Immortalization and functional screening of natively paired human T cell receptor repertoires
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jan 01, 2022
- Source ID
- 10.1093/protein/gzab034
Entities
People
- Ahmed S. Fahad
- Amy D Laflin
- Bharat Madan
- Brandon J DeKosky
- Cheng-yu Chung
- Daniel C. Douek
- David A. Price
- Henry H Balfour Jr
- Jacy Wolfe
- John Zhou
- Matias F Gutiérrez-gonzález
- Nicoleen Boyle
- Richard A. Koup
- Rodrigo Matus-nicodemos
- Rukmini R Ladi
- Sheila N Lopez Acevedo
- Sian Llewellyn-lacey
Organizations
- Cardiff University
- Massachusetts General Hospital
- Massachusetts Institute of Technology
- National Institutes of Health
- Richard M. Schulze Family Foundation
- United States Department of Defense
- University Hospital of Wales
- University of Kansas
- University of Minnesota
- Wellcome