Tumor antigen-specific T cells for immune monitoring of dendritic cell-treated glioblastoma patients

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• 作者：Isabelle Mü ; ller¹ ; ^{isabelle.mueller@uks.eu" class="auth_mail" title="E-mail the corresponding author} ; Dominik Altherr¹ ; Matthias Eyrich² ; Brigitte Flesch³ ; Kim S. Friedmann⁴ ; Ralf Ketter⁵ ; Joachim Oertel⁵ ; Eva C. Schwarz⁴ ; Antje Technau² ; Steffi Urbschat⁵ ; Hermann Eichler¹
• 关键词：CD8+ T cells ; immune monitoring ; GBM peptides ; dextramer staining ; IFN-γ ; cancer immunotherapy
• 刊名：Cytotherapy
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：18
• 期：9
• 页码：1146-1161
• 全文大小：1340 K

文摘

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)-γ fluorescence flow cytometry-based measurements were combined with anti-GBM tumor peptide dextramer staining. IFN-γ 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-γ messenger RNA levels, and intracellular IFN-γ 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-γ 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.