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Transcriptomic analysis of naïve B cells and antigen-specific activated B cells and plasma cells from 4dpi NP-LPS immunized mice [NPLPS_PC]

ABSTRACT: Little is known about the earliest transcriptomic events in plasma cell differentiation. Indeed, activation of UPR-affiliated gene expression is seen in in-vitro plasma cell differentiation prior to Blimp1 upregulation and in the absence of Xbp1 as we show in linked companion series. In this study, Blimp1 reporter mice were immunized with the T-independent antigen NP-LPS four days prior to spleen harvest and naïve B cells, NP-binding activated B cells and NP-binding plasma cells were analyzed by bulk RNA-seq. Here we report the activation of ER remodeling gene expression in antigen-specific activated B cells in-vivo prior to upregulation of Blimp1 transcription.

ORGANISM(S): Mus musculus

PROVIDER: GSE141420 | GEO | 2019/12/05

REPOSITORIES: GEO

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Transcriptomic analysis of in-vitro plasma cell generation from follicular B cells lacking Xbp1 [Xbp1_PC_stim]

Project description:During plasma cell differentiation there is activation of UPR gene expression that has been termed the physiological UPR of plasma cell differentiation. This is canonically thought to be downstream of Blimp1-mediated increases in immunoglobulin gene production and activation of the Xbp1 transcription factor by the RNA splicing activity of the ER-stress sensor IRE1α. Having observed UPR-affiliated gene expression prior to upregulation of Blimp1 as reported in the companion linked series, we endeavored to determine the necessity of Xbp1 activity in the early remodeling of the ER in nascent plasma cells. To this end, we mated mice harboring a floxed exon 2 of the Xbp1 gene (Xbp1flox) to mice expressing a tamoxifen-inducible cre recombinase under the control of the human CD20 promoter (hCD20-TamCre). These mice and their hCD20-TamCre-Xbp1wt litermates were fed oral tamoxifen in their diet for two weeks and follicular B cells were prepared for in-vitro plasma cell differentiation studies. We report here that prior to plasma cell differentiation as measured by CD138 expression, which coincides temporally with Blimp1 expression, there is no defect in UPR-affiliated gene expression in cells lacking Xbp1.

2019-12-05 | GSE141422 | GEO

Transcriptomic analysis of in-vitro plasma cell generation from follicular B cells lacking Raptor [Raptor_PC_stim]

Project description:Early UPR-affiliated gene expression occurs before Blimp1 upregulation and independent of canonical ER-stress activated Xbp1. In linked companion studies we detailed how Xbp1-independent activation of early plasma cell specific UPR-affiliated gene expression occurs prior to Blimp1 upregulation. Having observed that these genes overlapped with canonical mTORC1 signature genes and that PC-poised marginal zone B cells have higher base line mTORC1 signaling we designed this experiment to assess the dependence of early ER-remodeling on mTORC1 signaling in plasma cell differentiation. To this end we mated mice harboring a floxed allele for the mTORC1 adapter Raptor (B6.Rptor-flox) to mice expressing a tamoxifen-inducible cre recombinase under the control of the human CD20 promoter (B6.hCD20-TamCre). These mice and their hCD20-TamCre-Rptor-wt litermates were fed oral tamoxifen in their diet for two weeks and follicular B cells were prepared for in-vitro plasma cell differentiation studies. We report here that as early as 24 hours of stimulation in culture we see a marked defect in the ability of Rptor null B cells to upregulate UPR-affiliated genes associated with plasma cell differentiation.

2019-12-05 | GSE141421 | GEO

Transcriptomic analysis of naïve B cells and plasma cells as well as in vitro stimulated B cells from Blimp1-GFP reporter mice [Blimp1_reporter_PC_stim]

Project description:Though it is known that plasma cells utilize the unfolded protein response to remodel their ER it is generally believed that this follows Blimp1-mediated transcriptional and translational increases in immunoglobulin protein. In this study we utilize Blimp1 reporter mice to examine the earliest activation of UPR-affiliated gene expression in plasma cell inductive stimulation of B cells. We combine in-vivo gene expression of follicular B cells, marginal zone B cells from spleen and short and long lived plasma cells from bone marrow with in-vivo generated activated B cells and plasma cells from the same mice to examine gene expression before and after Blimp1-upregulation. Using a novel in-vitro stimulation approach we separate those UPR-affiliated genes activated by B cell activation and those requiring a plasma cell inductive stimulus. We report here that these plasma cell specific UPR-affiliated genes are activated in absence of ER-stress prior to upregulation of Blimp1.

2019-12-05 | GSE141419 | GEO

Next generation sequencing of antibody heavy chain genes that bind to the hapten NP in sorted memory cells and long-lived plasma cells isolated from mice immunized orally with NP-CT

Project description:We report that there are differences in V gene usage in memory B cells and long-lived plasma cells after oral immunization. The hapten NP was conjugated to cholera toxin (CT) to create NP-CT, an antigen that is highly immunogenic after oral immunization. The NP hapten induces immune responses dominated by the 1-72 (VH186.2) heavy chain V region. The relationship between NP binding IgA antibody genes from memory B cells from spleen, MLN and Peyer´s patches and long-lived plasma cells from lamina propria and bone marrow that persisted 6-12 months after an oral immunization in three C57BL/6 mice with NP-CT was investigated. Extensive clonal overlap im clones was observed between long-lived memory cells from lamina propria and bone marrow, but very limited overlap was found between memory cells and plasma cells. The data suggest that memory and plasma cells formed through temporarily or anatomically separate processes.

2016-08-18 | GSE84698 | GEO

Project description:NP Metagenome

| PRJNA724020 | ENA

Transcription profiling of mouse naive B cells vs. antigen-specific plasma, germinal center, and memory B cells after immunization with the T-dependent antigen, NP-CGG, in order to generate a transcriptional profile of self-renewal within the hematopoietic system

Project description:In order to better understand the factors that regulate B cell differentiation upon exposure to antigen, we compares global gene expression profiles from naive B cells with antigen-specific plasma, germinal center, and memory B cells after immunization with the T-dependent antigen, NP-CGG. The memory B cell-enriched transcripts were then compared with memory T cell-enriched and hematopoietic stem cell-enriched transcripts in order to generate a transcriptional profile of self-renewal within the hematopoietic system.

2007-12-15 | E-GEOD-4142 | biostudies-arrayexpress

NP-18 vs. NP-18AR transcriptional profile upon treatment with TK/GCV

Project description:Pancreatic cancer-derived cells NP-18 underwent four rounds of treatment with increasing doses of an adenoviral vector encoding TK enzyme and GCV. Surviving cells were termed NP-18AR and displayed decreased sensitivity to treatment. This experiment analyses the transcriptomic effect of TK/GCV treatment on NP-18AR as compared to that of NP-18 cells. Two-condition experiment, where response to treatment in naïve cells (NP-18) was compared to response to treatment in previously treated resistant cells (NP-18AR). Biological replicates: samples from 3 independently generated and independently treated NP-18AR lines (NP-18AR1, NP-18AR2 and NP-18AR3) were hibridized with 3 independently treated samples of NP-18 cells. A total of 9 hybridizations were performed, 2 for NP-18AR1 (including 1 dye-swap), 4 for NP-18AR2 (including 2 dye swaps) and 3 for NP-18AR3 (including 1 dye-swap).

2010-07-16 | E-GEOD-17137 | biostudies-arrayexpress

NP-18 vs. NP-18AR transcriptional profile upon treatment with TK/GCV

2010-07-16 | GSE17137 | GEO

Comparative global gene expression profiling of mice liver and spleen fed with nucleoprotein (NP) diets

2011-04-13 | GSE28505 | GEO

Comparative global gene expression profiling of mice liver and spleen fed with nucleoprotein (NP) diets

Project description:A diet rich in nucleic acids and protamin protein, termed as nucleoprotein was used for the study. Mice were fed with NP diets for 4 weeks followed by removal of the liver and spleen. Total RNA extracted from livers and spleens was pooled in each group (low NP or LNP-control, and 1.2% NP-treatment, diets), prior to DNA microarray analysis (Agilent mouse whole genome 4 x 44K). Results revealed 1373 & 3386 up (>1.5 fold)- and down (<0.75 fold)-regulated genes in the liver, and 252 & 1838 up- and down-regulated genes in the spleen, respectively following 1.2%NP diet. Analysis of genes related to NP diets will be discussed. To investigate the effect of NP, we added S-nuclegen® at a concentration of 1.2% into CLEA basic purified diet (CLEA JAPAN, Inc., Tokyo, Japan) known to include nucleic acids (NAs) at much lower amounts than standard diet. By HPLC analysis CLEA basic purified diet (low NP), and 1.2% NP diet included 0.03%, and 0.5% NAs, respectively. Male C57BL/6J mice (13 weeks) were fed with NP diet for 4 weeks. The liver and spleen were removed and deep frozen in liquid nitrogen. Whole blood was also sampled from these mice, and frozen in liquid nitrogen. Total RNA extracted from livers and spleens was pooled in each group (control, lowNP or LNP; and treatment, 1.2%NP), prior to DNA microarray analysis (Agilent mouse whole genome 4 x 44K).

2011-04-13 | E-GEOD-28505 | biostudies-arrayexpress

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