Isolation and Identification of Conventional Dendritic Cell Subsets from the Intestine of Mice and Men

  • Charlotte L. ScottEmail author
  • Pamela B. WrightEmail author
  • Simon W. F. Milling
  • Allan McI Mowat
Part of the Methods in Molecular Biology book series (MIMB, volume 1423)


The identification of conventional dendritic cells (cDCs) in the intestinal mucosa has been hampered by the difficulties associated with isolating cells from the intestine and by the fact that overlapping markers have made it complicated to discriminate them accurately from other intestinal mononuclear phagocytes such as macrophages (MFs). Here we detail the protocols we have developed to isolate live leukocytes from both murine and human small and large intestines and describe reliable strategies which can be used to identify bona fide cDCs in such preparations.

Key words

Dendritic cells Subsets Isolation Identification Intestine 


  1. 1.
    Mowat AM (2003) Anatomical basis of tolerance and immunity to intestinal antigens. Nat Rev Immunol 3:331–341CrossRefPubMedGoogle Scholar
  2. 2.
    Scott CL, Murray TFPZ, Beckham KSH et al (2014) Signal Regulatory Protein alpha (SIRPα) regulates the homeostasis of CD103(+) CD11b(+) DCs in the intestinal lamina propria. Eur J Immunol. doi: 10.1002/eji.201444859 PubMedPubMedCentralGoogle Scholar
  3. 3.
    Persson EK, Uronen-Hansson H, Semmrich M et al (2013) IRF4 transcription-factor-dependent CD103(+)CD11b(+) dendritic cells drive mucosal T helper 17 cell differentiation. Immunity 38:958–969CrossRefPubMedGoogle Scholar
  4. 4.
    Schlitzer A, McGovern N, Teo P et al (2013) IRF4 transcription factor-dependent CD11b(+) dendritic cells in human and mouse control mucosal IL-17 cytokine responses. Immunity 38:970–983CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Cerovic V, Houston SA, Scott CL et al (2013) Intestinal CD103(-) dendritic cells migrate in lymph and prime effector T cells. Mucosal Immunol 6:104–113CrossRefPubMedGoogle Scholar
  6. 6.
    Scott CL, Bain CC, Wright PB et al (2014) CCR2(+)CD103(-) intestinal dendritic cells develop from DC-committed precursors and induce interleukin-17 production by T cells. Mucosal Immunol. doi: 10.1038/mi.2014.70 Google Scholar
  7. 7.
    Mowat AM, Agace WW (2014) Regional specialization within the intestinal immune system. Nat Rev Immunol 14:667–685CrossRefPubMedGoogle Scholar
  8. 8.
    Mayrhofer G, Pugh CW, Barclay AN (1983) The distribution, ontogeny and origin in the rat of Ia-positive cells with dendritic morphology and of Ia antigen in epithelia, with special reference to the intestine. Eur J Immunol 13:112–122CrossRefPubMedGoogle Scholar
  9. 9.
    Cerovic V, Bain CC, Mowat AM et al (2014) Intestinal macrophages and dendritic cells: what’s the difference? Trends Immunol 35:270–277CrossRefPubMedGoogle Scholar
  10. 10.
    Hadis U, Wahl B, Schulz O et al (2011) Intestinal tolerance requires gut homing and expansion of FoxP3+ regulatory T cells in the lamina propria. Immunity 34:237–246CrossRefPubMedGoogle Scholar
  11. 11.
    Guilliams M, Ginhoux F, Jakubzick C et al (2014) Dendritic cells, monocytes and macrophages: a unified nomenclature based on ontogeny. Nat Rev Immunol 14:571–578CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Schraml BU, van Blijswijk J, Zelenay S et al (2013) Genetic tracing via DNGR-1 expression history defines dendritic cells as a hematopoietic lineage. Cell 154:843–858CrossRefPubMedGoogle Scholar
  13. 13.
    Persson EK, Scott CL, Mowat AM et al (2013) Dendritic cell subsets in the intestinal lamina propria: ontogeny and function. Eur J Immunol 12:3098–3107CrossRefGoogle Scholar
  14. 14.
    Bachem A, Hartung E, Güttler S et al (2012) Expression of XCR1 characterizes the Batf3-dependent lineage of dendritic cells Capable of antigen cross-presentation. Front Immunol 3:214CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Hildner K, Edelson BT, Purtha WE et al (2008) Batf3 deficiency reveals a critical role for CD8alpha + dendritic cells in cytotoxic T cell immunity. Science (New York, NY) 322:1097–1100CrossRefGoogle Scholar
  16. 16.
    Cerovic V, Houston SA, Westlund J et al (2014) Lymph borne CD8a+ DCs are uniquely able to cross-prime CD8+ T cells with antigen acquired from intestinal epithelial cells. Mucosal Immunol. doi: 10.1038/mi.2014.40 Google Scholar
  17. 17.
    Lewis KL, Caton ML, Bogunovic M et al (2011) Notch2 receptor signaling controls functional differentiation of dendritic cells in the spleen and intestine. Immunity 35:780–791CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Satpathy AT, Briseño CG, Lee JS et al (2013) Notch2-dependent classical dendritic cells orchestrate intestinal immunity to attaching-and-effacing bacterial pathogens. Nat Immunol 14:937–948CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Welty NE, Staley C, Ghilardi N et al (2013) Intestinal lamina propria dendritic cells maintain T cell homeostasis but do not affect commensalism. J Exp Med 210:2011–2024CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Bachem A, Güttler S, Hartung E et al (2010) Superior antigen cross-presentation and XCR1 expression define human CD11c + CD141+ cells as homologues of mouse CD8+ dendritic cells. J Exp Med 207:1273–1281CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, College of Veterinary, Medical and Life SciencesUniversity of GlasgowScotlandUK
  2. 2.Laboratory of Mucosal Immunology and Immunoregulation, Inflammation Research Centre (IRC)VIB Ghent UniversityGhent (Zwijnaarde)Belgium
  3. 3.Department of Biomedical Molecular BiologyGhent University HospitalGhentBelgium

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