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Canine peripheral blood TCRαβ T cell atlas: Identification of diverse subsets including CD8+ MAIT-like cells by combined single-cell transcriptome and V(D)J repertoire analysis

ABSTRACT: Canine peripheral blood TCRαβ T cell atlas: Identification of diverse subsets including CD8+ MAIT-like cells by combined single-cell transcriptome and V(D)J repertoire analysis

PROVIDER: PRJNA903202 | ENA |

REPOSITORIES: ENA

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Canine peripheral blood TCRαβ T cell atlas: Identification of diverse subsets including CD8+ MAIT-like cells by combined single-cell transcriptome and V(D)J repertoire analysis

Project description:The diversity of canine T cells is poorly understood. We used 5’ scRNA-seq coupled with immune repertoire sequencing to generate a highly detailed atlas of peripheral blood TCRαβ+ T cells of healthy dogs.

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Canine peripheral blood TCRαβ T cell atlas: Identification of diverse subsets including CD8A<sup>+</sup> MAIT-like cells by combined single-cell transcriptome and V(D)J repertoire analysis.

Project description:The dog is valued as a companion animal and increasingly recognized as a model for human disorders. Given the importance of T cells in health and disease, comprehensive knowledge of canine T cells can contribute to our understanding of pathogenesis mechanisms and inform the development of new treatment strategies. However, the diversity of canine T cells is still poorly understood mainly due to the lack of species-reactive antibodies for use in flow cytometry. The aim of this study was to generate a detailed atlas of peripheral blood TCRαβ+ T cells of healthy dogs using single-cell RNA-sequencing (scRNAseq) combined with immune repertoire sequencing. A total of 22 TCRαβ+ T cell clusters were identified, which were classified into three major groups: CD4-dominant (11 clusters), CD8A-dominant (8 clusters), and CD4/CD8A-mixed (3 clusters). Based on differential gene expression, distinct differentiation states (naïve, effector, memory, exhausted) and lineages (e.g. CD4 T helper and regulatory T cells) could be distinguished. Importantly, several T cell populations were identified, which have not been described in dogs before. Of particular note, our data provide first evidence for the existence of canine mucosa-associated invariant T cell (MAIT)-like cells, representing one of three newly identified FCER1G+ innate-like CD8A+ T cell populations in the peripheral blood of healthy dogs. In conclusion, using scRNAseq combined with immune repertoire sequencing we were able to resolve canine TCRαβ+ T cell populations at unprecedented resolution. The peripheral blood TCRαβ+ T cell atlas of healthy dogs generated here represents an important reference data set for future studies and is of relevance for identifying new targets for T cell-specific therapies.

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Parallel T-cell cloning and deep sequencing of the transcripts of human MAIT cells reveal stable oligoclonal TCRβ repertoire

Project description:Mucosal-associated invariant T (MAIT) cells are abundant in humans and recognize conserved bacterial antigens derived from riboflavin precursors, presented by the non-polymorphic MHC class I-like molecule MR1. Here, we show via transcriptomic analysis that human MAIT cells are remarkably oligoclonal in both blood and liver, display high inter-individual homology, and exhibit a restricted length CDR3β domain of the TCRVβ chain. We extend this analysis to a second sub-population of MAIT cells expressing a semi-invariant TCR conserved between individuals. Study of CDR3 regions of TCRalpha and beta sequences

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