Author ORCID Identifier
Date of Graduation
8-2024
Document Type
Dissertation (PhD)
Program Affiliation
Immunology
Degree Name
Doctor of Philosophy (PhD)
Advisor/Committee Chair
Robert Jenq
Committee Member
Stephanie S. Watowich
Committee Member
Gregory A. Lizee
Committee Member
Linghua Wang
Committee Member
Matthew M. Gubin
Committee Member
Nadim J. Ajami
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.
Recommended Citation
Jin, Yimei, "Tsyn-Seq: A T Cell Synapse-Based Antigen Identification Platform" (2024). Dissertations & Theses (Open Access). 1385.
https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1385
Keywords
T cell