High-throughput identification of antigen-specific TCRs by TCR gene capture

Carsten Linnemann, Bianca Heemskerk, Pia Kvistborg, Roelof J. C. Kluin, Dmitriy A. Bolotin, Xiaojing Chen, Kaspar Bresser, Marja Nieuwland, Remko Schotte, Samira Michels, Raquel Gomez-Eerland, Lorenz Jahn, Pleun Hombrink, Nicolas Legrand, Chengyi Jenny Shu, Ilgar Z. Mamedov, Arno Velds, Christian U. Blank, John B. A. G. Haanen, Maria A. TurchaninovaRon M. Kerkhoven, Hergen Spits, Sine Reker Hadrup, Mirjam H. M. Heemskerk, Thomas Blankenstein, Dmitriy M. Chudakov, Gavin M. Bendle, Ton N. M. Schumacher

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The transfer of T cell receptor (TCR) genes into patient T cells is a promising approach for the treatment of both viral infections and cancer. Although efficient methods exist to identify antibodies for the treatment of these diseases, comparable strategies to identify TCRs have been lacking. We have developed a high-throughput DNA-based strategy to identify TCR sequences by the capture and sequencing of genomic DNA fragments encoding the TCR genes. We establish the value of this approach by assembling a large library of cancer germline tumor antigen-reactive TCRs. Furthermore, by exploiting the quantitative nature of TCR gene capture, we show the feasibility of identifying antigen-specific TCRs in oligoclonal T cell populations from either human material or TCR-humanized mice. Finally, we demonstrate the ability to identify tumor-reactive TCRs within intratumoral T cell subsets without knowledge of antigen specificities, which may be the first step toward the development of autologous TCR gene therapy to target patient-specific neoantigens in human cancer.
Original languageEnglish
JournalNature Medicine
Volume19
Issue number11
Pages (from-to)1534-1541
ISSN1078-8956
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • Biochemistry, Genetics and Molecular Biology (all)
  • Medicine (all)
  • gamma interferon
  • genomic DNA
  • T lymphocyte receptor alpha chain
  • T lymphocyte receptor beta chain
  • amino acid sequence
  • animal experiment
  • antigen specificity
  • article
  • controlled study
  • cytokine production
  • gene locus
  • gene therapy
  • gene transfer
  • high throughput sequencing
  • human
  • human cell
  • lymphocyte subpopulation
  • mouse
  • nonhuman
  • priority journal
  • protein analysis
  • T lymphocyte receptor gene
  • T lymphocyte subpopulation
  • Animals
  • Antigens, Neoplasm
  • Gene Library
  • Genes, T-Cell Receptor
  • Genetic Therapy
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Mice
  • Neoplasms
  • Receptors, Antigen, T-Cell
  • T-Lymphocytes
  • Mus
  • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
  • Rodentia Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Mammals, Nonhuman Vertebrates, Nonhuman Mammals, Rodents, Vertebrates) - Muridae [86375] mouse common
  • human TCR gene [Hominidae] human T cell receptor gene expression
  • antigen-specific T cell receptor antigen-specific TCR
  • genomic DNA fragments
  • T cell receptor TCR
  • 02506, Cytology - Animal
  • 02508, Cytology - Human
  • 03502, Genetics - General
  • 03506, Genetics - Animal
  • 03508, Genetics - Human
  • 15002, Blood - Blood and lymph studies
  • 15004, Blood - Blood cell studies
  • 34502, Immunology - General and methods
  • 34508, Immunology - Immunopathology, tissue immunology
  • Immune System
  • Methods and Techniques
  • Molecular Genetics
  • autoimmune disease Autoimmune Diseases (MeSH) immune system disease
  • Biochemistry and Molecular Biophysics
  • Chemical Coordination and Homeostasis
  • T cell immune system, blood and lymphatics
  • high-throughput DNA-based method laboratory techniques, genetic techniques
  • BIOCHEMISTRY
  • CELL
  • MEDICINE,
  • T-CELL-RECEPTORS
  • CANCER REGRESSION
  • MONOCLONAL-ANTIBODIES
  • ADOPTIVE TRANSFER
  • MELANOMA PATIENTS
  • IMMUNE-SYSTEM
  • IN-VIVO
  • LYMPHOCYTES
  • THERAPY
  • MICE
  • HIGH throughput screening (Drug development)

Fingerprint Dive into the research topics of 'High-throughput identification of antigen-specific TCRs by TCR gene capture'. Together they form a unique fingerprint.

Cite this