Defining T cell receptor repertoires using nanovial-based binding and functional screening

• 成果类型: Article
• 署名作者: Koo, Doyeon; Mao, Zhiyuan; Dimatteo, Robert; Noguchi, Miyako; Tsubamoto, Natalie; McLaughlin, Jami; Tran, Wendy; Lee, Sohyung; Cheng, Donghui; de Rutte, Joseph; Sojo, Giselle Burton; Witte, Owen N.; Di Carlo, Dino
• 署名单位: University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; University of California Los Angeles Medical Center; David Geffen School of Medicine at UCLA; University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; UCLA Jonsson Comprehensive Cancer Center; University of California System; University of California Los Angeles; University of California System; University of California Los Angeles; University of California Los Angeles Medical Center; David Geffen School of Medicine at UCLA; University of California System; University of California Los Angeles
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14446
• DOI: 10.1073/pnas.2320442121
• 发表日期: 2024-04-02
• 关键词: ifn-gamma; tcr; selection; single; cd137

摘要

The ability to selectively bind to antigenic peptides and secrete effector molecules can define rare and low- affinity populations of cells with therapeutic potential in emerging T cell receptor (TCR) immunotherapies. We leverage cavity- containing hydrogel microparticles, called nanovials, each coated with peptide - major histocompatibility complex (pMHC) monomers to isolate antigen- reactive T cells. T cells are captured and activated by pMHCs inducing the secretion of effector molecules including IFN-gamma and granzyme B that are accumulated on nanovials, allowing sorting based on both binding and function. The TCRs of sorted cells on nanovials are sequenced, recovering paired alpha beta- chains using microfluidic emulsion - based single - cell sequencing. By labeling nanovials having different pMHCs with unique oligonucleotide- barcodes and secretions with oligo- barcoded detection antibodies, we could accurately link TCR sequences to specific targets and rank each TCR based on the corresponding cell's secretion level. Using the technique, we identified an expanded repertoire of functional TCRs targeting viral antigens with high specificity and found rare TCRs with activity against cancer- specific splicing- enhanced epitopes.