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Gene expression of resting memory, resting naive and in vitro activated memory CD8+CD127+ T cells from spleen and bone marrow (BM)

ABSTRACT: To understand differences between resting and activated memory CD8+ T cells, we compared the global gene expression of ex vivo isolated naive and spleen and BM memory cells to in vitro activated spleen and BM memory cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE62691 | GEO | 2015/02/05

SECONDARY ACCESSION(S): PRJNA264812

REPOSITORIES: GEO

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Gene expression of resting memory, resting naive and in vitro activated memory CD8+CD127+ T cells from spleen and bone marrow (BM)

Project description:To understand differences between resting and activated memory CD8+ T cells, we compared the global gene expression of ex vivo isolated naive and spleen and BM memory cells to in vitro activated spleen and BM memory cells. Single cell suspension from the spleen and bones of aged C57BL/6 mice were prepared. Naive (CD44-CD127+) and memory (CD44+CD127+) CD8+CD3+ T cells were then cytometrically sorted. Sorted cells were either immediately processed for RNA preparation or were activated with anti-CD3 and anti-CD28 for 42-44 hours. Total RNA was extracted using the NucleoSpin RNA (Macherey-Nagel). The integrity and amount of isolated RNA was assessed for each sample using an Agilent 2100 Bioanalyzer (Agilent, Waldbronn, Germany) and a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE). Double-stranded complementary RNA was synthesized from 1 M-BM-5g total RNA using Message AmpII Biotin (Ambion, USA). Fifteen micrograms of fragmented cRNA of each sample were hybridized to MG_U430_2 GeneChips (Affymetrix) in triplicates. Hybridization was performed in a Hybridization Oven 640, and chips were washed and stained in the Fluidics Station 400 (both Affymetrix). Finally, the arrays were scanned with a GeneChip Scanner 3000 using the GCOS software, version 1.4, both Affymetrix. All relevant GCOS data of quality checked microarrays were analyzed with High Performance Chip Data Analysis (HPCDA, unpublished), using the BioRetis database (www.bioretis-analysis.de), as described and validated previously.

2015-02-05 | E-GEOD-62691 | biostudies-arrayexpress

Whole-transcriptome profiling of LCMV.GP66-77 specific CD4 T cells isolated from bone marrow (BM) and spleen, 60 days after last immunization [RNA-seq]

Project description:To udnderstand the tissue-resident features of antigen-specific memory T cells of the bone marrow and spleen, we performed RNA-Seq and compared expression levels of genes of resting LCMV.GP66-77 specific CD4 T cells isolated from bone marrow (BM) and spleen of LCMV.GP61-80 primed C57BL/6 mice.

2019-02-11 | GSE124794 | GEO

Mus musculus

Project description:Gene expression of resting memory, resting naive and in vitro activated memory CD8+CD127+ T cells from spleen and bone marrow (BM)

| PRJNA264812 | ENA

Trancriptional comparison of Zbtb32-deficient and -sufficient polyclonal resting memory B cells

Project description:Memory B cell responses are more rapid and of greater magnitude than are primary antibody responses. The mechanisms by which these secondary responses are eventually attenuated remain unknown. We demonstrate that the transcription factor ZBTB32 limits the rapidity and duration of antibody recall responses. ZBTB32 is highly expressed by mouse and human memory B cells, but not by their naÃ¯ve counterparts. Zbtb32-/- mice mount normal primary antibody responses to T-dependent antigens. However, Zbtb32-/- memory B cell-mediated recall responses occur more rapidly and persist longer than do control responses. Microarray analyses demonstrate that Zbtb32-/- secondary bone marrow plasma cells display elevated expression of genes that promote cell cycle progression and mitochondrial function relative to wild-type controls. BrdU labeling and adoptive transfer experiments confirm more rapid production and a cell-intrinsic survival advantage of Zbtb32-/- secondary plasma cells relative to wild-type counterparts. ZBTB32 is therefore a novel negative regulator of antibody recall responses. Wild type and Zbtb32-/- resting polyclonal splenic memory B cells were purified by fluorescence activated cell sorting, RNA was extracted, and used for Affymetrix microarray analysis. 5 biological replicates of wild type and Zbtb32-/- cells were included for each cell type.

2016-06-10 | E-GEOD-83191 | biostudies-arrayexpress

Human memory T cells of the bone marrow are resting and maintain long-lasting memory

Project description:To compare human memory CD4+ T cell subsets in peripheral blood (PB) and bone marrow (BM) of healthy individuals at transcriptional level, we analyzed the global gene expression of ex vivo PB CD69- as well as BM CD69- and CD69+ memory CD4+ T cells from 4 paired PB and BM samples. The gene expression of these subsets was additionally compared to the transcriptional profile of 8 PB samples taken ex vivo or stimulated with phorbol myristate acetate (PMA) and Ionomycin for 3 hours.

2014-06-17 | GSE50677 | GEO

Molecular reprogramming of primary human resting memory CD4+ T cells by HIV-1 Vpr

Project description:Resting CD4+ T cells are infected by HIV-1 in vivo, however are refractory to cell-free HIV-1 infection in vitro. We show that they are efficiently infected by cell-to-cell spread. To allow resting T cell infection, uninfected resting target cells were co-cultured with infected donor T cells. Cells were infected with HIV-1 WT, dVpr or left mock treated and cultures with or without IL7. After 72h of co-culture, resting target cells were recovered by flow cytometry sorting and total RNA was extracted. RNASequencing revealed global transcriptomic reprogramming of resting memory T cells by HIV-1 Vpr.

2022-03-31 | E-MTAB-11454 | biostudies-arrayexpress

Gene profile beween CD19 normal and deletion of resting B cells in spleen of sirt1 conditional knockout mouse

Project description:Gene profile beween CD19 normal and deletion of resting B cells in spleen of sirt1 conditional knockout mouse

2019-05-01 | GSE121306 | GEO

Redirection of therapeutic T cells to a bone marrow niche improves their persistence and anti-tumor functions: transcription profiling by array of resting memory OT-I T_CXCR4) or OT-I T_Control cells isolated from the spleen 90 days following DC vaccination.

Project description:This study reports the improvement in function and antitumor efficacy of CXCR4 overexpression in therapeutic CD8+ T cells. Polyclonal CD8+ T cells from OT-I C57BL/6 (B6) mice were transduced with a modified pMP71 retroviral vector containing murine Cxcr4 and Gfp reporter sequences (T_CXCR4) or with a control vector containing Gfp alone (T_Control), and co-transferred into to B6 mice that were then vaccinated with peptide-pulsed DC. 90 days following DC vaccination, lymphocytes were isolated from the spleen and resting memory OT-I T_CXCR4 and OT-I T_Control cells were FACS sorted to perform gene expression profiling.

2018-03-28 | E-MTAB-6091 | biostudies-arrayexpress

Transcription profiling of mouse D44highCD62L-CD25- CD4+ T cells of murine (C57BL/6 mice) bone marrow were compared to those of the spleen

Project description:CD4+ T lymphocytes are key to immunological memory, but little is known about the lifestyle of memory CD4+ T lymphocytes. We showed that in the memory phase of specific immune responses to antigens, most of the memory CD4+ T lymphocytes relocated into the bone marrow (BM) within 3-8 weeks after their generation, a process involving integrin a2. Antigen-specific memory CD4+ T lymphocytes expressed Ly-6C to a high degree, unlike most splenic CD44hiCD62L- CD4+ T lymphocytes. In adult mice, more than 80% of Ly-6Chi CD44hiCD62L- memory CD4+ T lymphocytes were in the BM. In the BM, they are located next to IL-7-expressing VCAM-1+ stroma cells, and were in a resting state. Upon challenge with antigen, they rapidly expressed cytokines and CD154 and induced the production of high-affinity antibodies, indicating their functional activity in vivo and marking them as professional memory T helper cells Experiment Overall Design: FACSAria sorted CD44highCD62L-CD25- CD4+ T cells of murine (C57BL/6 mice) bone marrow were compared to those of the spleen using Affymetrix GeneChip Mouse Genome 430A 2.0 Array. After total RNA extraction, reverse transcription, cDNA extraction, the biotinylated cRNA was transcribed, fragmented, and 15 Âµg cRNA hybridized in triplicates for each of the two groups to the GeneChip arrays. Group of bone marrow chips: BMCD4T1, BMCD4T2, BMCD4T3, group of spleen chips: SCD4T1, SCD4T2, SCD4T3. Lists of differentially regulated genes were created using High Performance Chip Data Analysis (HPCDA) with Bioretis database (http://www.bioretis-analysis.de).

2009-04-25 | E-GEOD-15733 | biostudies-arrayexpress

Single cell RNA-seq analysis for early erythroblasts in spleen and bone marrow (BM) with or without mitochondria transfer

Project description:Single cell RNA-seq analysis for early erythroblasts in spleen and BM with or without mito-Dendra2 fluorescence signal after macrophage infusion upon PHZ treatment

2022-09-08 | GSE185267 | GEO

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