title: Isolation and Characterization of Neutrophils with Anti-Tumor Properties.
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 antibody-dependent 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.
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School.
authors.forename: Ronit Vogt
authors.initials: RV
authors.lastname: Sionov
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School.
authors.forename: Simaan
authors.initials: S
authors.lastname: Assi
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School.
authors.forename: Maya
authors.initials: M
authors.lastname: Gershkovitz
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School.
authors.forename: Jitka Y
authors.initials: JY
authors.lastname: Sagiv
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School.
authors.forename: Lola
authors.initials: L
authors.lastname: Polyansky
authors.affiliation: Institute of Pulmonary Medicine, Hadassah-Hebrew University Medical Center.
authors.forename: Inbal
authors.initials: I
authors.lastname: Mishalian
authors.affiliation: Institute of Pulmonary Medicine, Hadassah-Hebrew University Medical Center.
authors.forename: Zvi G
authors.initials: ZG
authors.lastname: Fridlender
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada, Hebrew University Medical School; zvikag@ekmd.huji.ac.il.
authors.forename: Zvi
authors.initials: Z
authors.lastname: Granot
mesh_terms: D000818:Animals; D005260:Female; D006801:Humans; D007157:Immunologic Surveillance; D008325:Mammary Neoplasms, Experimental / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000276:immunology* / Q000473:pathology; D009504:Neutrophils / Q000166:cytology* / Q000276:immunology* / Q000473:pathology