A new cloning and expression system yields and validates TCRs from blood lymphocytes of patients with cancer within 10 days

Nat Med. 2013 Nov;19(11):1542-6. doi: 10.1038/nm.3358. Epub 2013 Oct 13.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Antigens, Neoplasm / metabolism
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / immunology
  • Carcinoma, Hepatocellular / therapy
  • Cloning, Molecular / methods*
  • Gene Expression
  • Genes, T-Cell Receptor alpha
  • Genes, T-Cell Receptor beta
  • Genes, T-Cell Receptor*
  • Genetic Therapy
  • Herpesvirus 4, Human / immunology
  • Humans
  • Immunotherapy
  • Liver Neoplasms / genetics
  • Liver Neoplasms / immunology
  • Liver Neoplasms / therapy
  • Molecular Sequence Data
  • Neoplasms / genetics*
  • Neoplasms / immunology*
  • Neoplasms / therapy
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / metabolism
  • T-Lymphocytes, Cytotoxic / immunology
  • alpha-Fetoproteins / genetics
  • alpha-Fetoproteins / immunology

Substances

  • Antigens, Neoplasm
  • Cancer Vaccines
  • Receptors, Antigen, T-Cell
  • alpha-Fetoproteins

Associated data

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