title: Specific targeting of a naturally presented osteosarcoma antigen, papillomavirus binding factor peptide, using an artificial monoclonal antibody.
abstract: Osteosarcoma is a rare but highly malignant tumor occurring most frequently in adolescents. The prognosis of non-responders to chemotherapy is still poor, and new treatment modalities are needed. To develop peptide-based immunotherapy, we previously identified autologous cytotoxic T lymphocyte-defined osteosarcoma antigen papillomavirus binding factor (PBF) in the context of HLA-B55 and the cytotoxic T lymphocyte epitope (PBF A2.2) presented by HLA-A2. PBF and HLA class I are expressed in ∼90 and 70% of various sarcomas, respectively. However, the expression status of peptide PBF A2.2 presented by HLA-A2 on osteosarcoma cells has remained unknown because it is difficult to generate a specific probe that reacts with the HLA·peptide complex. For detection and qualification of the HLA-A*02:01·PBF A2.2 peptide complex on osteosarcoma cells, we tried to isolate a single chain variable fragment (scFv) antibody directed to the HLA-*A0201·PBF A2.2 complex using a naïve scFv phage display library. As a result, scFv clone D12 with high affinity (KD = 1.53 × 10(-9) M) was isolated. D12 could react with PBF A2.2 peptide-pulsed T2 cells and HLA-A2+PBF+ osteosarcoma cell lines and simultaneously demonstrated that the HLA·peptide complex was expressed on osteosarcoma cells. In conclusion, scFv clone D12 might be useful to select candidate patients for PBF A2.2 peptide-based immunotherapy and develop antibody-based immunotherapy.
authors: From the Department of Pathology, tukahara@sapmed.ac.jp.
authors: Tomohide
authors: T
authors: Tsukahara
authors: Department of Orthopaedic Surgery, Sapporo Medial University School of Medicine, Sapporo 060-8543, Japan.
authors: Makoto
authors: M
authors: Emori
authors: From the Department of Pathology, Department of Orthopaedic Surgery, Sapporo Medial University School of Medicine, Sapporo 060-8543, Japan.
authors: Kenji
authors: K
authors: Murata
authors: M.D./Ph.D. program, and.
authors: Takahisa
authors: T
authors: Hirano
authors: M.D./Ph.D. program, and.
authors: Norihiro
authors: N
authors: Muroi
authors: M.D./Ph.D. program, and.
authors: Masanori
authors: M
authors: Kyono
authors: Ina Laboratory, Medical and Biological Laboratories Company, Limited, Ina 396-0002, Japan.
authors: Shingo
authors: S
authors: Toji
authors: Ina Laboratory, Medical and Biological Laboratories Company, Limited, Ina 396-0002, Japan.
authors: Kazue
authors: K
authors: Watanabe
authors: From the Department of Pathology.
authors: Toshihiko
authors: T
authors: Torigoe
authors: From the Department of Pathology.
authors: Vitaly
authors: V
authors: Kochin
authors: Division of Surgical Pathology, Sapporo Medical University Hospital, Sapporo 060-8543, Japan.
authors: Hiroko
authors: H
authors: Asanuma
authors: Department of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo 060-8543, Japan.
authors: Hiroshi
authors: H
authors: Matsumiya
authors: Department of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo 060-8543, Japan.
authors: Keiji
authors: K
authors: Yamashita
authors: Department of Otolaryngology, Sapporo Medical University School of Medicine, Sapporo 060-8543, Japan.
authors: Tetsuo
authors: T
authors: Himi
authors: Department of Immunology, Frontier Medical Research Center, Sapporo Medical University School of Medicine, Sapporo 060-8556, Japan.
authors: Shingo
authors: S
authors: Ichimiya
authors: Department of Orthopaedic Surgery, Sapporo Medial University School of Medicine, Sapporo 060-8543, Japan.
authors: Takuro
authors: T
authors: Wada
authors: Department of Orthopaedic Surgery, Sapporo Medial University School of Medicine, Sapporo 060-8543, Japan.
authors: Toshihiko
authors: T
authors: Yamashita
authors: Division of Surgical Pathology, Sapporo Medical University Hospital, Sapporo 060-8543, Japan.
authors: Tadashi
authors: T
authors: Hasegawa
authors: From the Department of Pathology.
authors: Noriyuki
authors: N
authors: Sato
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D001859:Bone Neoplasms / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology; D015235:HLA-B Antigens / Q000276:immunology; D006801:Humans; D016233:Immunotherapy, Active; D008969:Molecular Sequence Data; D012516:Osteosarcoma / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D027383:Papillomaviridae / Q000276:immunology; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D015789:HLA-A2 Antigen; D015235:HLA-B Antigens; C082944:HLA-B55 antigen; D019151:Peptide Library; D057127:Single-Chain Antibodies
keywords: Antibody Engineering; Antigen Presentation; HLA-A2; HLA/Peptide Complex; Major Histocompatibility Complex (MHC); PBF; Phage Display; T Cell Receptor (TCR); scFv