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: Tomohide Tsukahara, Makoto Emori, Kenji Murata, Takahisa Hirano, Norihiro Muroi, Masanori Kyono, Shingo Toji, Kazue Watanabe, Toshihiko Torigoe, Vitaly Kochin, Hiroko Asanuma, Hiroshi Matsumiya, Keiji Yamashita, Tetsuo Himi, Shingo Ichimiya, Takuro Wada, Toshihiko Yamashita, Tadashi Hasegawa, Noriyuki Sato
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 289 Issue 32 Pg. 22035-47 (Aug 08 2014) ISSN: 1083-351X [Electronic] United States
• PMID: 24962571 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• Antibodies, Monoclonal
• Antigens, Neoplasm
• Cancer Vaccines
• HLA-A2 Antigen
• HLA-B Antigens
• HLA-B55 antigen
• Peptide Library
• Single-Chain Antibodies

Topics

• Amino Acid Sequence
• Antibodies, Monoclonal (genetics, immunology)
• Antigen Presentation
• Antigens, Neoplasm (genetics, immunology)
• Base Sequence
• Bone Neoplasms (genetics, immunology, therapy)
• Cancer Vaccines (genetics, immunology)
• Cell Line, Tumor
• HLA-A2 Antigen (genetics, immunology)
• HLA-B Antigens (immunology)
• Humans
• Immunotherapy, Active
• Molecular Sequence Data
• Osteosarcoma (genetics, immunology, therapy)
• Papillomaviridae (immunology)
• Peptide Library
• Single-Chain Antibodies (genetics, immunology)
• T-Lymphocytes, Cytotoxic (immunology)