Guidelines for mouse and human DC functional assays

European Journal of Immunology, vol. 53, pp. e2249925

Abstract

This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy, and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs and various non-lymphoid tissues. Recent studies have provided evidence for an increasing number of phenotypically distinct conventional DC (cDC) subsets that on one hand exhibit a certain functional plasticity, but on the other hand are characterized by their tissue- and context-dependent functional specialization. Here, we describe a selection of assays for the functional characterization of mouse and human cDC. The first two protocols illustrate analysis of cDC endocytosis and metabolism, followed by guidelines for transcriptomic and proteomic characterization of cDC populations. Then, a larger group of assays describes the characterization of cDC migration in vitro, ex vivo, and in vivo. The final guidelines measure cDC inflammasome and antigen (cross)-presentation activity. While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all co-authors, making it an essential resource for basic and clinical DC immunologists.

Authors 46

1. Björn E. Clausen corresponding

   Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

   Affiliation as printed

   Institute for Molecular Medicine Paul Klein Center for Immune Intervention University Medical Center of the Johannes Gutenberg‐University Mainz Mainz Germany

   Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

   Institute for Molecular Medicine, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany

   Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

   Section lead authors

2. Lukas Amon

   Universitätsklinikum Erlangen

   Affiliation as printed

   Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

   Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

3. Ronald A. Backer

   Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

   Affiliation as printed

   Institute for Molecular Medicine Paul Klein Center for Immune Intervention University Medical Center of the Johannes Gutenberg‐University Mainz Mainz Germany

   Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

   Institute for Molecular Medicine, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany

   Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

4. Luciana Berod

   Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

   Affiliation as printed

   Institute of Molecular Medicine University Medical Center of the Johannes Gutenberg‐University Mainz Germany

   Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

   Institute of Molecular Medicine, University Medical Center of the Johannes Gutenberg-University Mainz, Germany

   Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

   Section lead authors

5. Tobias Bopp

   Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

   Affiliation as printed

   Institute of Immunology Paul Klein Center for Immune Intervention University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

   Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

   Institute of Immunology, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

   Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

6. Anna Brand

   Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

   Affiliation as printed

   Institute for Molecular Medicine Paul Klein Center for Immune Intervention University Medical Center of the Johannes Gutenberg‐University Mainz Mainz Germany

   Institute for Molecular Medicine, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany

7. Sven Burgdorf

   University of Bonn

   Affiliation as printed

   Laboratory of Cellular Immunology LIMES Institute University of Bonn Bonn Germany

   Laboratory of Cellular Immunology, LIMES Institute, University of Bonn, Bonn, Germany

   Section lead authors

8. Luxia Chen

   University Hospital Heidelberg

   Affiliation as printed

   Department of Dermatology University Hospital Heidelberg Heidelberg Germany

   Department of Dermatology, University Hospital Heidelberg, Heidelberg, Germany

9. Meihong Da

   University Hospital Heidelberg

   Affiliation as printed

   Department of Dermatology University Hospital Heidelberg Heidelberg Germany

   Department of Dermatology, University Hospital Heidelberg, Heidelberg, Germany

10. Ute Distler

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Immunology Paul Klein Center for Immune Intervention University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Institute of Immunology, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Section lead authors

11. Regine J. Dress

    University Medical Center Hamburg-Eppendorf

    Affiliation as printed

    Institute of Systems Immunology Hamburg Center for Translational Immunology (HCTI) University Medical Center Hamburg‐Eppendorf Hamburg Germany

    Institute of Systems Immunology, Hamburg Center for Translational Immunology (HCTI), University Medical Center Hamburg-Eppendorf, Hamburg, Germany

12. Diana Dudziak

    Friedrich-Alexander-Universität Erlangen-Nürnberg · Universitätsklinikum Erlangen · Immunologie-Zentrum Zürich

    Affiliation as printed

    Deutsches Zentrum Immuntherapie (DZI) Germany

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Medical Immunology Campus Erlangen (MICE) Erlangen Germany

    Deutsches Zentrum Immuntherapie (DZI), Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

    Medical Immunology Campus Erlangen (MICE), Erlangen, Germany

    Section lead authors

13. Charles‐Antoine Dutertre

    Inserm · Institut Gustave Roussy · La Ligue Contre le Cancer

    Affiliation as printed

    Gustave Roussy Cancer Campus Villejuif France

    Institut National de la Santé et de la Recherche Médicale (INSERM) U1015 Equipe Labellisée—Ligue Nationale contre le Cancer Villejuif France

    Gustave Roussy Cancer Campus, Villejuif, France

14. Christina Eich

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute for Molecular Medicine Paul Klein Center for Immune Intervention University Medical Center of the Johannes Gutenberg‐University Mainz Mainz Germany

    Institute for Molecular Medicine, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany

15. Anna Gabele

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Immunology Paul Klein Center for Immune Intervention University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Institute of Immunology, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

16. Melanie Geiger Aachen Institute for Biomedical Engineering Department of Cell Biology Institute for Biomedical Engineering Department of Cell Biology

    RWTH Aachen University

    Affiliation as printed

    Helmholtz‐Institute for Biomedical Engineering RWTH Aachen University Aachen Germany

    Institute for Biomedical Engineering Department of Cell Biology RWTH Aachen University Medical Faculty Aachen Germany

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany

    Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University, Medical Faculty, Aachen, Germany

17. Florent Ginhoux

    Agency for Science, Technology and Research · Inserm · Shanghai Jiao Tong University · Singapore Immunology Network · Institut Gustave Roussy · La Ligue Contre le Cancer · SingHealth Duke-NUS Academic Medical Centre · Shanghai Institute of Immunology

    Affiliation as printed

    Gustave Roussy Cancer Campus Villejuif France

    Institut National de la Santé et de la Recherche Médicale (INSERM) U1015 Equipe Labellisée—Ligue Nationale contre le Cancer Villejuif France

    Shanghai Institute of Immunology Department of Immunology and Microbiology Shanghai Jiao Tong University School of Medicine Shanghai China

    Singapore Immunology Network (SIgN), Agency for Science Technology and Research Singapore Singapore

    Translational Immunology Institute SingHealth Duke‐NUS Academic Medical Centre Singapore Singapore

    Gustave Roussy Cancer Campus, Villejuif, France

    Section lead authors

    Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Singapore Immunology Network (SIgN), Agency for Science, Technology and Research, Singapore, Singapore

    Translational Immunology Institute, SingHealth Duke-NUS Academic Medical Centre, Singapore, Singapore

18. Lucila Giusiano

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Medical Microbiology and Hygiene University Medical Center of the Johannes Gutenberg‐University Mainz Germany

    Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Germany

19. Gloria J. Godoy

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Medical Microbiology and Hygiene University Medical Center of the Johannes Gutenberg‐University Mainz Germany

    Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Germany

20. Ahmed E.I. Hamouda Aachen Institute for Biomedical Engineering Department of Cell Biology Institute for Biomedical Engineering Department of Cell Biology

    RWTH Aachen University

    Affiliation as printed

    Helmholtz‐Institute for Biomedical Engineering RWTH Aachen University Aachen Germany

    Institute for Biomedical Engineering Department of Cell Biology RWTH Aachen University Medical Faculty Aachen Germany

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany

    Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University, Medical Faculty, Aachen, Germany

21. Lukas Hatscher

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

22. Lukas Heger

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

23. Gordon F. Heidkamp

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

24. Lola C. Hernandez

    University Medical Center Hamburg-Eppendorf

    Affiliation as printed

    Cell Communication and Migration Laboratory Institute of Biochemistry and Molecular Cell Biology Center for Experimental Medicine University Medical Center Hamburg‐Eppendorf Hamburg Germany

    Cell Communication and Migration Laboratory, Institute of Biochemistry and Molecular Cell Biology, Center for Experimental Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

25. Lukas Jacobi

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

26. Tomasz Kaszubowski

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

27. Wan Ting Kong

    Inserm · Institut Gustave Roussy · La Ligue Contre le Cancer

    Affiliation as printed

    Gustave Roussy Cancer Campus Villejuif France

    Institut National de la Santé et de la Recherche Médicale (INSERM) U1015 Equipe Labellisée—Ligue Nationale contre le Cancer Villejuif France

    Gustave Roussy Cancer Campus, Villejuif, France

28. Christian H.K. Lehmann

    Friedrich-Alexander-Universität Erlangen-Nürnberg · Universitätsklinikum Erlangen · Immunologie-Zentrum Zürich

    Affiliation as printed

    Deutsches Zentrum Immuntherapie (DZI) Germany

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Medical Immunology Campus Erlangen (MICE) Erlangen Germany

    Deutsches Zentrum Immuntherapie (DZI), Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

    Medical Immunology Campus Erlangen (MICE), Erlangen, Germany

29. Tamara López‐López

    University Medical Center Hamburg-Eppendorf

    Affiliation as printed

    Cell Communication and Migration Laboratory Institute of Biochemistry and Molecular Cell Biology Center for Experimental Medicine University Medical Center Hamburg‐Eppendorf Hamburg Germany

    Cell Communication and Migration Laboratory, Institute of Biochemistry and Molecular Cell Biology, Center for Experimental Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

30. Karsten Mahnke

    University Hospital Heidelberg

    Affiliation as printed

    Department of Dermatology University Hospital Heidelberg Heidelberg Germany

    Department of Dermatology, University Hospital Heidelberg, Heidelberg, Germany

    Section lead authors

31. Dominik Nitsche

    University of Bonn

    Affiliation as printed

    Laboratory of Cellular Immunology LIMES Institute University of Bonn Bonn Germany

    Laboratory of Cellular Immunology, LIMES Institute, University of Bonn, Bonn, Germany

32. Jörg Renkawitz

    LMU Klinikum · Institut für Kardiovaskuläre Physiologie und Pathophysiologie · Ludwig-Maximilians-Universität München

    Affiliation as printed

    Biomedical Center (BMC) Walter Brendel Center of Experimental Medicine Institute of Cardiovascular Physiology and Pathophysiology Klinikum der Universität LMU Munich Munich Germany

    Section lead authors

33. Rifat A. Reza

    LMU Klinikum · Institut für Kardiovaskuläre Physiologie und Pathophysiologie · Ludwig-Maximilians-Universität München

    Affiliation as printed

    Biomedical Center (BMC) Walter Brendel Center of Experimental Medicine Institute of Cardiovascular Physiology and Pathophysiology Klinikum der Universität LMU Munich Munich Germany

34. Pablo José Sáez

    University Medical Center Hamburg-Eppendorf

    Affiliation as printed

    Cell Communication and Migration Laboratory Institute of Biochemistry and Molecular Cell Biology Center for Experimental Medicine University Medical Center Hamburg‐Eppendorf Hamburg Germany

    Cell Communication and Migration Laboratory, Institute of Biochemistry and Molecular Cell Biology, Center for Experimental Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Section lead authors

35. Laura Schlautmann

    University of Bonn

    Affiliation as printed

    Laboratory of Cellular Immunology LIMES Institute University of Bonn Bonn Germany

    Laboratory of Cellular Immunology, LIMES Institute, University of Bonn, Bonn, Germany

36. Madeleine T. Schmitt

    LMU Klinikum · Institut für Kardiovaskuläre Physiologie und Pathophysiologie · Ludwig-Maximilians-Universität München

    Affiliation as printed

    Biomedical Center (BMC) Walter Brendel Center of Experimental Medicine Institute of Cardiovascular Physiology and Pathophysiology Klinikum der Universität LMU Munich Munich Germany

37. Anna Seichter

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

38. Malte Sielaff

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Immunology Paul Klein Center for Immune Intervention University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Institute of Immunology, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

39. Tim D. Sparwasser

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz

    Affiliation as printed

    Institute of Medical Microbiology and Hygiene University Medical Center of the Johannes Gutenberg‐University Mainz Germany

    Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Institute of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg-University Mainz, Germany

    Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

40. Patrizia Stoitzner

    Innsbruck Medical University · Universität Innsbruck

    Affiliation as printed

    Department of Dermatology Venerology & Allergology Medical University Innsbruck Innsbruck Austria

    Department of Dermatology, Venerology & Allergology, Medical University Innsbruck, Innsbruck, Austria

41. Giorgi Tchitashvili

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

42. Stefan Tenzer

    Johannes Gutenberg University Mainz · University Medical Center of the Johannes Gutenberg University Mainz · Helmholtz Institute for Translational Oncology Mainz

    Affiliation as printed

    Helmholtz Institute for Translational Oncology Mainz (HI‐TRON Mainz) Mainz Germany

    Institute of Immunology Paul Klein Center for Immune Intervention University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Research Center for Immunotherapy (FZI) University Medical Center of the Johannes‐Gutenberg University Mainz Mainz Germany

    Helmholtz Institute for Translational Oncology Mainz (HI-TRON Mainz), Mainz, Germany

    Institute of Immunology, Paul Klein Center for Immune Intervention, University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Research Center for Immunotherapy (FZI), University Medical Center of the Johannes-Gutenberg University Mainz, Mainz, Germany

    Section lead authors

43. Nounagnon Romaric Tochoedo

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

44. Damir Vurnek

    Universitätsklinikum Erlangen

    Affiliation as printed

    Laboratory of Dendritic Cell Biology Department of Dermatology University Hospital Erlangen Germany

    Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Germany

45. Fabian Zink

    University of Bonn

    Affiliation as printed

    Laboratory of Cellular Immunology LIMES Institute University of Bonn Bonn Germany

    Laboratory of Cellular Immunology, LIMES Institute, University of Bonn, Bonn, Germany

46. Thomas Hieronymus corresponding Aachen Institute for Biomedical Engineering Department of Cell Biology Institute of Cell and Tumor Biology Institute for Biomedical Engineering Department of Cell Biology

    RWTH Aachen University

    Affiliation as printed

    Helmholtz‐Institute for Biomedical Engineering RWTH Aachen University Aachen Germany

    Institute for Biomedical Engineering Department of Cell Biology RWTH Aachen University Medical Faculty Aachen Germany

    Institute of Cell and Tumor Biology RWTH Aachen University Medical Faculty Germany

    Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany

    Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University, Medical Faculty, Aachen, Germany

    Institute of Cell and Tumor Biology, RWTH Aachen University, Medical Faculty, Germany

    Section lead authors