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Cell activation-based screening of natively paired human T cell receptor repertoires

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, Henry H., Price, David A. ORCID: and DeKosky, Brandon J. 2023. Cell activation-based screening of natively paired human T cell receptor repertoires. Scientific Reports 13 , 8011. 10.1038/s41598-023-31858-4

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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.

Item Type: Article
Date Type: Published Online
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Additional Information: License information from Publisher: LICENSE 1: URL:, Type: open-access
Publisher: Nature Research
Date of Acceptance: 20 March 2023
Last Modified: 19 May 2023 06:24

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