TCR+/BCR+ dual-expressing cells and their associated public BCR clonotype are not enriched in type 1 diabetes

Japp, Alberto Sada; Meng, Wenzhao; Rosenfeld, Aaron M.; Perry, Daniel J.; Thirawatananond, Puchong; Bacher, Rhonda; Liu, Chengyang; Gardner, Jay; Atkinson, Mark A.; Kaestner, Klaus H.; Brusko, Todd M.; Naji, Ali; Prak, Eline T. Luning; Betts, Michael R.

doi:10.1016/j.cell.2020.11.035

Cited by 20 publications

(19 citation statements)

References 62 publications

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“…described CD5 expression as a marker of DE cells and detected higher proportions in T1D patients as compared with controls. These findings were recently challenged by Japp et al 16 . who did not find higher proportions of DE cells in T1D patients and questioned lineage integrity of DE cells.…”

Section: Discussionmentioning

confidence: 94%

CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls

et al. 2021

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Different lymphocyte subsets are involved in autoimmune pathogenesis of type 1 diabetes (T1D). Previous studies suggested a role of CD5‐expressing T and B cells including rare unconventional lymphocytes with combined T‐ and B‐cell features [dual expressing (DE) cells]. We performed algorithm‐supported multiparameter flow cytometry and quantitative PCR to investigate immune cell subsets and DE cells in children with T1D (n = 20) and matched controls (n = 20). Comparisons of conventional immune cells detected increased proportions of CD3+ T cells in T1D patients, whereas CD19+ B‐cell proportions were comparable to controls. Self‐organizing maps for flow cytometry analyses (FlowSOM) showed highly similar CD5‐expressing B‐cell subsets and no differences for DE cells were detected between the study groups by flow cytometry or specific quantitative PCR. Notably, differences in CD8+ T cells were indicated by FlowSOM and similarity‐based t‐distributed stochastic neighbor embedding (tSNE) analyses. Study group comparisons confirmed significantly reduced CD8+ T‐cell proportions with moderate or low CD5 expression in T1D patients. Finally, in vitro experiments showed stable CD5 expression differences of CD8+ T cells after T‐cell activation, cytokine stimulation and culture. We observed differences of T‐cell coreceptor CD5 expression in T1D patients with potential relevance for immune regulation of CD8+ T‐cell activation.

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Section: Discussionmentioning

confidence: 94%

CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls

et al. 2021

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“…Dual expression of Ig and TCRs or other B and T cell markers have been reported by others, but the role that these hybrid lymphocytes play is largely unknown. 33-35 The possibility of doublets, where two cells are given the same cellular barcode, does arise when using droplet-based single cell technologies such as the 10x Genomics used here. To assess the potential for these cells to be B+T cell doublets we used differential expression analysis to identify transcriptomic signatures that were absent in B and T cells.…”

Section: Resultsmentioning

confidence: 99%

Single cell based high-throughput Ig and TCR repertoire sequencing analysis in rhesus macaques

Tollison

Brochu

et al. 2021

Preprint

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Recent advancements in microfluidics and high-throughput sequencing technologies have enabled recovery of paired heavy- and light- chains of immunoglobulins (Ig) and VDJ- and VJ- chains of T cell receptors (TCR) from thousands of single cells simultaneously in humans and mice. Despite rhesus macaques being one of the most well-studied model organisms for the human adaptive immune response, high-throughput single cell immune repertoire sequencing assays are not yet available due to the complexity of these polyclonal receptors. Here we employed custom primers that capture all known rhesus macaque Ig and TCR isotypes and chains that are fully compatible with a commercial solution for single cell immune repertoire profiling. Using these rhesus specific assays, we sequenced Ig and TCR repertoires in over 60,000 cells from cryopreserved rhesus PBMC, splenocytes, and FACS-sorted B and T cells. We were able to recover every Ig isotype and TCR chain, measure clonal expansion in proliferating T cells, and pair Ig and TCR repertoires with gene expression profiles of the same single cells. Our results establish the ability to perform high-throughput immune repertoire analysis in rhesus macaques at the single cell level.

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“…It is suggested that these "dual expressors" (DE) are increased in frequency in type 1 diabetes and that in people with type 1 diabetes there is a public BCR which can stimulate insulin-reactive CD4+ T cells. However, this work remains controversial as others have been unable to replicate the enrichment of DE in T1D nor the specific public BCR sequence (75). This highlights the importance of good quality control at every step of scRNAseq experiments.…”

Section: Identifying Islet-reactive Bcrs In the Periphery Pancreatic Lymph Nodes And Pancreasmentioning

confidence: 99%

Insights From Single Cell RNA Sequencing Into the Immunology of Type 1 Diabetes- Cell Phenotypes and Antigen Specificity

et al. 2021

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In the past few years, huge advances have been made in techniques to analyse cells at an individual level using RNA sequencing, and many of these have precipitated exciting discoveries in the immunology of type 1 diabetes (T1D). This review will cover the first papers to use scRNAseq to characterise human lymphocyte phenotypes in T1D in the peripheral blood, pancreatic lymph nodes and islets. These have revealed specific genes such as IL-32 that are differentially expressed in islet –specific T cells in T1D. scRNAseq has also revealed wider gene expression patterns that are involved in T1D and can predict its development even predating autoantibody production. Single cell sequencing of TCRs has revealed V genes and CDR3 motifs that are commonly used to target islet autoantigens, although truly public TCRs remain elusive. Little is known about BCR repertoires in T1D, but scRNAseq approaches have revealed that insulin binding BCRs commonly use specific J genes, share motifs between donors and frequently demonstrate poly-reactivity. This review will also summarise new developments in scRNAseq technology, the insights they have given into other diseases and how they could be leveraged to advance research in the type 1 diabetes field to identify novel biomarkers and targets for immunotherapy.

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TCR+/BCR+ dual-expressing cells and their associated public BCR clonotype are not enriched in type 1 diabetes

Cited by 20 publications

References 62 publications

CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls

CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls

Single cell based high-throughput Ig and TCR repertoire sequencing analysis in rhesus macaques

Insights From Single Cell RNA Sequencing Into the Immunology of Type 1 Diabetes- Cell Phenotypes and Antigen Specificity

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TCR+/BCR+ dual-expressing cells and their associated public BCR clonotype are not enriched in type 1 diabetes

Cited by 20 publications

References 62 publications

CD5‐expressing CD8+ T‐cell subsets differ between children with type 1 diabetes and controls

CD5‐expressing CD8+ T‐cell subsets differ between children with type 1 diabetes and controls

Single cell based high-throughput Ig and TCR repertoire sequencing analysis in rhesus macaques

Insights From Single Cell RNA Sequencing Into the Immunology of Type 1 Diabetes- Cell Phenotypes and Antigen Specificity

Contact Info

Product

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CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls

CD5‐expressing CD8⁺ T‐cell subsets differ between children with type 1 diabetes and controls