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Transcriptomics profiling of CD141+ dendritic cells isolated from peripheral blood or synovial fluid of arthritis patients

ABSTRACT: Transcriptomics profiling of CD141+ dendritic cells isolated from peripheral blood or synovial fluid of arthritis patients

PROVIDER: PRJNA422753 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6384380_1.fastq.gz	Fastqsanger.gz
	SRR6384380_2.fastq.gz	Fastqsanger.gz
	SRR6384390_1.fastq.gz	Fastqsanger.gz
	SRR6384390_2.fastq.gz	Fastqsanger.gz
	SRR6384381_1.fastq.gz	Fastqsanger.gz

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Transcriptomics profiling of CD141+ dendritic cells isolated from peripheral blood or synovial fluid of arthritis patients

Project description:Total RNA sequencing was performed on CD141+ DC isloated from peripheral blood of healthy individuals (n=5) or synovial fluid of patients with inflammatory arthritis (n=6)

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Transcriptomics profiling of CD141+ dendritic cells isolated from peripheral blood or synovial fluid of arthritis patients

Project description:Total RNA sequencing was performed on CD141+ DC isolated from peripheral blood of healthy individuals (n=4), synovial fluid of patients with inflammatory arthritis (n=5) and peripheral blood of patients with inflammatory arthritis (n=7). Cd1c+ DC were also sequenced from peripheral blood of healthy individuals (n=3).

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Project description:Rheumatoid arthritis (RA) is a chronic autoinflammatory disorder that affects small joints. Despite intense efforts, there isno definitive marker yet for early diagnosis RA and for monitoring the progression of this disease. We sought to catalog the proteins present in the synovial fluid of patients with rheumatoid arthritis. To identify lower abundance proteins, we undertook two approaches – we depleted the abundant proteins using a multiple affinity removal system (MARS14) column and we enriched glycoproteins using a lectin affinity column. The peptides were analyzed by LC-MS/MS on a high resolution Fourier transform mass spectrometer.

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Project description:Previously, extracellular vesicles (EVs) from rheumatoid arthritis (RA) synovial fluid (SF) have been reported to stimulate the release of pro-inflammatory mediators in recipient cells. However, mechanistic details are unclear and contaminants in EV enrichments might have compromised previous observations. We recently developed a novel, size exclusion chromatography (SEC)-based method of EV isolation capable of high-quality enrichments from human synovial fluid. Here, we employed this method in combination with high resolution mass spectrometry to accurately characterise the SF EV proteome and investigate contributions of SF EVs to inflammatory processes in RA.

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10x single cell gene expression and V(D)J immune profiling of T cells from the blood, synovial fluid and synovial tissue in psoriatic arthritis patients

Project description:This study shows the pronounced expansion of CD8 T cell clones at sites of active inflammation within the joints of psoriatic arthritis (PsA) patients compared to blood, and characterises the gene expression of expanded clones using droplet based single cell RNA sequencing. Mononuclear cells were separated from freshly acquired paired peripheral blood and synovial fluid samples from 3 PsA patients using density gradient separation, then enriched for CD4 and CD8 T cells using FACS. Additionally, CD45+ leukocytes were enriched by FACS from the cryopreserved synovial tissue of 2 further PsA patients. All cells were then processed using a 10x Chromium Controller and sequenced by Illumina HiSeq 4000 (peripheral blood / synovial fluid) or NovaSeq with S2 flowcell (synovial tissue).

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Project description:Psoriatic arthritis is a seronegative polyarticular form of inflammatory arthritis . Genetic analysis implicates a role for both IL-17/23 axis and CD8+ T cells in disease susceptibility. Using RNA-seq we identified differential gene expression between synovial IL-17A+(IFNy+/-) CD8+ T cells compared to IL-17A-IFNy+ CD8+ T cells and IL-17A+CD4+ T cells from the synovial fluid of psoriatic arthritis patients. We find that IL-17A+CD8+ T cells have a transcriptional overlap with IL-17A+CD4+ T cells. Overall we show these IL-17A+ CD8+ T cells have a polyfunctional, pro-inflammatory capacity and are potentially derived from common precursors, shared with IL-17A-CD8+ T cells.

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Project description:Objective. To analyse the pathogenesis of the diseases rheumatoid arthritis (RA) and spondyloarthritis (SpA) we investigated the protein composition in synovial fluid from the patients. Methods. Fifty-six synovial fluid (SF) were analysed with non-gel based proteomics from patients with RA (32) and SpA (24). Rheumatoid factor was measured in plasma, and cell-free DNA was measured in SF. Results. Two hundred proteins were quantified in the samples. The inflammatory proteins were more abundant in the RA group, specially proteins from neutrophil granulocytes. Cell-free DNA (cfDNA) in the SF was statistically associated with proteins that are known to form neutrophil extracellular traps (NETs). Plasma C-reactive protein (CRP), was correlated to other acute phase proteins in the SF. Minimal correlation was seen between acute phase proteins and proteins in NETs. Conclusions. The results show that in the synovial cavity in vivo neutrophils form NETs. This result sustains that neutrophils from RA patients are activated and are likely to undergo NETosis resulting in SF cfDNA specific of arthritis pathology with NETosis.

2021-09-08 | PXD010723 | Pride

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