학술논문
Cell activation-based screening of natively paired human T cell receptor repertoires.
Document Type
Article
Author
Fahad, Ahmed S.; Chung, Cheng Yu; López Acevedo, Sheila N.; Boyle, Nicoleen; Madan, Bharat; Gutiérrez-González, Matías F.; Matus-Nicodemos, Rodrigo; Laflin, Amy D.; Ladi, Rukmini R.; Zhou, John; Wolfe, Jacy; Llewellyn-Lacey, Sian; Koup, Richard A.; Douek, Daniel C.; Balfour Jr., Henry H.; Price, David A.; DeKosky, Brandon J.
Source
Subject
*T cell receptors
*MAJOR histocompatibility complex
*PEPTIDES
*MOLECULAR cloning
*T cells
*VIRUS diseases
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Language
ISSN
2045-2322
Abstract
Adoptive immune therapies based on the transfer of antigen-specific T cells have been used successfully to treat various cancers and viral infections, but improved techniques are needed to identify optimally protective human T cell receptors (TCRs). Here we present a high-throughput approach to the identification of natively paired human TCRα and TCRβ (TCRα:β) genes encoding heterodimeric TCRs that recognize specific peptide antigens bound to major histocompatibility complex molecules (pMHCs). We first captured and cloned TCRα:β genes from individual cells, ensuring fidelity using a suppression PCR. We then screened TCRα:β libraries expressed in an immortalized cell line using peptide-pulsed antigen-presenting cells and sequenced activated clones to identify the cognate TCRs. Our results validated an experimental pipeline that allows large-scale repertoire datasets to be annotated with functional specificity information, facilitating the discovery of therapeutically relevant TCRs. [ABSTRACT FROM AUTHOR]