A polyclonal anti-vaccine CD4 T cell response detected with HLA-DP4 multimers in a melanoma patient vaccinated with MAGE-3.DP4-peptide-pulsed dendritic cells

Eur J Immunol. 2005 Apr;35(4):1066-75. doi: 10.1002/eji.200425847.


During the last few years, HLA class I tetramers have been successfully used to demonstrate anti-vaccine CD8 CTL proliferation in cancer patients vaccinated with tumor antigens. Frequencies of CTL as low as 10(-6) among CD8 cells were observed even in patients showing tumor regression. Little is known about the role of tumor-antigen-specific CD4 T cells in the context of these anti-vaccine responses. Therefore, we developed a very sensitive approach using fluorescent class-II-peptide multimers to detect antigen-specific CD4 T cells in vaccinated cancer patients. We produced HLA-DP4 multimers loaded with the MAGE-3(243-258) peptide and used them to stain ex vivo PBL from melanoma patients injected with dendritic cells pulsed with several class I and class II tumor antigenic peptides, including the MAGE-3(243-258) peptide. The multimer(+) CD4 T cells were sorted and amplified in clonal conditions; specificity was assessed by their ability to secrete IFN-gamma upon contact with the MAGE-3 antigen. We detected frequencies of about 1x10(-6) anti-MAGE-3.DP4 cells among CD4 cells. A detailed analysis of one patient showed an anti-MAGE-3.DP4 CD4 T cell amplification of at least 3000-fold upon immunization. TCR analysis of the clones from this patient demonstrated a polyclonal response against the MAGE-3 peptide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neoplasm / immunology
  • Antigen Presentation
  • Antigens, Neoplasm / immunology*
  • Antigens, Neoplasm / metabolism
  • Biotin
  • CD4-Positive T-Lymphocytes / immunology*
  • Cancer Vaccines*
  • Dendritic Cells / immunology*
  • Dendritic Cells / transplantation
  • Epitopes, T-Lymphocyte / immunology
  • HLA-DP Antigens / immunology*
  • HLA-DP Antigens / metabolism
  • HLA-DP beta-Chains
  • Humans
  • Immunotherapy, Adoptive
  • Melanoma / immunology*
  • Melanoma / therapy
  • Neoplasm Proteins / immunology*
  • Neoplasm Proteins / metabolism
  • Peptide Fragments / immunology*
  • Peptide Fragments / metabolism
  • Vaccination


  • Antibodies, Neoplasm
  • Antigens, Neoplasm
  • Cancer Vaccines
  • Epitopes, T-Lymphocyte
  • HLA-DP Antigens
  • HLA-DP beta-Chains
  • HLA-DPw4 antigen
  • MAGEA3 protein, human
  • Neoplasm Proteins
  • Peptide Fragments
  • Biotin