Skip to main content
SpringerLink
Log in

CD107a Degranulation Assay to Evaluate Immune Cell Antitumor Activity

  • Protocol
  • First Online:
Cancer Immunosurveillance

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1884))

Abstract

Cancer development is under surveillance by the immune system of the host. Tumor cells can be recognized and killed by cytotoxic lymphocytes— such as CD8+ T lymphocytes and natural killer (NK) cells—mainly through the immune secretion of lytic granules that kill target cells. This process involves the fusion of the granule membrane with the cytoplasmic membrane of the immune effector cell, resulting in surface exposure of lysosomal-associated proteins that are typically present on the lipid bilayer surrounding lytic granules, such as CD107a. Therefore, membrane expression of CD107a constitutes a marker of immune cell activation and cytotoxic degranulation. In this chapter, we detail the steps required to isolate peripheral blood mononuclear cells (PBMCs), coculture them with target tumor cell lines, and evaluate the cytotoxic immune function by means of flow cytometry evaluation of CD107a expression on the surface of NK cells.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
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. Lopez-Soto A, Gonzalez S, Smyth MJ, Galluzzi L (2017) Control of metastasis by NK cells. Cancer Cell 32(2):135–154. https://doi.org/10.1016/j.ccell.2017.06.009

    Article  CAS  PubMed  Google Scholar 

  2. Gonzalez S, Lopez-Soto A, Suarez-Alvarez B, Lopez-Vazquez A, Lopez-Larrea C (2008) NKG2D ligands: key targets of the immune response. Trends Immunol 29(8):397–403. https://doi.org/10.1016/j.it.2008.04.007

    Article  CAS  PubMed  Google Scholar 

  3. Nausch N, Cerwenka A (2008) NKG2D ligands in tumor immunity. Oncogene 27(45):5944–5958. https://doi.org/10.1038/onc.2008.272

    Article  CAS  PubMed  Google Scholar 

  4. Lopez-Soto A, Huergo-Zapico L, Acebes-Huerta A, Villa-Alvarez M, Gonzalez S (2015) NKG2D signaling in cancer immunosurveillance. Int J Cancer 136(8):1741–1750. https://doi.org/10.1002/ijc.28775

    Article  CAS  PubMed  Google Scholar 

  5. Lieberman J (2003) The ABCs of granule-mediated cytotoxicity: new weapons in the arsenal. Nat Rev Immunol 3(5):361–370. https://doi.org/10.1038/nri1083

    Article  CAS  PubMed  Google Scholar 

  6. Uhrberg M (2005) The CD107 mobilization assay: viable isolation and immunotherapeutic potential of tumor-cytolytic NK cells. Leukemia 19(5):707–709. https://doi.org/10.1038/sj.leu.2403705

    Article  CAS  PubMed  Google Scholar 

  7. Terasawa K, Tomabechi Y, Ikeda M, Ehara H, Kukimoto-Niino M, Wakiyama M, Podyma-Inoue KA, Rajapakshe AR, Watabe T, Shirouzu M, Hara-Yokoyama M (2016) Lysosome-associated membrane proteins-1 and -2 (LAMP-1 and LAMP-2) assemble via distinct modes. Biochem Biophys Res Commun 479(3):489–495. https://doi.org/10.1016/j.bbrc.2016.09.093

    Article  CAS  PubMed  Google Scholar 

  8. Cohnen A, Chiang SC, Stojanovic A, Schmidt H, Claus M, Saftig P, Janssen O, Cerwenka A, Bryceson YT, Watzl C (2013) Surface CD107a/LAMP-1 protects natural killer cells from degranulation-associated damage. Blood 122(8):1411–1418. https://doi.org/10.1182/blood-2012-07-441832

    Article  CAS  PubMed  Google Scholar 

  9. Betts MR, Brenchley JM, Price DA, De Rosa SC, Douek DC, Roederer M, Koup RA (2003) Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J Immunol Methods 281(1–2):65–78. https://doi.org/10.1016/S0022-1759(03)00265-5

    Article  CAS  PubMed  Google Scholar 

  10. Alter G, Malenfant JM, Altfeld M (2004) CD107a as a functional marker for the identification of natural killer cell activity. J Immunol Methods 294(1–2):15–22. https://doi.org/10.1016/j.jim.2004.08.008

    Article  CAS  PubMed  Google Scholar 

  11. Penack O, Gentilini C, Fischer L, Asemissen AM, Scheibenbogen C, Thiel E, Uharek L (2005) CD56dimCD16neg cells are responsible for natural cytotoxicity against tumor targets. Leukemia 19(5):835–840. https://doi.org/10.1038/sj.leu.2403704

    Article  CAS  PubMed  Google Scholar 

  12. Aktas E, Kucuksezer UC, Bilgic S, Erten G, Deniz G (2009) Relationship between CD107a expression and cytotoxic activity. Cell Immunol 254(2):149–154. https://doi.org/10.1016/j.cellimm.2008.08.007

    Article  CAS  PubMed  Google Scholar 

  13. Kannan K, Stewart RM, Bounds W, Carlsson SR, Fukuda M, Betzing KW, Holcombe RF (1996) Lysosome-associated membrane proteins h-LAMP1 (CD107a) and h-LAMP2 (CD107b) are activation-dependent cell surface glycoproteins in human peripheral blood mononuclear cells which mediate cell adhesion to vascular endothelium. Cell Immunol 171(1):10–19. https://doi.org/10.1006/cimm.1996.0167

    Article  CAS  PubMed  Google Scholar 

  14. Lopez-Soto A, Huergo-Zapico L, Galvan JA, Rodrigo L, de Herreros AG, Astudillo A, Gonzalez S (2013) Epithelial-mesenchymal transition induces an antitumor immune response mediated by NKG2D receptor. J Immunol 190(8):4408–4419. https://doi.org/10.4049/jimmunol.1202950

    Article  CAS  PubMed  Google Scholar 

  15. Acebes-Huerta A, Lorenzo-Herrero S, Folgueras AR, Huergo-Zapico L, Lopez-Larrea C, Lopez-Soto A, Gonzalez S (2016) Drug-induced hyperploidy stimulates an antitumor NK cell response mediated by NKG2D and DNAM-1 receptors. Oncoimmunology 5(2):e1074378. https://doi.org/10.1080/2162402X.2015.1074378

    Article  CAS  PubMed  Google Scholar 

  16. Bryceson YT, Pende D, Maul-Pavicic A, Gilmour KC, Ufheil H, Vraetz T, Chiang SC, Marcenaro S, Meazza R, Bondzio I, Walshe D, Janka G, Lehmberg K, Beutel K, zur Stadt U, Binder N, Arico M, Moretta L, Henter JI, Ehl S (2012) A prospective evaluation of degranulation assays in the rapid diagnosis of familial hemophagocytic syndromes. Blood 119(12):2754–2763. https://doi.org/10.1182/blood-2011-08-374199

    Article  CAS  PubMed  Google Scholar 

  17. Ehl S, de Saint BG (2014) Chapter 20 - Genetic diseases predisposing to HLH. In: Stiehm's immune deficiencies. Academic Press, Amsterdam, pp 437–460. https://doi.org/10.1016/B978-0-12-405546-9.00020-0

    Chapter  Google Scholar 

  18. Rubin TS, Zhang K, Gifford C, Lane A, Choo S, Bleesing JJ, Marsh RA (2017) Perforin and CD107a testing is superior to NK cell function testing for screening patients for genetic HLH. Blood 129(22):2993–2999. https://doi.org/10.1182/blood-2016-12-753830

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by a Spanish grant of Instituto de Salud Carlos III (PI16/01485). S.L.H. holds a predoctoral Severo Ochoa Fellowship (BP14–150) from FICYT of Principado de Asturias, Spain.

Author information

Authors and Affiliations

  1. Departamento de Biología Funcional, Inmunología, Universidad of Oviedo, Oviedo, Spain

    Seila Lorenzo-Herrero, Christian Sordo-Bahamonde & Segundo Gonzalez

  2. Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Instituto de Investigación Sanitaria del Principado de Asturias (IISPA), Oviedo, Spain

    Seila Lorenzo-Herrero, Christian Sordo-Bahamonde & Segundo Gonzalez

  3. Departamento de Biología Funcional, Inmunología, Universidad de Oviedo, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Oviedo, Spain

    Alejandro López-Soto

Authors
  1. Seila Lorenzo-Herrero
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christian Sordo-Bahamonde
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Segundo Gonzalez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alejandro López-Soto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alejandro López-Soto .

Editor information

Editors and Affiliations

  1. Departamento de Biología Funcional, Inmunología, Universidad de Oviedo, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Oviedo, Spain

    Alejandro López-Soto

  2. Departamento de Bioquímica y Biología Molecular, Universidad de Oviedo, Instituto Universitario de Oncología del Principado de Asturias (IUOPA), Oviedo, Asturias, Spain

    Alicia R. Folgueras

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lorenzo-Herrero, S., Sordo-Bahamonde, C., Gonzalez, S., López-Soto, A. (2019). CD107a Degranulation Assay to Evaluate Immune Cell Antitumor Activity. In: López-Soto, A., Folgueras, A. (eds) Cancer Immunosurveillance. Methods in Molecular Biology, vol 1884. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8885-3_7

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8885-3_7

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8884-6

  • Online ISBN: 978-1-4939-8885-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation