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Single-cell RNA and TCR sequencing of sorted CD4+CD26- T cells sorted from 7 CTCL peripheral blood samples

ABSTRACT: single-cell RNA and TCR sequencing of sorted CD4+CD26- T cells sorted from 7 CTCL peripheral blood samples

PROVIDER: PRJNA856676 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20033060_1.fastq.gz	Fastqsanger.gz
	SRR20033060_2.fastq.gz	Fastqsanger.gz
	SRR20033061_1.fastq.gz	Fastqsanger.gz
	SRR20033061_2.fastq.gz	Fastqsanger.gz
	SRR20033062_1.fastq.gz	Fastqsanger.gz

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single-cell RNA and TCR sequencing of sorted CD4+CD26- T cells sorted from 7 CTCL peripheral blood samples

Project description:Purpose: Characterization of the gene expression profile and TCR profile of CD4+CD26- T cells from CTCL to understand the cell of origin

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Project description:Transcriptomic profiling of ovine skin lymph dendritic cell subsets CD26+ and CD26- compared to total enriched lymph dendritic cells. CD26+ and CD26- dendritic cells (DC) from sheep skin lymph were sorted by flow cytometry and were analysed for their transcriptomic profile. We demonstrate that the minor sheep CD26+ skin lymph DC subset shares significant transcriptomic similarities with mouse CD8a+ and human BDCA3+ DC. This allowed the identification of a common set of phenotypic characteristics for CD8a-like DC in the 3 mammalian species, i.e. SIRPlo, CADM1hi, CLEC9Ahi, DEC205hi, XCR1hi. Compared to CD26- DC, the sheep CD26+ DC show (1) potent stimulation of allogeneic naive CD8+ T cells with high selective induction of the Ifn-gamma and Il22 genes; (2) dominant efficacy in activating specific CD8+ T cells against exogenous soluble antigen; (3) selective expression of functional pathways associated to high capacity for antigen cross-presentation. Reference design (total enriched lymph dendritic cells) - Biological replicates: 2 sheep #10081 and #30066

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Project description:Sezary syndrome (SS) represents a leukemic variant of cutaneous T cell lymphoma (CTCL) with circulating malignant CD4 T cells trafficking to the skin. The cell surface molecules present on malignant cells are also expressed on normal CD4 T cells. Therefore, we attempted to find a specific marker for malignant cells that would distinguish them from normal cells. Comprehensive microarray analysis of gene expression in the malignant cells indicated significantly increased levels of mRNA for cell surface markers CD164, a sialomucin found on human CD34+ hematopoietic stem cells, as well as FCRL3, a molecule present on a subset of human natural T regulatory cells. Both markers were increased in CD4 T cells from the SS patients compared to those from Mycosis Fungoides patients and healthy volunteers. Subsequent studies utilizing QPCR and flow cytometry confirmed the increased expression of CD164 and FCRL3 mainly in CD4+CD26- T cells of SS patients. Our results suggest that CD164 can serve as a marker for diagnosis as well as disease monitoring of CTCL/SS, whereas the FCRL3 expression correlates with disease progression.

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