Tsyn-Seq: a T-cell Synapse-Based Antigen Identification Platform

Cancer Immunol Res. 2024 May 2;12(5):530-543. doi: 10.1158/2326-6066.CIR-23-0467.

Abstract

Tools for genome-wide rapid identification of peptide-major histocompatibility complex targets of T-cell receptors (TCR) are not yet universally available. We present a new antigen screening method, the T-synapse (Tsyn) reporter system, which includes antigen-presenting cells (APC) with a Fas-inducible NF-κB reporter and T cells with a nuclear factor of activated T cells (NFAT) reporter. To functionally screen for target antigens from a cDNA library, productively interacting T cell-APC aggregates were detected by dual-reporter activity and enriched by flow sorting followed by antigen identification quantified by deep sequencing (Tsyn-seq). When applied to a previously characterized TCR specific for the E7 antigen derived from human papillomavirus type 16 (HPV16), Tsyn-seq successfully enriched the correct cognate antigen from a cDNA library derived from an HPV16-positive cervical cancer cell line. Tsyn-seq provides a method for rapidly identifying antigens recognized by TCRs of interest from a tumor cDNA library. See related Spotlight by Makani and Joglekar, p. 515.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Antigen-Presenting Cells / immunology
  • Cell Line, Tumor
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Human papillomavirus 16 / genetics
  • Human papillomavirus 16 / immunology
  • Humans
  • Immunological Synapses* / immunology
  • NFATC Transcription Factors / immunology
  • NFATC Transcription Factors / metabolism
  • Papillomavirus E7 Proteins / genetics
  • Papillomavirus E7 Proteins / immunology
  • Receptors, Antigen, T-Cell* / genetics
  • Receptors, Antigen, T-Cell* / immunology
  • T-Lymphocytes* / immunology

Substances

  • oncogene protein E7, Human papillomavirus type 16