High-resolution imaging of living gut mucosa: lymphocyte clusters beneath intestinal M cells are highly dynamic structures. Cell Tissue Res 380, 539–546 (2020). https://doi.org/10.1007/s00441-020-03167-z

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  • DOI: https://doi.org/10.1007/s00441-020-03167-z

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