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Tuesday, September 27, 2016 2:16:39 AM
Tumor antigen–specific T cells for immune monitoring of dendritic cell–treated glioblastoma patients
Isabelle Müllercorrespondence the author, Dominik Altherr, Matthias Eyrich, Brigitte Flesch, Kim S. Friedmann, Ralf Ketter, Joachim Oertel, Eva C. Schwarz, Antje Technau, Steffi Urbschat, Hermann Eichler (Germany)
September 2016
Abstract
Background aims
CD8+ T cells are part of the adaptive immune system and, as such, are responsible for the elimination of tumor cells. Dendritic cells (DC) are professional antigen-presenting cells (APC) that activate CD8+ T cells. Effector CD8+ T cells in turn mediate the active immunotherapeutic response of DC vaccination against the aggressive glioblastoma (GBM). The lack of tumor response assays complicates the assessment of treatment success in GBM patients.
Methods
A novel assay to identify specific cytotoxicity of activated T cells by APC was evaluated. Tumor antigen-pulsed DCs from HLA-A*02-positive GBM patients were cultivated to stimulate autologous cytotoxic T lymphocytes (CTL) over a 12-day culture period. To directly correlate antigen specificity and cytotoxic capacity, intracellular interferon (IFN)-y fluorescence flow cytometry-based measurements were combined with anti-GBM tumor peptide dextramer staining. IFN-y response was quantified by real-time polymerase chain reaction (PCR), and selected GBM genes were compared with healthy human brain cDNA by single specific primer PCR characterization.
Results
Using CTL of GBM patients stimulated with GBM lysate-pulsed DCs increased IFN-y messenger RNA levels, and intracellular IFN-y protein expression was positively correlated with specificity against GBM antigens. Moreover, the GBM peptide-specific CD8+ T-cell response correlated with specific GBM gene expression. Following DC vaccination, GBM patients showed 10-fold higher tumor-specific signals compared with unvaccinated GBM patients.
Discussion
These data indicate that GBM tumor peptide-dextramer staining of CTL in combination with intracellular IFN-y staining may be a useful tool to acquire information on whether a specific tumor antigen has the potential to induce an immune response in vivo.
The Journal of Cell Therapy
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