HLA-A3 restricted mutant ras specific cytotoxic T-lymphocytes induced by vaccination with T-helper epitopes

J Mol Med (Berl). 2003 Jan;81(1):43-50. doi: 10.1007/s00109-002-0390-y. Epub 2002 Nov 13.


Cytotoxic T-lymphocytes are one of the most important elements of the antitumor defense. Stimulation of cytotoxic T-lymphocytes outgrowth after immunization with mutant ras peptides is a desired goal since these cells may kill tumor cells in vivo. In this study we tested responding peripheral mononuclear cells from a patient with pancreatic adenocarcinoma who had received intradermal peptide vaccination with a mixture of 17-mer mutant ras peptides and granulocyte-macrophage colony-stimulating factor as an adjuvant. Responding peripheral T-cells were cloned by limiting dilution and several CD8(+) cytotoxic T-lymphocytes, specific for the K- RAS 12-Cys mutation were obtained. By using a panel of nonamer peptides containing the 12-Cys mutation and covering position 4-21 in the ras molecule, the 9-mer peptide which was actually recognized by the cytotoxic T-lymphocytes could be identified. HLA-A*0302 could be identified as the antigen-presenting molecule, and the amino acid sequence of the T-cell epitope carries the previously identified HLA-A*0302 binding motif. The nonamer peptide was contained within the vaccine peptide originally used for intradermal immunization of the patient. The cytotoxic T-lymphocytes were capable of killing target cells expressing HLA-A*0302 that coexpressed the K- RAS 12-Cys mutation after transfection. These data demonstrate that the peptide used for vaccination (17-mer) is processed and presented in vivo, and that generation of cytotoxic T-lymphocytes by vaccination with T-helper epitopes may be important for further development of specific immunotherapy of cancer patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / immunology
  • Adult
  • Antigen Presentation
  • Antigens, Neoplasm / immunology
  • Cancer Vaccines / immunology
  • Cancer Vaccines / therapeutic use*
  • Epitopes / chemistry
  • Epitopes / immunology
  • Female
  • Genes, ras*
  • HLA-A3 Antigen / immunology*
  • Humans
  • Mutation
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / immunology
  • Pancreatic Neoplasms / therapy
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Peptide Fragments / therapeutic use
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Proto-Oncogene Proteins p21(ras) / immunology
  • T-Lymphocytes, Cytotoxic / immunology*
  • Treatment Outcome
  • Vaccination
  • Vaccines, Synthetic / immunology
  • Vaccines, Synthetic / therapeutic use*


  • Antigens, Neoplasm
  • Cancer Vaccines
  • Epitopes
  • HLA-A3 Antigen
  • Peptide Fragments
  • Vaccines, Synthetic
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)