Identification of a new hTERT-derived HLA-A*0201 restricted, naturally processed CTL epitope

Mette Thorn; Mingjun Wang; Henrik Kløverpris; Esben G W Schmidt; Anders Fomsgaard; Lynn Wenandy; Annika Berntsen; Søren Brunak; Soren Buus; Mogens H Claesson

doi:10.1007/s00262-007-0319-y

Identification of a new hTERT-derived HLA-A*0201 restricted, naturally processed CTL epitope

Mette Thorn, Mingjun Wang, Henrik Kløverpris, Esben G W Schmidt, Anders Fomsgaard, Lynn Wenandy, Annika Berntsen, Søren Brunak, Soren Buus, Mogens H Claesson

Ph.d.-studienævnet for Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

20 Citationer (Scopus)

Abstract

Udgivelsesdato: 2007-Nov

Originalsprog	Engelsk
Tidsskrift	Cancer Immunology, Immunotherapy
Vol/bind	56
Udgave nummer	11
Sider (fra-til)	1755-63
Antal sider	8
ISSN	0340-7004
DOI	https://doi.org/10.1007/s00262-007-0319-y
Status	Udgivet - 2007

Bibliografisk note

Keywords: Amino Acid Sequence; Amino Acid Substitution; Animals; Antigen Presentation; Blotting, Western; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Epitopes, T-Lymphocyte; HLA-A Antigens; Humans; Mice; Mice, Transgenic; Molecular Sequence Data; Peptide Fragments; Protein Binding; T-Lymphocytes, Cytotoxic; Telomerase

Adgang til dokumentet

10.1007/s00262-007-0319-y

Citationsformater

@article{2be90bf0ebc911ddbf70000ea68e967b,

title = "Identification of a new hTERT-derived HLA-A*0201 restricted, naturally processed CTL epitope",

abstract = "By the use of a neural network capable of performing quantitative predictions of peptides binding to HLA-A*0201 molecules, we identified a number of nonamer peptides derived from the catalytic subunit of telomerase, human telomerase reverse transcriptase (hTERT). Five nonimmunogenic peptides with measured binding affinities for HLA-A*0201 ranging from 155 to 1,298 nM were modified at the P1, P2 and P9 positions, respectively, to achieve stronger HLA-A*0201 binding. One peptide, mp30-38 (mp30), with an L to V substitution at position 9 was subsequently found to be immunogenic in mp30 immunized HLA-A*0201/H2K(b) or HHD transgenic mice. The T cell reactivity obtained was directed against both the mp30 and against the unmodified p30. Anti-mp30 specific T cells generated in HLA-A*0201 transgenic mice were dependent on TCR-CD8/MHC-I alpha3 binding and therefore not capable of recognizing mp30-pulsed human HLA-A*0201(+) cells or murine HLA-A*0201 transfectants. In order to show reactivity against naturally processed peptide in human tumor cells, an hTERT positive HLA-A*0201 negative colon carcinoma cell line (CCL220) was transfected with an HLA-A*0201/H2K(b) cDNA construct and used as target in ELISPOT and cytotoxicity assays. The data show that T cells from mp30 immunized HHD transgenic mice react specifically against the CCL220 transfectant indicating that p30 is naturally processed. In conclusion, we have identified a new CTL HLA-A*0201 restricted hTERT epitope, which is now, included in an ongoing phase 2 vaccine trial of patients with disseminated cancer.",

author = "Mette Thorn and Mingjun Wang and Henrik Kl{\o}verpris and Schmidt, {Esben G W} and Anders Fomsgaard and Lynn Wenandy and Annika Berntsen and S{\o}ren Brunak and Soren Buus and Claesson, {Mogens H}",

note = "Keywords: Amino Acid Sequence; Amino Acid Substitution; Animals; Antigen Presentation; Blotting, Western; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Epitopes, T-Lymphocyte; HLA-A Antigens; Humans; Mice; Mice, Transgenic; Molecular Sequence Data; Peptide Fragments; Protein Binding; T-Lymphocytes, Cytotoxic; Telomerase",

year = "2007",

doi = "10.1007/s00262-007-0319-y",

language = "English",

volume = "56",

pages = "1755--63",

journal = "Cancer Immunology, Immunotherapy",

issn = "0340-7004",

publisher = "Springer",

number = "11",

}

TY - JOUR

T1 - Identification of a new hTERT-derived HLA-A*0201 restricted, naturally processed CTL epitope

AU - Thorn, Mette

AU - Wang, Mingjun

AU - Kløverpris, Henrik

AU - Schmidt, Esben G W

AU - Fomsgaard, Anders

AU - Wenandy, Lynn

AU - Berntsen, Annika

AU - Brunak, Søren

AU - Buus, Soren

AU - Claesson, Mogens H

N1 - Keywords: Amino Acid Sequence; Amino Acid Substitution; Animals; Antigen Presentation; Blotting, Western; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Epitopes, T-Lymphocyte; HLA-A Antigens; Humans; Mice; Mice, Transgenic; Molecular Sequence Data; Peptide Fragments; Protein Binding; T-Lymphocytes, Cytotoxic; Telomerase

PY - 2007

Y1 - 2007

N2 - By the use of a neural network capable of performing quantitative predictions of peptides binding to HLA-A*0201 molecules, we identified a number of nonamer peptides derived from the catalytic subunit of telomerase, human telomerase reverse transcriptase (hTERT). Five nonimmunogenic peptides with measured binding affinities for HLA-A*0201 ranging from 155 to 1,298 nM were modified at the P1, P2 and P9 positions, respectively, to achieve stronger HLA-A*0201 binding. One peptide, mp30-38 (mp30), with an L to V substitution at position 9 was subsequently found to be immunogenic in mp30 immunized HLA-A*0201/H2K(b) or HHD transgenic mice. The T cell reactivity obtained was directed against both the mp30 and against the unmodified p30. Anti-mp30 specific T cells generated in HLA-A*0201 transgenic mice were dependent on TCR-CD8/MHC-I alpha3 binding and therefore not capable of recognizing mp30-pulsed human HLA-A*0201(+) cells or murine HLA-A*0201 transfectants. In order to show reactivity against naturally processed peptide in human tumor cells, an hTERT positive HLA-A*0201 negative colon carcinoma cell line (CCL220) was transfected with an HLA-A*0201/H2K(b) cDNA construct and used as target in ELISPOT and cytotoxicity assays. The data show that T cells from mp30 immunized HHD transgenic mice react specifically against the CCL220 transfectant indicating that p30 is naturally processed. In conclusion, we have identified a new CTL HLA-A*0201 restricted hTERT epitope, which is now, included in an ongoing phase 2 vaccine trial of patients with disseminated cancer.

AB - By the use of a neural network capable of performing quantitative predictions of peptides binding to HLA-A*0201 molecules, we identified a number of nonamer peptides derived from the catalytic subunit of telomerase, human telomerase reverse transcriptase (hTERT). Five nonimmunogenic peptides with measured binding affinities for HLA-A*0201 ranging from 155 to 1,298 nM were modified at the P1, P2 and P9 positions, respectively, to achieve stronger HLA-A*0201 binding. One peptide, mp30-38 (mp30), with an L to V substitution at position 9 was subsequently found to be immunogenic in mp30 immunized HLA-A*0201/H2K(b) or HHD transgenic mice. The T cell reactivity obtained was directed against both the mp30 and against the unmodified p30. Anti-mp30 specific T cells generated in HLA-A*0201 transgenic mice were dependent on TCR-CD8/MHC-I alpha3 binding and therefore not capable of recognizing mp30-pulsed human HLA-A*0201(+) cells or murine HLA-A*0201 transfectants. In order to show reactivity against naturally processed peptide in human tumor cells, an hTERT positive HLA-A*0201 negative colon carcinoma cell line (CCL220) was transfected with an HLA-A*0201/H2K(b) cDNA construct and used as target in ELISPOT and cytotoxicity assays. The data show that T cells from mp30 immunized HHD transgenic mice react specifically against the CCL220 transfectant indicating that p30 is naturally processed. In conclusion, we have identified a new CTL HLA-A*0201 restricted hTERT epitope, which is now, included in an ongoing phase 2 vaccine trial of patients with disseminated cancer.

U2 - 10.1007/s00262-007-0319-y

DO - 10.1007/s00262-007-0319-y

M3 - Journal article

C2 - 17464507

SN - 0340-7004

VL - 56

SP - 1755

EP - 1763

JO - Cancer Immunology, Immunotherapy

JF - Cancer Immunology, Immunotherapy

IS - 11

ER -