Spindler, M. J. et al. Nat. Biotechnol. https://doi.org/10.1038/s41587-020-0438-y (2020).
T cell receptors (TCRs) consist of α and β chains, which together mediate antigen specificity. However, high-throughput methods for analyzing the pairing of α and β chains have been elusive. Spindler et al. have removed this bottleneck by developing a droplet microfluidics-based system that maintains physical linkage between α and β TCR chains. TCR mRNA transcripts from individual T cells undergo reverse transcription into cDNA and subsequent amplification. Short linker DNA constructs attached to the amplification primers fuse the two TCR chains to ensure native pairing. αβ cDNA TCR libraries are then subcloned into lentiviral constructs for expression in Jurkat cells. This approach could elicit up to 100-fold more paired TCRs than previously described single-cell methods. The researchers generated multiple Jurkat libraries from donor peripheral blood samples, which allowed them to identify rare high-avidity antigen-specific TCR clonotypes in the context of cancer and viral infections. This method of high-resolution, high-throughput TCR repertoire analysis enhances our understanding of the dynamics of human αβ TCR repertoires and may facilitate the rapid discovery of rare TCRs for the development of efficient adoptive cell therapies.
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Mukhopadhyay, M. Paired TCR discovery in high throughput. Nat Methods 17, 462 (2020). https://doi.org/10.1038/s41592-020-0845-5