Assessment of vaccine-induced CD4 T cell responses to the 119-143 immunodominant region of the tumor-specific antigen NY-ESO-1 using DRB1*0101 tetramers.
OBJECTIVE
NY-ESO-1 (ESO), a tumor-specific antigen of the cancer/testis group, is presently viewed as an important model antigen for the development of generic anticancer vaccines. The ESO(119-143) region is immunodominant following immunization with a recombinant ESO vaccine. In this study, we generated DRB1*0101/ESO(119-143) tetramers and used them to assess CD4 T-cell responses in vaccinated patients expressing DRB1*0101 (DR1).


METHODS
We generated tetramers of DRB1*0101 incorporating peptide ESO(119-143) using a previously described strategy. We assessed ESO(119-143)-specific CD4 T cells in peptide-stimulated postvaccine cultures using the tetramers. We isolated DR1/ESO(119-143) tetramer(+) cells by cell sorting and characterized them functionally. We assessed vaccine-induced CD4(+) DR1/ESO(119-143) tetramer(+) T cells ex vivo and characterized them phenotypically.


RESULTS
Staining of cultures from vaccinated patients with DR1/ESO(119-143) tetramers identified vaccine-induced CD4 T cells. Tetramer(+) cells isolated by cell sorting were of T(H)1 type and efficiently recognized full-length ESO. We identified ESO(123-137) as the minimal optimal epitope recognized by DR1-restricted ESO-specific CD4 T cells. By assessing DR1/ESO(119-143) tetramer(+) cells using T cell receptor (TCR) β chain variable region (Vβ)-specific antibodies, we identified several frequently used Vβ. Finally, direct ex vivo staining of patients' CD4 T cells with tetramers allowed the direct quantification and phenotyping of vaccine-induced ESO-specific CD4 T cells.


CONCLUSIONS
The development of DR1/ESO(119-143) tetramers, allowing the direct visualization, isolation, and characterization of ESO-specific CD4 T cells, will be instrumental for the evaluation of spontaneous and vaccine-induced immune responses to this important tumor antigen in DR1-expressing patients.
Ayyoub
Maha
M
Institut National de Santé et de Recherche Médicale, Unité 892, CLCC René Gauducheau, Saint Herblain, France. Maha.Ayyoub@univ-nantes.fr
Pignon
Pascale
P
Dojcinovic
Danijel
D
Raimbaud
Isabelle
I
Old
Lloyd J
LJ
Luescher
Immanuel
I
Valmori
Danila
D
D000595:Amino Acid Sequence; D000818:Animals; D000937:Antigen-Antibody Reactions / Q000276:immunology; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D019496:Cancer Vaccines / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002478:Cells, Cultured; D002986:Clinical Trials as Topic; D015234:HLA-A Antigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D059811:HLA-DRB1 Chains; D006801:Humans; D007111:Immunity, Cellular / Q000187:drug effects / Q000276:immunology; D016056:Immunodominant Epitopes / Q000276:immunology*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D009363:Neoplasm Proteins / Q000737:chemistry / Q000276:immunology; D055503:Protein Multimerization / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D014611:Vaccination / Q000379:methods; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D054928:Validation Studies as Topic
D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D015234:HLA-A Antigens; D059811:HLA-DRB1 Chains; C487961:HLA-DRB1*01:01 antigen; D016056:Immunodominant Epitopes; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic