title: Generation of activated and antigen-specific T cells with cytotoxic activity after co-culture with dendritic cells.
abstract: Co-culturing of immunological effector cells with antigen-pulsed DC leads to an increase of cytotoxic activity against antigen-expressing tumour cells. Using this approach, we could detect up to 2.8% antigen-specific CTLs after co-culture with antigen-pulsed DC. However, the required high effector cell numbers remain a major obstacle in immunotherapy. In this study, we show an approach for generating activated and antigen-specific effector cells that enables us to decrease effector to target cell ratios. We used an interferon-gamma secretion assay to enrich activated effector cells after co-culture with antigen-pulsed dendritic cells (DC). Purified immunological effector cells lysed 58.3% of antigen-expressing tumour cells at an effector to target ratio of 1:1. Furthermore, using MHC-IgG complexes, we enriched effector cells expressing antigen-specific T-cell receptor after co-culture with DC. Performing ELISpot, flow cytometry and TCR analysis, we could show a significant increase of activated and specific TCR-expressing effector cells after co-culture with DC.
authors.affiliation: Department of Internal Medicine I, Rheinische Friedrich-Wilhelms-University, Bonn, Germany.
authors.forename: Angela
authors.initials: A
authors.lastname: Märten
authors.forename: Tim
authors.initials: T
authors.lastname: Greten
authors.forename: Carsten
authors.initials: C
authors.lastname: Ziske
authors.forename: Sabine
authors.initials: S
authors.lastname: Renoth
authors.forename: Björn
authors.initials: B
authors.lastname: Schöttker
authors.forename: Peter
authors.initials: P
authors.lastname: Buttgereit
authors.forename: Frank
authors.initials: F
authors.lastname: Schakowski
authors.forename: Alexander
authors.initials: A
authors.lastname: von Rücker
authors.forename: Tilman
authors.initials: T
authors.lastname: Sauerbruch
authors.forename: Ingo H
authors.initials: IH
authors.lastname: Schmidt-Wolf
mesh_terms: D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology*; D002272:Carcinoembryonic Antigen / Q000737:chemistry / Q000276:immunology*; D002478:Cells, Cultured / Q000276:immunology / Q000378:metabolism; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000473:pathology; D003713:Dendritic Cells / Q000276:immunology*; D019281:Dimerization; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D016130:Immunophenotyping; D007371:Interferon-gamma / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D015789:HLA-A2 Antigen; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma