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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

Staining for EGFR in representative HNSCC samples. The EGFR score (0-16) in tumor samples was calculated as a product of staining intensity (1-4) multiplied by the percentage of positive cells (0-4). Tumor sample with low EGFR expression and EGFR score 2 (a). Tumor sample with high EGFR expression and EGFR score 12 (b). Homogenous expression of EGFR was found in the membranes and cytoplasm of tumor cells (Mag. × 100).
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Figure 3: Staining for EGFR in representative HNSCC samples. The EGFR score (0-16) in tumor samples was calculated as a product of staining intensity (1-4) multiplied by the percentage of positive cells (0-4). Tumor sample with low EGFR expression and EGFR score 2 (a). Tumor sample with high EGFR expression and EGFR score 12 (b). Homogenous expression of EGFR was found in the membranes and cytoplasm of tumor cells (Mag. × 100).

Mentions: All tumor samples were positive for EGFR, and 5 samples showed an EGFR score of 9 or higher. A homogenous ABC-dye uptake was found in tumor cell membranes and cytoplasm of all tumor samples as seen in Figure 3. This staining pattern conformed to staining patterns obtained in the EGFR-positive cell line, UD-SCC-8, which served as a positive control. In the negative control tissues, EGFR expression was observed only in the basal epithelial layers (not shown). Comparing the EGFR scores with the frequency of EGFR-specific CTL revealed a strong positive correlation for both the YLN-peptide (p = 0.02, r2 = 0.6) and the KLF-peptide (p < 0.005, r2 = 0.8) as seen in Figure 4. A clear cut-off was located between the EGFR scores of 6 and 9. None of the early stage tumors (T1) displayed an EGFR-score above 4. For the other tumors (T2-4) samples could be subdivided into weak or strong EGFR expression.


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)

Staining for EGFR in representative HNSCC samples. The EGFR score (0-16) in tumor samples was calculated as a product of staining intensity (1-4) multiplied by the percentage of positive cells (0-4). Tumor sample with low EGFR expression and EGFR score 2 (a). Tumor sample with high EGFR expression and EGFR score 12 (b). Homogenous expression of EGFR was found in the membranes and cytoplasm of tumor cells (Mag. × 100).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Staining for EGFR in representative HNSCC samples. The EGFR score (0-16) in tumor samples was calculated as a product of staining intensity (1-4) multiplied by the percentage of positive cells (0-4). Tumor sample with low EGFR expression and EGFR score 2 (a). Tumor sample with high EGFR expression and EGFR score 12 (b). Homogenous expression of EGFR was found in the membranes and cytoplasm of tumor cells (Mag. × 100).
Mentions: All tumor samples were positive for EGFR, and 5 samples showed an EGFR score of 9 or higher. A homogenous ABC-dye uptake was found in tumor cell membranes and cytoplasm of all tumor samples as seen in Figure 3. This staining pattern conformed to staining patterns obtained in the EGFR-positive cell line, UD-SCC-8, which served as a positive control. In the negative control tissues, EGFR expression was observed only in the basal epithelial layers (not shown). Comparing the EGFR scores with the frequency of EGFR-specific CTL revealed a strong positive correlation for both the YLN-peptide (p = 0.02, r2 = 0.6) and the KLF-peptide (p < 0.005, r2 = 0.8) as seen in Figure 4. A clear cut-off was located between the EGFR scores of 6 and 9. None of the early stage tumors (T1) displayed an EGFR-score above 4. For the other tumors (T2-4) samples could be subdivided into weak or strong EGFR expression.

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