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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
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
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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.
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Date
2023-05-17
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Nature Research
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Keywords
Biotechnology, Molecular biology
Citation
Fahad, A.S., Chung, C.Y., López Acevedo, S.N. et al. Cell activation-based screening of natively paired human T cell receptor repertoires. Sci Rep 13, 8011 (2023). https://doi.org/10.1038/s41598-023-31858-4