title: Production of NY-ESO-1 peptide/DRB1*08:03 tetramers and ex vivo detection of CD4 T-cell responses in vaccinated cancer patients.
abstract: We established CD4 T-cell clones, Mz-1B7, and Ue-21, which recognized the NY-ESO-1 121-138 peptide from peripheral blood mononuclear cells (PBMCs) of an esophageal cancer patient, E-2, immunized with an NY-ESO-1 protein and determined the NY-ESO-1 minimal epitopes. Minimal peptides recognized by Mz-1B7 and Ue-21 were NY-ESO-1 125-134 and 124-134, respectively, both in restriction to DRB1*08:03. Using a longer peptide, 122-135, and five other related peptides, including either of the minimal epitopes recognized by the CD4 T-cell clones, we investigated the free peptide/DR recognition on autologous EBV-B cells as APC and peptide/DR tetramer binding. The results showed a discrepancy between them. The tetramers with several peptides recognized by either Mz-1B7 or the Ue-21 CD4 T-cell clone did not bind to the respective clone. On the other hand, unexpected binding of the tetramer with the peptide not recognized by CD4 T-cells was observed. The clone Mz-1B7 did not recognize the free peptide 122-135 on APC, but the peptide 122-135/DRB1*08:03 tetramer bound to the TCR on those cells. The failure of tetramer production and the unexpected tetramer binding could be due to a subtly modified structure of the peptide/DR tetramer from the structure of the free peptide/DR molecule. We also demonstrated that the NY-ESO-1 123-135/DRB1*08:03 tetramer detected ex vivo CD4 T-cell responses in PBMCs from patients after NY-ESO-1 vaccination in immunomonitoring.
authors.affiliation: Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan; Department of Respiratory Medicine, Kawasaki Medical School, Okayama, Japan.
authors.forename: Yu
authors.initials: Y
authors.lastname: Mizote
authors.affiliation: Faculty of Health and Welfare, Kawasaki University of Medical Welfare, Kurashiki 701-0193, Okayama, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Uenaka
authors.affiliation: Department of Respiratory Medicine, Kawasaki Medical School, Okayama, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Isobe
authors.affiliation: Department of Surgery, Osaka University Graduate School of Medicine, Osaka, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Wada
authors.affiliation: Department of Immunotherapeutics, University of Tokyo Hospital, Tokyo, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Kakimi
authors.affiliation: Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Saika
authors.affiliation: Medical & Biological Laboratories, Nagano, Japan.
authors.forename: Shoichi
authors.initials: S
authors.lastname: Kita
authors.affiliation: Medical & Biological Laboratories, Nagano, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Koide
authors.affiliation: Department of Respiratory Medicine, Kawasaki Medical School, Okayama, Japan.
authors.forename: Mikio
authors.initials: M
authors.lastname: Oka
authors.affiliation: Faculty of Health and Welfare, Kawasaki University of Medical Welfare, Kurashiki 701-0193, Okayama, Japan. Electronic address: nakayama@mw.kawasaki-m.ac.jp.
authors.forename: Eiichi
authors.initials: E
authors.lastname: Nakayama
mesh_terms: D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology*; D002460:Cell Line; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D004938:Esophageal Neoplasms / Q000628:therapy; D059806:HLA-DR beta-Chains / Q000276:immunology*; D006801:Humans; D008175:Lung Neoplasms / Q000628:therapy; D008297:Male; D008565:Membrane Proteins / Q000276:immunology*; D008969:Molecular Sequence Data; D011471:Prostatic Neoplasms / Q000628:therapy; D022223:Vaccines, Subunit / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D018984:Epitopes, T-Lymphocyte; D059806:HLA-DR beta-Chains; D008565:Membrane Proteins; D022223:Vaccines, Subunit
keywords: APC; CD4 T-cell; CHP-NY-ESO-1; Fmoc; HD; MFI; MHC class II tetramer; N-(9-fluorenyl)-methoxycarbonyl; NY-ESO-1; OLP; PBMC; Tumor immunology; antigen-presenting cell; complex of cholesterol-bearing hydrophobized pullulan and NY-ESO-1 whole protein; healthy donor; mean fluorescence intensity; overlapping peptide; peripheral blood mononuclear cell