title: A new cloning and expression system yields and validates TCRs from blood lymphocytes of patients with cancer within 10 days.
abstract: Antigen-specific T cell therapy, or T cell receptor (TCR) gene therapy, is a promising immunotherapy for infectious diseases and cancers. However, a suitable rapid and direct screening system for antigen-specific TCRs is not available. Here, we report an efficient cloning and functional evaluation system to determine the antigen specificity of TCR cDNAs derived from single antigen-specific human T cells within 10 d. Using this system, we cloned and analyzed 380 Epstein-Barr virus-specific TCRs from ten healthy donors with latent Epstein-Barr virus infection and assessed the activity of cytotoxic T lymphocytes (CTLs) carrying these TCRs against antigenic peptide-bearing target cells. We also used this system to clone tumor antigen-specific TCRs from peptide-vaccinated patients with cancer. We obtained 210 tumor-associated antigen-specific TCRs and demonstrated the cytotoxic activity of CTLs carrying these TCRs against peptide-bearing cells. This system may provide a fast and powerful approach for TCR gene therapy for infectious diseases and cancers.
authors.affiliation: 1] Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan. [2].
authors.forename: Eiji
authors.initials: E
authors.lastname: Kobayashi
authors.forename: Eishiro
authors.initials: E
authors.lastname: Mizukoshi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishi
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Ozawa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hamana
authors.forename: Terumi
authors.initials: T
authors.lastname: Nagai
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Aishun
authors.initials: A
authors.lastname: Jin
authors.forename: Shuichi
authors.initials: S
authors.lastname: Kaneko
authors.forename: Atsushi
authors.initials: A
authors.lastname: Muraguchi
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000276:immunology / Q000628:therapy; D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D019672:Genes, T-Cell Receptor; D019673:Genes, T-Cell Receptor alpha; D019674:Genes, T-Cell Receptor beta; D015316:Genetic Therapy; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D007167:Immunotherapy; D008113:Liver Neoplasms / Q000235:genetics / Q000276:immunology / Q000628:therapy; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000628:therapy; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D000509:alpha-Fetoproteins / Q000235:genetics / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D011948:Receptors, Antigen, T-Cell; D000509:alpha-Fetoproteins