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Isolation and Flow Cytometry Analysis of Innate Lymphoid Cells from the Intestinal Lamina Propria

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Inflammation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1559))

Abstract

The intestinal mucosa constitutes the biggest surface area of the body. It is constantly challenged by bacteria, commensal and pathogenic, protozoa, and food-derived irritants. In order to maintain homeostasis, a complex network of signaling circuits has evolved that includes contributions of immune cells. In recent years a subset of lymphocytes, which belong to the innate immune system, has caught particular attention. These so-called innate lymphoid cells (ILC) reside within the lamina propria of the small and large intestines and rapidly respond to environmental challenges. They provide immunity to various types of infections but may also contribute to organ homeostasis as they produce factors acting on epithelial cells thereby enhancing barrier integrity. Here, we describe how these cells can be isolated from their environment and provide an in-depth protocol how to visualize the various ILC subsets by flow cytometry.

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References

  1. Belkaid Y, Hand TW (2014) Role of the microbiota in immunity and inflammation. Cell 157:121–141. doi:10.1016/j.cell.2014.03.011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Mowat AM, Agace WW (2014) Regional specialization within the intestinal immune system. Nat Rev Immunol 14:667–685. doi:10.1038/nri3738

    Article  CAS  PubMed  Google Scholar 

  3. Walker JA, Barlow JL, McKenzie ANJ (2013) Innate lymphoid cells—how did we miss them? Nat Rev Immunol 13:75–87. doi:10.1038/nri3349

    Article  CAS  PubMed  Google Scholar 

  4. Mebius RE et al (2001) The fetal liver counterpart of adult common lymphoid progenitors gives rise to all lymphoid lineages, CD45 + CD4 + CD3− cells, as well as macrophages. J Immunol 166:6593–6601

    Article  CAS  PubMed  Google Scholar 

  5. Klose CSN et al (2014) Differentiation of type 1 ILCs from a common progenitor to all helper-like innate lymphoid cell lineages. Cell 157:340–356. doi:10.1016/j.cell.2014.03.030

    Article  CAS  PubMed  Google Scholar 

  6. Diefenbach A, Colonna M, Koyasu S (2014) Development, differentiation, and diversity of innate lymphoid cells. Immunity 41:354–365. doi:10.1016/j.immuni.2014.09.005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Wong SH et al (2012) Transcription factor RORα is critical for nuocyte development. Nat Immunol 13:229–236. doi:10.1038/ni.2208

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Hoyler T et al (2012) The transcription factor GATA-3 controls cell fate and maintenance of type 2 innate lymphoid cells. Immunity 37:634–648. doi:10.1016/j.immuni.2012.06.020

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Moro K et al (2009) Innate production of TH2 cytokines by adipose tissue-associated c-Kit + Sca-1+ lymphoid cells. Nature 463:540–544. doi:10.1038/nature08636

    Article  PubMed  Google Scholar 

  10. Neill DR et al (2010) Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464:1367–1370. doi:10.1038/nature08900

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Cella M et al (2009) A human natural killer cell subset provides an innate source of IL-22 for mucosal immunity. Nature 457:722–725. doi:10.1038/nature07537

    Article  CAS  PubMed  Google Scholar 

  12. Sonnenberg GF, Monticelli LA, Elloso MM, Fouser LA, Artis D (2011) CD4(+) lymphoid tissue-inducer cells promote innate immunity in the gut. Immunity 34:122–134. doi:10.1016/j.immuni.2010.12.009

    Article  CAS  PubMed  Google Scholar 

  13. Sonnenberg GF, Fouser LA, Artis D (2011) Border patrol: regulation of immunity, inflammation and tissue homeostasis at barrier surfaces by IL-22. Nat Immunol 12:383–390. doi:10.1038/ni.2025

    Article  CAS  PubMed  Google Scholar 

  14. Goto Y et al (2014) Innate lymphoid cells regulate intestinal epithelial cell glycosylation. Science (New York, NY) 345:1254009. doi:10.1126/science.1254009

    Article  Google Scholar 

  15. Gordon SM et al (2012) The transcription factors T-bet and Eomes control key checkpoints of natural killer cell maturation. Immunity 36:55–67. doi:10.1016/j.immuni.2011.11.016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Sun JC, Lanier LL (2011) NK cell development, homeostasis and function: parallels with CD8+ T cells. Nat Rev Immunol 11:645–657. doi:10.1038/nri3044

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Vonarbourg C et al (2010) Regulated expression of nuclear receptor RORγt confers distinct functional fates to NK cell receptor-expressing RORγt(+) innate lymphocytes. Immunity 33:736–751. doi:10.1016/j.immuni.2010.10.017

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Andreas Diefenbach .

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Gronke, K., Kofoed-Nielsen, M., Diefenbach, A. (2017). Isolation and Flow Cytometry Analysis of Innate Lymphoid Cells from the Intestinal Lamina Propria. In: Clausen, B., Laman, J. (eds) Inflammation. Methods in Molecular Biology, vol 1559. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6786-5_17

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  • DOI: https://doi.org/10.1007/978-1-4939-6786-5_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6784-1

  • Online ISBN: 978-1-4939-6786-5

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