Skip to main content
SpringerLink
Log in

Isolation and Characterization of Innate Lymphoid Cells within the Murine Tumor Microenvironment

  • Protocol
  • First Online:
Innate Lymphoid Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2121))

Abstract

Innate lymphoid cells (ILCs) are important for both tissue immunity and tissue homeostasis. They are classified into three groups: Group 1 ILCs include NK cells, which are important in eliciting immunity against intracellular pathogens; group 2 ILCs protect against parasitic helminths; and group 3 ILCs protect against extracellular pathogens. The role of ILCs in cancer immunity remains unclear. In this chapter, we discuss methods for isolating and characterizing tumor-infiltrating ILC subsets within the tumor microenvironment in an experimental murine model of B16 melanoma. The chapter also highlights the expression of PD-1 on the various ILC subsets within the tumor microenvironment.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Salimi M, Barlow JL, Saunders SP et al (2013) A role for IL-25 and IL-33-driven type-2 innate lymphoid cells in atopic dermatitis. J Exp Med 210(13):2939–2950

    Article  CAS  Google Scholar 

  2. Geremia A, Arancibia-Carcamo CV, Fleming MP et al (2011) IL-23-responsive innate lymphoid cells are increased in inflammatory bowel disease. J Exp Med 208(6):1127–1133

    Article  CAS  Google Scholar 

  3. Spits H, Artis D, Colonna M et al (2013) Innate lymphoid cells—a proposal for uniform nomenclature. Nat Rev Immunol 13(2):145–149

    Article  CAS  Google Scholar 

  4. Crellin NK, Trifari S, Kaplan CD et al (2010) Regulation of cytokine secretion in human CD127(+) LTi-like innate lymphoid cells by Toll-like receptor 2. Immunity 33(5):752–764

    Article  CAS  Google Scholar 

  5. Herberman RB, Nunn ME, Holden HT et al (1975) Natural cytotoxic reactivity of mouse lymphoid cells against syngeneic and allogeneic tumors. II. Characterization of effector cells. Int J Cancer 16(2):230–239

    Article  CAS  Google Scholar 

  6. Weizman OE, Adams NM, Schuster IS et al (2017) ILC1 confer early host protection at initial sites of viral infection. Cell 171(4):795–808.e12

    Article  CAS  Google Scholar 

  7. Klose CSN, Kiss EA, Schwierzeck V et al (2014) Differentiation of type 1 ILCs from a common progenitor to all helper-like innate lymphoid cell lineages. Cell 157(2):340–356

    Article  CAS  Google Scholar 

  8. Neill DR, Wong SH, Bellosi A et al (2010) Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity. Nature 464(7293):1367–1370

    Article  CAS  Google Scholar 

  9. Huang Y, Guo L, Qiu J et al (2015) IL-25-responsive, lineage-negative KLRG1(hi) cells are multipotential ‘inflammatory’ type 2 innate lymphoid cells. Nat Immunol 16(2):161–169

    Article  CAS  Google Scholar 

  10. Sanos SL, Bui VL, Mortha A et al (2009) RORgammat and commensal microflora are required for the differentiation of mucosal interleukin 22-producing NKp46+ cells. Nat Immunol 10(1):83–91

    Article  CAS  Google Scholar 

  11. Cella M, Fuchs A, Vermi W et al (2009) A human natural killer cell subset provides an innate source of IL-22 for mucosal immunity. Nature 457(7230):722–725

    Article  CAS  Google Scholar 

  12. Buonocore S, Ahern PP, Uhlig HH et al (2010) Innate lymphoid cells drive interleukin-23-dependent innate intestinal pathology. Nature 464(7293):1371–1375

    Article  CAS  Google Scholar 

  13. Sonnenberg GF, Monticelli LA, Ellosi MM et al (2011) CD4(+) lymphoid tissue-inducer cells promote innate immunity in the gut. Immunity 34(1):122–134

    Article  CAS  Google Scholar 

  14. Takatori H, Kanno Y, Watford WT et al (2009) Lymphoid tissue inducer-like cells are an innate source of IL-17 and IL-22. J Exp Med 206(1):35–41

    Article  CAS  Google Scholar 

  15. Morvan MG, Lanier LL (2016) NK cells and cancer: you can teach innate cells new tricks. Nat Rev Cancer 16(1):7–19

    Article  CAS  Google Scholar 

  16. Martini M, Testi MG, Passetto M et al (2010) IFN-gamma-mediated upmodulation of MHC class I expression activates tumor-specific immune response in a mouse model of prostate cancer. Vaccine 28(20):3548–3557

    Article  CAS  Google Scholar 

  17. Nakajima C, Uekusa Y, Iwasaki M et al (2001) A role of interferon-gamma (IFN-gamma) in tumor immunity: T cells with the capacity to reject tumor cells are generated but fail to migrate to tumor sites in IFN-gamma-deficient mice. Cancer Res 61(8):3399–3405

    CAS  PubMed  Google Scholar 

  18. Beatty G, Paterson Y (2001) IFN-gamma-dependent inhibition of tumor angiogenesis by tumor-infiltrating CD4+ T cells requires tumor responsiveness to IFN-gamma. J Immunol 166(4):2276–2282

    Article  CAS  Google Scholar 

  19. Larmonier N, Cathelin D, Larmonier C et al (2007) The inhibition of TNF-alpha anti-tumoral properties by blocking antibodies promotes tumor growth in a rat model. Exp Cell Res 313(11):2345–2355

    Article  CAS  Google Scholar 

  20. Jovanovic IP, Pejnovic N, Radosavljevic GD et al (2014) Interleukin-33/ST2 axis promotes breast cancer growth and metastases by facilitating intratumoral accumulation of immunosuppressive and innate lymphoid cells. Int J Cancer 134(7):1669–1682

    Article  CAS  Google Scholar 

  21. Molofsky AB, Nussbaum JC, Liang HE et al (2013) Innate lymphoid type 2 cells sustain visceral adipose tissue eosinophils and alternatively activated macrophages. J Exp Med 210(3):535–549

    Article  CAS  Google Scholar 

  22. Huber S, Gagliani N, Zenewicz LA et al (2012) IL-22BP is regulated by the inflammasome and modulates tumorigenesis in the intestine. Nature 491(7423):259–263

    Article  CAS  Google Scholar 

  23. Chan IH, Jain R, Tessmer MS et al (2014) Interleukin-23 is sufficient to induce rapid de novo gut tumorigenesis, independent of carcinogens, through activation of innate lymphoid cells. Mucosal Immunol 7(4):842–856

    Article  CAS  Google Scholar 

  24. Schatton T, Murphy GF, Frank NY et al (2008) Identification of cells initiating human melanomas. Nature 451(7176):345–349

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank the Academy of Medical Sciences, Springboard Award [SBF003\1129] supported by Wellcome Trust and Newcastle University Research Fellowship for supporting this work. G.M. is supported by a Medical Research Council [MRC] Discovery Medicine North Ph.D. studentship. M.P. is supported by a Wellcome Trust Vacation Scholarship [216503/Z/19/Z].

Author information

Authors and Affiliations

  1. Laboratory of T cell Regulation, Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, UK

    Grace Mallett, Warren Patterson, Megan Payne & Shoba Amarnath

Authors
  1. Grace Mallett
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Warren Patterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Megan Payne
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shoba Amarnath
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shoba Amarnath .

Editor information

Editors and Affiliations

  1. Laboratory of T cell Regulation, Institute of Cellular Medicine, Newcastle University, Newcastle Upon Tyne, UK

    Shoba Amarnath

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Mallett, G., Patterson, W., Payne, M., Amarnath, S. (2020). Isolation and Characterization of Innate Lymphoid Cells within the Murine Tumor Microenvironment. In: Amarnath, S. (eds) Innate Lymphoid Cells . Methods in Molecular Biology, vol 2121. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0338-3_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0338-3_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0337-6

  • Online ISBN: 978-1-0716-0338-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation