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EGFR-specific T cell frequencies correlate with EGFR expression in head and neck squamous cell carcinoma.

Schuler PJ, Boeckers P, Engers R, Boelke E, Bas M, Greve J, Dumitru CA, Lehnerdt GF, Ferris RL, Andrade Filho PA, Brandau S, Lang S, Whiteside TL, Hoffmann TK - J Transl Med (2011)

Bottom Line: Patients' results were correlated to EGFR expression obtained by immunohistochemistry in corresponding tumor sections.Proliferation and anti-tumor activity of peptide-specific CTL was demonstrated by in vitro stimulation with dendritic cells pulsed with the peptides.EGFR-specific CTL from cancer patients were expanded ex vivo and produced IFN-γ upon recognition of EGFR+ target cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Universität Duisburg-Essen, Hals-Nasen-Ohrenklinik Essen, Germany. patrick.schuler@uk-essen.de

ABSTRACT

Background: In head and neck squamous cell carcinoma (HNSCC), expression levels of the epidermal growth factor receptor (EGFR) correlate with poor prognosis and decreased survival rates. As the mechanisms responsible for cellular immune response to EGFR in vivo remain unclear, the frequency and function of EGFR-specific cytotoxic T cells (CTL) was determined in HNSCC patients.

Methods: The frequency of CTL specific for the HLA-A2.1-restricted EGFR-derived YLN peptide (YLNTVQPTCV) and KLF peptide (KLFGTSGQKT) was determined in 16 HLA-A2.1+ HNSCC patients and 16 healthy HLA-A2.1+ individuals (NC) by multicolor flow cytometry. Patients' results were correlated to EGFR expression obtained by immunohistochemistry in corresponding tumor sections. Proliferation and anti-tumor activity of peptide-specific CTL was demonstrated by in vitro stimulation with dendritic cells pulsed with the peptides.

Results: Frequency of EGFR-specific CTL correlated significantly with EGFR expression in tumor sections (p = 0.02, r2 = 0.6). Patients with elevated EGFR scores (> 7) had a significantly higher frequency of EGFR-specific CTL than NC and patients with low EGFR scores (< 7). EGFR-specific CTL from cancer patients were expanded ex vivo and produced IFN-γ upon recognition of EGFR+ target cells.

Conclusion: EGFR expressed on HNSCC cells induces a specific immune response in vivo. Strategies for expansion of EGFR-specific CTL may be important for future immunotherapy of HNSCC patients.

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Related in: MedlinePlus

Binding capacity of EGFR-specific peptides to HLA-A2 complex on the surface of T2 cells. The FLU peptide showed the best binding capacity to surface HLA-A2 compared to the two EGFR-specific peptides KLF and YLN. In all peptides expression of surface HLA-A2 complex was dose-dependent.
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Figure 1: Binding capacity of EGFR-specific peptides to HLA-A2 complex on the surface of T2 cells. The FLU peptide showed the best binding capacity to surface HLA-A2 compared to the two EGFR-specific peptides KLF and YLN. In all peptides expression of surface HLA-A2 complex was dose-dependent.

Mentions: The ability of the EGFR-specific peptides (KLF, YLN) to stabilize the HLA-A2 complex on the surface of TAP-deficient T2 cells in relation to the FLU peptide is shown in Figure 1. In the T2-assay, stabilisation of the HLA-A2 complex correlates well with the binding capacity of the specific peptide to HLA-A2, which validates the peptide for recognition of EGFR-specific CTL. In this assay, the KLF peptide had a higher binding capacity (79% of FLU) than the YLN peptide (70% of FLU). For all three peptides, expression of surface HLA-A2 decreased in correlation to the peptide concentration. Additionally, the sequences of both EGFR-peptides, as well as the control peptides (FLU and HIV) were entered into the web-based program for peptide binding prediction from NIH (http://www-bimas.cit.nih.gov/molbio/). The scores for the EGFR-peptides YLN (320) and KLF (96) were higher than for HIV (39) but lower than for FLU (550). The predicted binding ability was in accordance to our present results, in which the frequency of EGFR-specific T cells was slightly higher for the YLN peptide compared to the KLF-peptide. For comparison, we also entered a peptide from the HPV1a-protein (ILSRFKDTA) into the binding prediction program, which had been described to have a low affinity to HLA-A2 [15]. The score for the HPV1a-protein was 15.


EGFR-specific T cell frequencies correlate with EGFR expression in head and neck squamous cell carcinoma.

Schuler PJ, Boeckers P, Engers R, Boelke E, Bas M, Greve J, Dumitru CA, Lehnerdt GF, Ferris RL, Andrade Filho PA, Brandau S, Lang S, Whiteside TL, Hoffmann TK - J Transl Med (2011)

Binding capacity of EGFR-specific peptides to HLA-A2 complex on the surface of T2 cells. The FLU peptide showed the best binding capacity to surface HLA-A2 compared to the two EGFR-specific peptides KLF and YLN. In all peptides expression of surface HLA-A2 complex was dose-dependent.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198929&req=5

Figure 1: Binding capacity of EGFR-specific peptides to HLA-A2 complex on the surface of T2 cells. The FLU peptide showed the best binding capacity to surface HLA-A2 compared to the two EGFR-specific peptides KLF and YLN. In all peptides expression of surface HLA-A2 complex was dose-dependent.
Mentions: The ability of the EGFR-specific peptides (KLF, YLN) to stabilize the HLA-A2 complex on the surface of TAP-deficient T2 cells in relation to the FLU peptide is shown in Figure 1. In the T2-assay, stabilisation of the HLA-A2 complex correlates well with the binding capacity of the specific peptide to HLA-A2, which validates the peptide for recognition of EGFR-specific CTL. In this assay, the KLF peptide had a higher binding capacity (79% of FLU) than the YLN peptide (70% of FLU). For all three peptides, expression of surface HLA-A2 decreased in correlation to the peptide concentration. Additionally, the sequences of both EGFR-peptides, as well as the control peptides (FLU and HIV) were entered into the web-based program for peptide binding prediction from NIH (http://www-bimas.cit.nih.gov/molbio/). The scores for the EGFR-peptides YLN (320) and KLF (96) were higher than for HIV (39) but lower than for FLU (550). The predicted binding ability was in accordance to our present results, in which the frequency of EGFR-specific T cells was slightly higher for the YLN peptide compared to the KLF-peptide. For comparison, we also entered a peptide from the HPV1a-protein (ILSRFKDTA) into the binding prediction program, which had been described to have a low affinity to HLA-A2 [15]. The score for the HPV1a-protein was 15.

Bottom Line: Patients' results were correlated to EGFR expression obtained by immunohistochemistry in corresponding tumor sections.Proliferation and anti-tumor activity of peptide-specific CTL was demonstrated by in vitro stimulation with dendritic cells pulsed with the peptides.EGFR-specific CTL from cancer patients were expanded ex vivo and produced IFN-γ upon recognition of EGFR+ target cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Universität Duisburg-Essen, Hals-Nasen-Ohrenklinik Essen, Germany. patrick.schuler@uk-essen.de

ABSTRACT

Background: In head and neck squamous cell carcinoma (HNSCC), expression levels of the epidermal growth factor receptor (EGFR) correlate with poor prognosis and decreased survival rates. As the mechanisms responsible for cellular immune response to EGFR in vivo remain unclear, the frequency and function of EGFR-specific cytotoxic T cells (CTL) was determined in HNSCC patients.

Methods: The frequency of CTL specific for the HLA-A2.1-restricted EGFR-derived YLN peptide (YLNTVQPTCV) and KLF peptide (KLFGTSGQKT) was determined in 16 HLA-A2.1+ HNSCC patients and 16 healthy HLA-A2.1+ individuals (NC) by multicolor flow cytometry. Patients' results were correlated to EGFR expression obtained by immunohistochemistry in corresponding tumor sections. Proliferation and anti-tumor activity of peptide-specific CTL was demonstrated by in vitro stimulation with dendritic cells pulsed with the peptides.

Results: Frequency of EGFR-specific CTL correlated significantly with EGFR expression in tumor sections (p = 0.02, r2 = 0.6). Patients with elevated EGFR scores (> 7) had a significantly higher frequency of EGFR-specific CTL than NC and patients with low EGFR scores (< 7). EGFR-specific CTL from cancer patients were expanded ex vivo and produced IFN-γ upon recognition of EGFR+ target cells.

Conclusion: EGFR expressed on HNSCC cells induces a specific immune response in vivo. Strategies for expansion of EGFR-specific CTL may be important for future immunotherapy of HNSCC patients.

Show MeSH
Related in: MedlinePlus