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TCR sequencing paired with massively parallel 3′ RNA-seq reveals clonotypic T cell signatures

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High-throughput 3′ single-cell RNA-sequencing (scRNA-seq) allows cost-effective, detailed characterization of individual immune cells from tissues. Current techniques, however, are limited in their ability to elucidate essential immune cell features, including… Click to show full abstract

High-throughput 3′ single-cell RNA-sequencing (scRNA-seq) allows cost-effective, detailed characterization of individual immune cells from tissues. Current techniques, however, are limited in their ability to elucidate essential immune cell features, including variable sequences of T cell antigen receptors (TCRs) that confer antigen specificity. Here, we present a strategy that enables simultaneous analysis of TCR sequences and corresponding full transcriptomes from 3′-barcoded scRNA-seq samples. This approach is compatible with common 3′ scRNA-seq methods, and adaptable to processed samples post hoc. We applied the technique to identify transcriptional signatures associated with T cells sharing common TCRs from immunized mice and from patients with food allergy. We observed preferential phenotypes among subsets of expanded clonotypes, including type 2 helper CD4+ T cell (TH2) states associated with food allergy. These results demonstrate the utility of our method when studying diseases in which clonotype-driven responses are critical to understanding the underlying biology. Current high-throughput 3′ single-cell RNA-sequencing (scRNA-seq) techniques are limited in their ability to elucidate the variable sequences of antigen receptors. Love and colleagues describe a strategy that enables simultaneous analysis of TCR sequences and the corresponding full transcriptomes from 3′-barcoded scRNA-seq samples.

Keywords: seq; scrna seq; sequencing paired; tcr sequencing; rna; cell

Journal Title: Nature Immunology
Year Published: 2019

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