Generation of activated and antigen-specific T cells with cytotoxic activity after co-culture with dendritic cells.
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.
Märten
Angela
A
Department of Internal Medicine I, Rheinische Friedrich-Wilhelms-University, Bonn, Germany.
Greten
Tim
T
Ziske
Carsten
C
Renoth
Sabine
S
Schöttker
Björn
B
Buttgereit
Peter
P
Schakowski
Frank
F
von Rücker
Alexander
A
Sauerbruch
Tilman
T
Schmidt-Wolf
Ingo H
IH
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
D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D015789:HLA-A2 Antigen; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma