Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase

John P. Riley, Steven A. Rosenberg, Maria R. Parkhurst

Research output: Contribution to journalArticle

Abstract

Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80% of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma.

Original languageEnglish (US)
Pages (from-to)212-220
Number of pages9
JournalJournal of Immunotherapy
Volume24
Issue number3
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Melanoma-Specific Antigens
Monophenol Monooxygenase
HLA Antigens
Epitopes
Melanoma
Peptides
Blood Cells
Melanocytes
Normal Distribution
Cytotoxic T-Lymphocytes
Cellular Immunity
Immunotherapy
Cell Culture Techniques
T-Lymphocytes
Antigens

Keywords

  • Cytotoxic T lymphocytes (CTL)
  • Epitope
  • HLA-A2.1
  • Melanoma
  • Tyrosinase

ASJC Scopus subject areas

  • Cancer Research
  • Pharmacology
  • Immunology

Cite this

Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase. / Riley, John P.; Rosenberg, Steven A.; Parkhurst, Maria R.

In: Journal of Immunotherapy, Vol. 24, No. 3, 2001, p. 212-220.

Research output: Contribution to journalArticle

Riley, John P. ; Rosenberg, Steven A. ; Parkhurst, Maria R. / Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase. In: Journal of Immunotherapy. 2001 ; Vol. 24, No. 3. pp. 212-220.
@article{3cf383eba9184c9c9559599c3a7d7901,
title = "Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase",
abstract = "Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80{\%} of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma.",
keywords = "Cytotoxic T lymphocytes (CTL), Epitope, HLA-A2.1, Melanoma, Tyrosinase",
author = "Riley, {John P.} and Rosenberg, {Steven A.} and Parkhurst, {Maria R.}",
year = "2001",
doi = "10.1097/00002371-200105000-00004",
language = "English (US)",
volume = "24",
pages = "212--220",
journal = "Journal of Immunotherapy",
issn = "1524-9557",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase

AU - Riley, John P.

AU - Rosenberg, Steven A.

AU - Parkhurst, Maria R.

PY - 2001

Y1 - 2001

N2 - Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80% of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma.

AB - Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80% of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma.

KW - Cytotoxic T lymphocytes (CTL)

KW - Epitope

KW - HLA-A2.1

KW - Melanoma

KW - Tyrosinase

UR - http://www.scopus.com/inward/record.url?scp=0034997840&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034997840&partnerID=8YFLogxK

U2 - 10.1097/00002371-200105000-00004

DO - 10.1097/00002371-200105000-00004

M3 - Article

C2 - 11394498

AN - SCOPUS:0034997840

VL - 24

SP - 212

EP - 220

JO - Journal of Immunotherapy

JF - Journal of Immunotherapy

SN - 1524-9557

IS - 3

ER -