Unknown,Transcriptomics,Genomics,Proteomics

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Furin deficiency efect on T cells

ABSTRACT: Analysis of T-cells lacking the proprotein convertase furin. Proprotein convertases promote the proteolytic maturation of proproteins. Furin is induced in activated T-cells. Results provide insight into the function of furin in T-cells. CD4+CD62L+CD44- naive, CD4+CD62L-CD44+ memory and CD4+CD25+FoxP3+ regulatory T cells were isolated from Fur flox/flox and CD4 cre Fur flox/flox mice. Naive T cells were activated via TCR. Total RNA was extracted from all cells and hybridized to Affymetrix microarrays.

ORGANISM(S): Mus musculus

SUBMITTER: Marko Pesu

PROVIDER: E-GEOD-67884 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Proprotein convertase FURIN regulates T cell receptor-induced transactivation.

Ortutay Zsuzsanna Z Oksanen Anna A Aittomäki Saara S Ortutay Csaba C Pesu Marko M

Journal of leukocyte biology 20150429 1

Antigen emergence rapidly stimulates T cells, which leads to changes in cytokine production, cell proliferation, and differentiation. Some of the key molecules involved in these events, such as TGF-β1 and NOTCH1, are synthesized initially as inactive precursors and are proteolytically activated during T cell activation. PCSKs regulate proprotein maturation by catalyzing the proteolytic cleavage of their substrates. The prototype PCSK FURIN is induced upon TCR activation, and its expression in T ...[more]

PMID: 25926688

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Project description:Purpose: FURIN is a member of the proprotein convertase subtilisin/kexin (PCSK) family of serine endoproteases important in converting immature proproteins into their functional form. While FURIN is important in various aspects of the immune response such as in CD4+ T cells, FURIN´s role in CD8+ T cells is unclear. Here, we isolated naive CD8+ T cells from conditional T cell specific Furin KO (Cd4-Cre-Furinflox/flox) and WT (Furinflox/flox) mice and studied the genome-wide gene expression pattern upon in vitro anti-CD3/anti-CD28 activation (plate-bound, 5µg/ml each) using RNA sequencing. We also used recombinant TGFB1 (0.5ng/ml) or IL12 (10ng/ml) in the same experimental setting to study their impact on the CD8+ T cell transcriptome. Methods: Total RNA was isolated from the stimulated CD8+ T cells and the RNA-sequencing data was produced by the Functional Genomics Unit of the HiLIFE Genome Analysis Infrastructure (University of Helsinki, Helsinki, Finland) using Illumina NextSeq High Output 1x75 bp. Data processing pipeline was built on Snakemake wrappers, and used STAR to quantify features from quality and adapter trimmed reads in gene level. Prior the normalisation and differential gene expression analysis with DESeq2, the matrix of raw gene counts was prefiltered to keep only rows that have at least 10 reads in total. Result tables were annotated with biomaRt. Results: Snakemake preprocessing pipeline measured 55573 genes against GRCm38, ENSEMBL release 97. 17714 of them was used for statistical testing. Full result tables for each treatment were ranked by p-value and the normalized DESeq2 abundances in samples were included. The genes with BH-adjusted p-value <0.05 was considered differentially expressed. Conclusions: Our study revealed that the abscence of FURIN influences the expression of several genes in activated CD8+ T cells. Additionally, although FURIN-deficient CD8+ T cells can respond to exogenous TGFB1 and IL12, administrating these cytokines further increases the number of differentially expressed genes between Furin KO and WT CD8+ T cells.

Dataset Information

Furin deficiency efect on T cells

Publications

Proprotein convertase FURIN regulates T cell receptor-induced transactivation.

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