Isolation and characterization of neutrophils with anti-tumor properties

Ronit Vogt Sionov, Simaan Assi, Maya Gershkovitz, Jitka Y. Sagiv, Lola Polyansky, Inbal Mishalian, Zvi G. Fridlender, Zvi Granot*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

Neutrophils, the most abundant of all white blood cells in the human circulation, play an important role in the host defense against invading microorganisms. In addition, neutrophils play a central role in the immune surveillance of tumor cells. They have the ability to recognize tumor cells and induce tumor cell death either through a cell contact-dependent mechanism involving hydrogen peroxide or through antibodydependent cell-mediated cytotoxicity (ADCC). Neutrophils with anti-tumor activity can be isolated from peripheral blood of cancer patients and of tumor-bearing mice. These neutrophils are termed tumor-entrained neutrophils (TEN) to distinguish them from neutrophils of healthy subjects or naïve mice that show no significant tumor cytotoxic activity. Compared with other white blood cells, neutrophils show different buoyancy making it feasible to obtain a > 98% pure neutrophil population when subjected to a density gradient. However, in addition to the normal high-density neutrophil population (HDN), in cancer patients, in tumor-bearing mice, as well as under chronic inflammatory conditions, distinct low-density neutrophil populations (LDN) appear in the circulation. LDN co-purify with the mononuclear fraction and can be separated from mononuclear cells using either positive or negative selection strategies. Once the purity of the isolated neutrophils is determined by flow cytometry, they can be used for in vitro and in vivo functional assays. We describe techniques for monitoring the anti-tumor activity of neutrophils, their ability to migrate and to produce reactive oxygen species, as well as monitoring their phagocytic capacity ex vivo. We further describe techniques to label the neutrophils for in vivo tracking, and to determine their anti-metastatic capacity in vivo. All these techniques are essential for understanding how to obtain and characterize neutrophils with anti-tumor function.

Original languageAmerican English
Article numbere52933
JournalJournal of Visualized Experiments
Volume2015
Issue number100
DOIs
StatePublished - 19 Jun 2015

Bibliographical note

Publisher Copyright:
© 2015 Journal of Visualized Experiments.

Keywords

  • Anti-metastatic activity
  • Anti-tumor cytotoxicity
  • BrdU labeling
  • CFSE labeling
  • High-density neutrophils
  • Immunology
  • Issue 100
  • Low-density neutrophils
  • Luciferase assay
  • Lung metastatic seeding assay
  • Neutrophil adoptive transfer
  • Neutrophil depletion
  • Neutrophil isolation
  • Tumor-entrained neutrophils

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