期刊
BIOINFORMATICS
卷 38, 期 14, 页码 3645-3647出版社
OXFORD UNIV PRESS
DOI: 10.1093/bioinformatics/btac382
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资金
- Ludwig Center at Harvard
- NIH [R00CA218832]
- Gilead Sciences
- Bertarelli Rare Cancers Fund
- William Guy Forbeck Research Foundation
- Glenn Foundation for Medical Research
- American Federation for Aging Research (AFAR) Grant
- Government of Spain [FPU18/05488]
This study presents a computational pipeline to process TCR-enriched data from single-cell RNA sequencing. The tool enables the reconstruction of T-cell clonotypes and associated transcriptomes, and detects a high proportion of TCR sequences in single T cells from a healthy donor. Additionally, the study finds that TCR clones are larger in CD8 Memory T cells.
Diversity of the T-cell receptor (TCR) repertoire is central to adaptive immunity. The TCR is composed of alpha and beta chains, encoded by the TRA and TRB genes, of which the variable regions determine antigen specificity. To generate novel biological insights into the complex functioning of immune cells, combined capture of variable regions and single-cell transcriptomes provides a compelling approach. Recent developments enable the enrichment of TRA and TRB variable regions from widely used technologies for 3'-based single-cell RNA-sequencing (scRNA-seq). However, a comprehensive computational pipeline to process TCR-enriched data from 3' scRNA-seq is not available. Here, we present an analysis pipeline to process TCR variable regions enriched from 3' scRNA-seq cDNA. The tool reports TRA and TRB nucleotide and amino acid sequences linked to cell barcodes, enabling the reconstruction of T-cell clonotypes with associated transcriptomes. We demonstrate the software using peripheral blood mononuclear cells from a healthy donor and detect TCR sequences in a high proportion of single T cells. Detection of TCR sequences is low in non-T-cell populations, demonstrating specificity. Finally, we show that TCR clones are larger in CD8 Memory T cells than in other T-cell types, indicating an association between T-cell clonotypes and differentiation states.
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