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Mus musculus

ABSTRACT: Effect of immunization co-stimuli on transcriptome of mouse splenic IgG1+ germinal center B cells

PROVIDER: PRJNA352913 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5010532.fastq.gz	Fastqsanger.gz
	SRR5010533.fastq.gz	Fastqsanger.gz
	SRR5010534.fastq.gz	Fastqsanger.gz
	SRR5010535.fastq.gz	Fastqsanger.gz
	SRR5010536.fastq.gz	Fastqsanger.gz

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Project description:Effect of immunization co-stimuli on transcriptome of mouse splenic IgM− B220lo CD138+ plasma cells

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Expression data from murine eGFP+ relfl/flCg1-Cre and eGFP-Cg1-Cre splenic germinal center B cells purified by fluorescent activated cell sorting.

Project description:Gene expression profiling of murine eGFP+ relfl/flCg1-Cre and eGFP− Cg1-Cre splenic germinal center B cells identifies genes regulated by the transcription factor c-REL in germinal center B cells. Germinal center B cells from 12-week old relfl/flCg1-Cre and Cg1-Cre littermate mice immunized with sheep red blood cells (SRBC) were isolated at day 7 after immunization by flow cytometric sorting from splenic mononuclear cells. RNA was isolated. Following a cDNA amplification step, labeled cRNA was hybridized to microarrays and genes specifically expressed in eGFP+ relfl/flCg1-Cre and eGFP− Cg1-Cre samples were identified by supervised analysis.

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RNA-seq data from murine eGFP+ relbfl/flnfkb2fl/flCg1-Cre and Cg1-Cre splenic germinal center B cells purified by fluorescent activated cell sorting.

Project description:RNA-seq analysis of murine eGFP+ relbfl/flnfkb2fl/flCg1-Cre and Cg1-Cre splenic germinal center B cells identifies genes regulated by the transcription factors RELB and p52 (NF-kB2) in germinal center B cells. Germinal center B cells from 12-week old relbfl/flnfkb2fl/flCg1-Cre and Cg1-Cre littermate mice immunized with sheep red blood cells (SRBC) were isolated at day 7 after immunization by flow cytometric sorting from splenic mononuclear cells. RNA was isolated, amplified and submitted for RNA-sequencing on an Illumina HiSeq2500 instrument for 35-40 million 2x50 paired-ended reads.

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Transcriptome analysis of mouse IgG1 memory B cell subsets

Project description:IgE plays an essential role in the pathogenesis of allergies and its production is strongly regulated. A transient IgE germinal center phase and lack of IgE memory cells limit the generation of pathogenic IgE, but this can be overcome by sequential switching of IgG1 cells to IgE. We investigated which population of IgG1 cells can give rise to IgE-producing cells in memory responses. We identified three populations of IgG1 memory B cells (DP:CD73+CD80+, SP:CD73-CD80+, DN:CD73-CD80-) that generate IgE plasma cells of high or low affinity, but none gives rise to IgE germinal center cells or IgE memory cells. The two memory IgG1 populations differ however in their ability to differentiate into IgG1 plasma cells and germinal center cells, and to expand the IgG1 memory B cell pool. To explore the molecular mechanisms that may explain the distinct functions of IgG1 memory B cell subsets we compared their expression by transcriptome analysis using next generation sequencing.

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IL-21 Shapes the B Cell Response in a Context-Dependent Manner [RNA-seq

Project description:The cytokine interleukin-21 (IL-21) is a pivotal T cell-derived signal crucial for germinal center (GC) responses, but the precise mechanisms by which IL-21 influences B cell function remain elusive. Here, we investigated the B cell-intrinsic role of IL-21 signaling by employing a novel IL-21 receptor (Il21r) conditional knock-out mouse model and ex vivo culture systems and uncovered a surprising duality of IL-21 signaling in B cells. While IL-21 stimulation of naïve B cells led to Bim-dependent apoptosis, it promoted robust proliferation of pre-activated B cells, particularly class-switched IgG1+ B cells ex vivo. Consistent with this, B cell-specific deletion of Il21r led to a severe defect in IgG1 responses in vivo following immunization. Intriguingly, Il21r-deleted B cells are significantly impaired in their ability to transition from a pre-GC to a GC state following immunization. Although Il21r-deficiency did not affect the proportion of IgG1+ B cells among GC B cells, it greatly diminished the proportion of IgG1+ B cells among the plasmablast/plasma cell population. Collectively, our data suggest that IL-21 serves as a critical regulator of B cell fates, influencing B cell apoptosis and proliferation in a context-dependent manner.

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IL-21 Shapes the B Cell Response in a Context-Dependent Manner [CUT&RUN]

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Gene expression in germinal center light zone, dark zone, and naïve B cells

Project description:Microarrays of gene expression in mouse germinal center B cells photoactivated in the light zone or dark zone, and of naïve cells for comparison. We used microarray data to identify genes differentially expressed by B cells in the light and dark zones of the germinal center. Light and dark zone cells were photoactivated in day 7 germinal centers and sorted by flow cytometry. Naïve splenic B cells(IgD+CD19+) were sorted for comparison.

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