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

• Published in: Cancer immunology research Vol. 12; no. 5; p. 530
• Main Authors: Jin, Yimei, Miyama, Takahiko, Brown, Alexandria, Hayase, Tomo, Song, Xingzhi, Singh, Anand K, Huang, Licai, Flores, Ivonne I, McDaniel, Lauren K, Glover, Israel, Halsey, Taylor M, Prasad, Rishika, Chapa, Valerie, Ahmed, Saira, Zhang, Jianhua, Rai, Kunal, Peterson, Christine B, Lizee, Gregory, Karmouch, Jennifer, Hayase, Eiko, Molldrem, Jeffrey J, Chang, Chia-Chi, Tsai, Wen-Bin, Jenq, Robert R
• Format: Journal Article
• Language: English
• Published: United States 02.05.2024
• Subjects: Antigen-Presenting Cells - immunology; Cell Line, Tumor; Female; Gene Library; High-Throughput Nucleotide Sequencing; Human papillomavirus 16 - genetics; Human papillomavirus 16 - immunology; Humans; Immunological Synapses - immunology; NF-kappa B - metabolism; 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
• ISSN: 2326-6074
• DOI: 10.1158/2326-6066.CIR-23-0467

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.