title: Negative and positive separation techniques for the isolation of antigen-specific CD8(+) T cells from blood and tumor tissue.
abstract: Adoptive transfer of tumor-reactive cytotoxic T cells (CTLs) is a promising therapeutic approach for the treatment of solid tumors. To develop these strategies, it is necessary to isolate specific leukocyte subpopulations from peripheral blood or tumor tissue (referred to as tumor-infiltrating lymphocytes (TIL)) that will be reinfused into the patient after expansion in vitro. The ideal cell isolation approach should be performed rapidly, thereby maximizing the recovery and viability of the purified cells. Here, we describe the negative or the positive separation procedures to isolate CD8(+) T cells from whole blood, from peripheral blood mononuclear cells (PBMCs), or from cancer tissue. Purified CD8(+) cells will be used for different downstream applications such as protein and gene expression profile analysis in order to assess their intrinsic cytotoxic ability.
authors.affiliation: Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Gigante
authors.forename: Sharon Natasha
authors.initials: SN
authors.lastname: Cox
authors.forename: Elena
authors.initials: E
authors.lastname: Ranieri
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology; D002292:Carcinoma, Renal Cell / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D002499:Centrifugation, Density Gradient; D053804:Granzymes / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D007680:Kidney Neoplasms / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D016246:Lymphocytes, Tumor-Infiltrating / Q000166:cytology*; D008863:Microspheres; D054353:Perforin / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D012397:Rosette Formation / Q000379:methods*; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*
chemical_list: D015395:Histocompatibility Antigens Class I; D011948:Receptors, Antigen, T-Cell; D054353:Perforin; D053804:Granzymes