Skip to main content

Isolation and Characterization of Low- vs. High-Density Neutrophils in Cancer

  • Protocol
  • First Online:
The Tumor Microenvironment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1458))

Abstract

Neutrophils are the most abundant of all white blood cells in the human circulation and serve as the first line of defense against microbial infections. Traditionally, neutrophils were viewed as a homogeneous population of myeloid cells. However, in recent years accumulating evidence has suggested that neutrophils are heterogeneous and that distinct neutrophil subsets may play very different roles. Here, we describe the methodology for isolation of high- and low-density neutrophils from the murine and human circulation using a density gradient and antibody based enrichment. We further describe the methodology for functional characterization of these different neutrophil subsets in the context of cancer.

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 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Pekarek LA, Starr BA, Toledano AY, Schreiber H (1995) Inhibition of tumor growth by elimination of granulocytes. J Exp Med 181(1):435–440

    Article  CAS  PubMed  Google Scholar 

  2. Nozawa H, Chiu C, Hanahan D (2006) Infiltrating neutrophils mediate the initial angiogenic switch in a mouse model of multistage carcinogenesis. Proc Natl Acad Sci U S A 103(33):12493–12498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. De Larco JE, Wuertz BR, Furcht LT (2004) The potential role of neutrophils in promoting the metastatic phenotype of tumors releasing interleukin-8. Clin Cancer Res 10(15):4895–4900

    Article  PubMed  Google Scholar 

  4. Youn JI, Nagaraj S, Collazo M, Gabrilovich DI (2008) Subsets of myeloid-derived suppressor cells in tumor-bearing mice. J Immunol 181(8):5791–5802

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Yan HH, Pickup M, Pang Y, Gorska AE, Li Z, Chytil A, Geng Y, Gray JW, Moses HL, Yang L (2010) Gr-1+CD11b+ myeloid cells tip the balance of immune protection to tumor promotion in the premetastatic lung. Cancer Res 70(15):6139–6149

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Granot Z, Henke E, Comen EA, King TA, Norton L, Benezra R (2011) Tumor entrained neutrophils inhibit seeding in the premetastatic lung. Cancer Cell 20(3):300–314

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Lopez-Lago MA, Posner S, Thodima VJ, Molina AM, Motzer RJ, Chaganti RS (2013) Neutrophil chemokines secreted by tumor cells mount a lung antimetastatic response during renal cell carcinoma progression. Oncogene 32(14):1752–1760

    Article  CAS  PubMed  Google Scholar 

  8. Sionov RV, Fridlender ZG, Granot Z (2014) The multifaceted roles neutrophils play in the tumor microenvironment. Cancer Microenviron 8(3):125–158

    Article  PubMed  PubMed Central  Google Scholar 

  9. Galdiero MR, Bonavita E, Barajon I, Garlanda C, Mantovani A, Jaillon S (2013) Tumor associated macrophages and neutrophils in cancer. Immunobiology 218(11):1402–1410

    Article  CAS  PubMed  Google Scholar 

  10. Sagiv JY, Michaeli J, Assi S, Mishalian I, Kisos H, Levy L, Damti P, Lumbroso D, Polyansky L, Sionov RV, Ariel A, Hovav A-H, Henke E, Fridlender ZG, Granot Z (2015) Phenotypic diversity and plasticity in circulating neutrophil subpopulations in cancer. Cell Rep 10(4):562–573

    Article  CAS  PubMed  Google Scholar 

  11. Brandau S, Trellakis S, Bruderek K, Schmaltz D, Steller G, Elian M, Suttmann H, Schenck M, Welling J, Zabel P, Lang S (2011) Myeloid-derived suppressor cells in the peripheral blood of cancer patients contain a subset of immature neutrophils with impaired migratory properties. J Leukoc Biol 89(2):311–317

    Article  CAS  PubMed  Google Scholar 

  12. Cloke T, Munder M, Taylor G, Muller I, Kropf P (2012) Characterization of a novel population of low-density granulocytes associated with disease severity in HIV-1 infection. PLoS One 7(11):e48939

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Denny MF, Yalavarthi S, Zhao W, Thacker SG, Anderson M, Sandy AR, McCune WJ, Kaplan MJ (2010) A distinct subset of proinflammatory neutrophils isolated from patients with systemic lupus erythematosus induces vascular damage and synthesizes type I IFNs. J Immunol 184(6):3284–3297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Morisaki T, Goya T, Ishimitsu T, Torisu M (1992) The increase of low density subpopulations and CD10 (CALLA) negative neutrophils in severely infected patients. Surg Today 22(4):322–327

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zvi Granot .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Sagiv, J.Y., Voels, S., Granot, Z. (2016). Isolation and Characterization of Low- vs. High-Density Neutrophils in Cancer. In: Ursini-Siegel, J., Beauchemin, N. (eds) The Tumor Microenvironment. Methods in Molecular Biology, vol 1458. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3801-8_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3801-8_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3799-8

  • Online ISBN: 978-1-4939-3801-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics