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.
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References
Pekarek LA, Starr BA, Toledano AY, Schreiber H (1995) Inhibition of tumor growth by elimination of granulocytes. J Exp Med 181(1):435–440
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
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
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
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
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
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
Sionov RV, Fridlender ZG, Granot Z (2014) The multifaceted roles neutrophils play in the tumor microenvironment. Cancer Microenviron 8(3):125–158
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
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
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
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
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
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
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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
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DOI: https://doi.org/10.1007/978-1-4939-3801-8_13
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3801-8
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