Immortalization and functional screening of natively paired human T cell receptor repertoires
Ahmed S. Fahad, Cheng-Yu Chung, Sheila N. López Acevedo, Nicoleen Boyle, Bharat Madan, Matías Gutiérrez-González, Rodrigo Matus‐Nicodemos, Amy D. Laflin, Rukmini Ladi, John Zhou, Jacy R. Wolfe, Sian Llewellyn‐Lacey, Richard A. Koup, Daniel C. Douek, Henry H. Balfour, David A. Price, Brandon J. DeKosky
Abstract
Functional analyses of the T cell receptor (TCR) landscape can reveal critical information about protection from disease and molecular responses to vaccines. However, it has proven difficult to combine advanced next-generation sequencing technologies with methods to decode the peptide-major histocompatibility complex (pMHC) specificity of individual TCRs. We developed a new high-throughput approach to enable repertoire-scale functional evaluations of natively paired TCRs. In particular, we leveraged the immortalized nature of physically linked TCRα:β amplicon libraries to analyze binding against multiple recombinant pMHCs on a repertoire scale, and to exemplify the utility of this approach, we also performed affinity-based functional mapping in conjunction with quantitative next-generation sequencing to track antigen-specific TCRs. These data successfully validated a new immortalization and screening platform to facilitate detailed molecular analyses of disease-relevant antigen interactions with human TCRs.