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

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...

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Bibliographic Details
Published in:Scientific reports 2023-05, Vol.13 (1), p.8011-8011, Article 8011
Main Authors: 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., DeKosky, Brandon J.
Format: Article
Language:English
Subjects:
631/337
631/61
Antigen-presenting cells
Antigens
Cell activation
Chemical engineering
Cloning
Cloning, Molecular
Enzymes
Genes
Genomics
Humanities and Social Sciences
Humans
Lymphocytes
Lymphocytes T
Major histocompatibility complex
Medicine
multidisciplinary
Mutation
Peptides
Peptides - genetics
Polymerase chain reaction
Receptors, Antigen, T-Cell
Receptors, Antigen, T-Cell, alpha-beta - genetics
Science
Science (multidisciplinary)
T cell receptors
T-Lymphocytes
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-31858-4