title: How to analyze ex vivo T-cell responses in cancer patients.
abstract: Tumor-associated antigens are present in several malignancies and represent promising targets for immunotherapeutic approaches. Antigen-specific CD8+ T-cells are an important component of immunosurveillance in malignant diseases as they recognize, target, and kill tumor cells. The induction of T-cells reactive against epitopes of tumor-associated antigens is the major immunological aim of several cancer vaccination trials. In recent years, significant progress has been made in developing methods to detect and characterize these antigen-specific T-cells. Here, we review and compare the most important techniques for specific T-cell analyses including cytokine-based functional assays, HLA class I/epitope tetrameric complexes, and PCR-based methods. We emphasize single cell methods and underline the importance of ex vivo methods without prior culturing.
authors.affiliation: Immunogenetics Section, Department of Transfusion Medicine, Clinical Center, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892-1502, USA.
authors.forename: Dirk
authors.initials: D
authors.lastname: Nagorsen
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006650:Histocompatibility Testing; D006801:Humans; D007167:Immunotherapy; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000628:therapy; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000951:Antigens, Neoplasm; D016207:Cytokines; D012333:RNA, Messenger