Tsyn-seq: a T cell synapse–based antigen identification platform

Yimei Jin, Takahiko Miyama, Alexandria Brown, Tomo Hayase, Xingzhi Song, Anand K. Singh, Licai Huang, Ivonne I. Flores, Lauren K. McDaniel, Israel Glover, Taylor M. Halsey, Rishika Prasad, Valerie Chapa, Saira Ahmed, Jianhua Zhang, Kunal Rai, Christine B. Peterson, Gregory Lizee, Jennifer Karmouch, Eiko HayaseJeffrey J. Molldrem, Chia Chi Chang, Wen Bin Tsai, Robert R. Jenq

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Tools for genome-wide rapid identification of peptide–major histocompatibility complex targets of T-cell receptors (TCRs) are not yet universally available. We present a new antigen screening method, the T-synapse (Tsyn) reporter system, which includes antigen-presenting cells (APCs) 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.

Original languageEnglish (US)
JournalCancer Immunology Research
Volume12
Issue number5
DOIs
StatePublished - May 1 2024

Keywords

  • T-cell receptor
  • antigen
  • epitope
  • immune synapse
  • screen

ASJC Scopus subject areas

  • General Medicine

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