Massively parallel interrogation and mining of natively paired human TCRαβ repertoires

Research output: Contribution to journalArticlepeer-review

  • External authors:
  • Matthew J. Spindler
  • Ayla L. Nelson
  • Ellen K. Wagner
  • Natasha Oppermans
  • John S. Bridgeman
  • James M. Heather
  • Adam S. Adler
  • Michael A. Asensio
  • Robert C. Edgar
  • Yoong Wearn Lim
  • Everett H. Meyer
  • Mark Cobbold
  • David S. Johnson

Abstract

T cells engineered to express antigen-specific T cell receptors (TCRs) are potent therapies for viral infections and cancer. However, efficient identification of clinical candidate TCRs is complicated by the size and complexity of T cell repertoires and the challenges of working with primary T cells. Here we present a high-throughput method to identify TCRs with high functional avidity from diverse human T cell repertoires. The approach used massively parallel microfluidics to generate libraries of natively paired, full-length TCRαβ clones, from millions of primary T cells, which were then expressed in Jurkat cells. The TCRαβ–Jurkat libraries enabled repeated screening and panning for antigen-reactive TCRs using peptide major histocompatibility complex binding and cellular activation. We captured more than 2.9 million natively paired TCRαβ clonotypes from six healthy human donors and identified rare (<0.001% frequency) viral-antigen-reactive TCRs. We also mined a tumor-infiltrating lymphocyte sample from a patient with melanoma and identified several tumor-specific TCRs, which, after expression in primary T cells, led to tumor cell killing.

Bibliographical metadata

Original languageEnglish
Pages (from-to)609-619
Number of pages11
JournalNature biotechnology
Volume38
Issue number5
DOIs
Publication statusPublished - 16 Mar 2020

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