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Identification of a potential human telomerase reverse transcriptase–derived, HLA-A1–restricted cytotoxic T-lymphocyte epitope

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Abstract

The catalytic subunit of human telomerase reverse transcriptase (hTERT) is expressed in the majority of tumor cells of different histological origins as opposed to most normal somatic cells. This implicates hTERT as a widely expressed tumor-associated antigen and an attractive candidate for antigen-specific tumor immunotherapy. T lymphocytes specific for hTERT-derived epitopes have been isolated and shown reactive with hTERT-expressing tumor cells. To further increase the applicability of hTERT as a target antigen for immunotherapy, we set out to identify potential hTERT-derived, HLA-A1–restricted cytotoxic T-lymphocyte (CTL) epitopes. The “reverse immunology” approach, involving computer-assisted epitope prediction, in vitro CTL induction, and tetramer-guided CTL isolation, resulted in specific CTLs against hTERT-derived, HLA-A1–binding peptides. Intermediate- to low-avidity CTLs were induced against the hTERT325-333 peptide and recognized endogenously processed hTERT. Recognition of endogenous hTERT depended on an increase of hTERT expression above normal levels in tumor cells through hTERT transduction, most probably as a result of limited CTL avidity. The altered peptide ligand hTERT699T-707 was designed to increase HLA-A1–binding affinity of the hTERT699-707 peptide and was used to induce CTLs. However, these CTLs poorly cross-recognized native hTERT699-707 and failed to recognize endogenously processed hTERT. In conclusion, our study has identified the hTERT325-333 peptide as a potential hTERT-derived epitope that may prove useful for induction and monitoring of hTERT-specific, HLA-A1–restricted CTL responses.

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Abbreviations

APC:

Allophycocyanin

CTL:

Cytotoxic T lymphocytes

DC:

Dendritic cell

FITC:

Fluorescein isothiocyanate

GFP:

Green fluorescent protein

hTERT:

Human telomerase reverse transcriptase

IFN:

Interferon

IL:

Interleukin

PE:

Phycoerythrin

PBMC:

Peripheral blood mononuclear cells

T A1:

HLA-A1 tetramer

TCR:

T-cell receptor

TRAP:

Telomere repeat amplification protocol

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Acknowledgements

The authors would like to thank Dr P. van der Bruggen for providing the HLA-A1 encoding construct, Dr R. Weinberg for providing the human hTERT cDNA, Dr H.J. Bontkes and Dr T.D. de Gruijl for critically reading the manuscript, and the Maurits & Anna de Kock foundation for financial support in the purchase of an HPLC.

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Authors and Affiliations

  1. Department of Pathology, VU University Medical Center, de Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands

    Marco W. J. Schreurs, Esther W. M. Kueter, Kirsten B. J. Scholten, Duco Kramer, Chris J. L. M. Meijer & Erik Hooijberg

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  1. Marco W. J. Schreurs
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  2. Esther W. M. Kueter
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  3. Kirsten B. J. Scholten
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  4. Duco Kramer
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  5. Chris J. L. M. Meijer
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  6. Erik Hooijberg
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Corresponding author

Correspondence to Erik Hooijberg.

Additional information

This study was supported by grants VUMC2001-2503 from the Dutch Cancer Society and 901-10-124 from the Netherlands Organisation for Scientific Research.

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Schreurs, M.W.J., Kueter, E.W.M., Scholten, K.B.J. et al. Identification of a potential human telomerase reverse transcriptase–derived, HLA-A1–restricted cytotoxic T-lymphocyte epitope. Cancer Immunol Immunother 54, 703–712 (2005). https://doi.org/10.1007/s00262-004-0611-z

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  • DOI: https://doi.org/10.1007/s00262-004-0611-z

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