Skip to main content

Measuring Suppressive Activity and Autophagy in Myeloid-Derived Suppressor Cells

  • Protocol
  • First Online:
Myeloid-Derived Suppressor Cells

Abstract

Myeloid-derived suppressor cells (MDSC) are potent suppressor cells that accumulate in tumor microenvironment and inhibit anti-tumor responses. Assessment of cell-autonomous MDSC responses allows the precise characterization of MDSCs in various disease settings and elucidates the underlying mechanisms of MDSC-mediated immune suppression. Here we describe a protocol for the isolation of tumor infiltrating or splenic MDSC, as well as their subpopulations, from melanoma-inoculated mice using Fluorescent Activated Cell Sorting (FACS). We further provide protocols for investigation of the autophagy pathway and ex vivo assessment of MDSC suppressive function using lymph node responder cells. These assays allow a comprehensive characterization of MDSC in murine experimental models.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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

Similar content being viewed by others

References

  1. Hatziioannou A, Alissafi T, Verginis P (2017) Myeloid-derived suppressor cells and T regulatory cells in tumors: unraveling the dark side of the force. J Leukoc Biol 102(2):407–421. https://doi.org/10.1189/jlb.5VMR1116-493R

    Article  CAS  PubMed  Google Scholar 

  2. Gabrilovich DI, Ostrand-Rosenberg S, Bronte V (2012) Coordinated regulation of myeloid cells by tumours. Nat Rev Immunol 12(4):253–268. https://doi.org/10.1038/nri3175

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Veglia F, Perego M, Gabrilovich D (2018) Myeloid-derived suppressor cells coming of age. Nat Immunol 19(2):108–119. https://doi.org/10.1038/s41590-017-0022-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Weber R, Fleming V, Hu X, Nagibin V, Groth C, Altevogt P, Utikal J, Umansky V (2018) Myeloid-derived suppressor cells hinder the anti-cancer activity of immune checkpoint inhibitors. Front Immunol 9:1310. https://doi.org/10.3389/fimmu.2018.01310

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Ioannou M, Alissafi T, Lazaridis I, Deraos G, Matsoukas J, Gravanis A, Mastorodemos V, Plaitakis A, Sharpe A, Boumpas D, Verginis P (2012) Crucial role of granulocytic myeloid-derived suppressor cells in the regulation of central nervous system autoimmune disease. J Immunol 188(3):1136–1146. https://doi.org/10.4049/jimmunol.1101816

    Article  CAS  PubMed  Google Scholar 

  6. Ioannou M, Alissafi T, Boon L, Boumpas D, Verginis P (2013) In vivo ablation of plasmacytoid dendritic cells inhibits autoimmunity through expansion of myeloid-derived suppressor cells. J Immunol 190(6):2631–2640. https://doi.org/10.4049/jimmunol.1201897

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Vlachou K, Mintzas K, Glymenaki M, Ioannou M, Papadaki G, Bertsias GK, Sidiropoulos P, Boumpas DT, Verginis P (2016) Elimination of granulocytic myeloid-derived suppressor cells in lupus-prone mice linked to reactive oxygen species-dependent extracellular trap formation. Arthritis Rheumatol 68(2):449–461. https://doi.org/10.1002/art.39441

    Article  CAS  PubMed  Google Scholar 

  8. Wood KJ, Bushell A, Hester J (2012) Regulatory immune cells in transplantation. Nat Rev Immunol 12(6):417–430. https://doi.org/10.1038/nri3227

    Article  CAS  PubMed  Google Scholar 

  9. Bronte V, Brandau S, Chen SH, Colombo MP, Frey AB, Greten TF, Mandruzzato S, Murray PJ, Ochoa A, Ostrand-Rosenberg S, Rodriguez PC, Sica A, Umansky V, Vonderheide RH, Gabrilovich DI (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun 7:12150. https://doi.org/10.1038/ncomms12150

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Pawelec G, Verschoor CP, Ostrand-Rosenberg S (2019) Myeloid-derived suppressor cells: not only in tumor immunity. Front Immunol 10:1099. https://doi.org/10.3389/fimmu.2019.01099

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Cassetta L, Baekkevold ES, Brandau S, Bujko A, Cassatella MA, Dorhoi A, Krieg C, Lin A, Lore K, Marini O, Pollard JW, Roussel M, Scapini P, Umansky V, Adema GJ (2019) Deciphering myeloid-derived suppressor cells: isolation and markers in humans, mice and non-human primates. Cancer Immunol Immunother 68(4):687–697. https://doi.org/10.1007/s00262-019-02302-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Alshetaiwi H, Pervolarakis N, McIntyre LL, Ma D, Nguyen Q, Rath JA, Nee K, Hernandez G, Evans K, Torosian L, Silva A, Walsh C, Kessenbrock K (2020) Defining the emergence of myeloid-derived suppressor cells in breast cancer using single-cell transcriptomics. Sci Immunol 5(44):eaay6017. https://doi.org/10.1126/sciimmunol.aay6017

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Gabrilovich DI (2017) Myeloid-derived suppressor cells. Cancer Immunol Res 5(1):3–8. https://doi.org/10.1158/2326-6066.CIR-16-0297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Chai E, Zhang L, Li C (2019) LOX-1+ PMN-MDSC enhances immune suppression which promotes glioblastoma multiforme progression. Cancer Manag Res 11:7307–7315. https://doi.org/10.2147/CMAR.S210545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Alissafi T, Hatzioannou A, Mintzas K, Barouni RM, Banos A, Sormendi S, Polyzos A, Xilouri M, Wielockx B, Gogas H, Verginis P (2018) Autophagy orchestrates the regulatory program of tumor-associated myeloid-derived suppressor cells. J Clin Invest 128(9):3840–3852. https://doi.org/10.1172/JCI120888

    Article  PubMed  PubMed Central  Google Scholar 

  16. Hatzioannou A, Banos A, Sakelaropoulos T, Fedonidis C, Vidali MS, Kohne M, Handler K, Boon L, Henriques A, Koliaraki V, Georgiadis P, Zoidakis J, Termentzi A, Beyer M, Chavakis T, Boumpas D, Tsirigos A, Verginis P (2020) An intrinsic role of IL-33 in Treg cell-mediated tumor immunoevasion. Nat Immunol 21(1):75–85. https://doi.org/10.1038/s41590-019-0555-2

    Article  CAS  PubMed  Google Scholar 

  17. Alissafi T, Banos A, Boon L, Sparwasser T, Ghigo A, Wing K, Vassilopoulos D, Boumpas D, Chavakis T, Cadwell K, Verginis P (2017) Tregs restrain dendritic cell autophagy to ameliorate autoimmunity. J Clin Invest 127(7):2789–2804. https://doi.org/10.1172/JCI92079

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Panayotis Verginis .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Papaioannou, A.S., Boumpas, A., Papadopoulou, M., Hatzioannou, A., Alissafi, T., Verginis, P. (2021). Measuring Suppressive Activity and Autophagy in Myeloid-Derived Suppressor Cells. In: Brandau, S., Dorhoi, A. (eds) Myeloid-Derived Suppressor Cells. Methods in Molecular Biology, vol 2236. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1060-2_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1060-2_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1059-6

  • Online ISBN: 978-1-0716-1060-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics