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Transcription profile of B cells isolated from peripheral lymph nodes of wild-type vs CCR7-/- mice

ABSTRACT: We investigated the transcriptional profile of B cells isolated from peripheral lymph nodes of CCR7-/-C57BL/6 and C57BL/6 mice respectively. B cells were sorted as follows: CD19+, CD3-, CD11c-, CD11b-, NK1.1-, GR-1-. Very few genes were differentially regulated in CCR7-/- B cells, however the expression of a number of genes associated with B-cell activation was increased in CCR7-/- B cells compared to WT B cells. Our data suggest a role of CCR7 signaling in B-cell activation processes.

ORGANISM(S): Mus musculus

PROVIDER: GSE27885 | GEO | 2011/11/21

SECONDARY ACCESSION(S): PRJNA137997

REPOSITORIES: GEO

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Transcription profile of B cells isolated from peripheral lymph nodes of wild-type vs CCR7-/- mice

Project description:We investigated the transcriptional profile of B cells isolated from peripheral lymph nodes of CCR7-/-C57BL/6 and C57BL/6 mice respectively. B cells were sorted as follows: CD19+, CD3-, CD11c-, CD11b-, NK1.1-, GR-1-. Very few genes were differentially regulated in CCR7-/- B cells, however the expression of a number of genes associated with B-cell activation was increased in CCR7-/- B cells compared to WT B cells. Our data suggest a role of CCR7 signaling in B-cell activation processes. 12 purified RNA samples (6 samples of each genotype), originated from 12 individually facs-sorted cell samples, were used for the microarray study. 3 RNA samples per genotype were pooled, giving rise to 2 wildtype- and 2 Ccr7 knock-out pools. In each of the 2 dual-color microarray hybridizations, samples from 1 wildtype and 1 Ccr7 knock-out pool were cohybridized. Microarray hybridizations 1 and 2 were performed in a dye-swap approach.

2011-11-21 | E-GEOD-27885 | biostudies-arrayexpress

Single-cell RT-PCR analysis of Tgd17 progenitors

Project description:The indicated thymic progenitor population was sorted via FACS and then loaded into a Fluidigm C1 small cell capture chip for single-cell capture, lysis, reverse transcription, and preamplification. Preamplified products were analyzed on a BioMark HD with EvaGreen chemistry. The genes analyzed were selected based on bulk RNA sequencing data and/or prior publications, including genes relevant for gamma/delta T cells and T cell progenitors. B6=C57BL/6J, Rag=B6.Rag1-/-, Tcrd=B6.Tcrd-/-, DN1d=Live/TCRd-/Lin-/CD44+/CD25-/CD24+/cKit-, DN2=Live/TCRd-/Lin-/CD44+/CD25+/cKit+, cKit-=Live/TCRd-/Lin-/CD44+/CD25-/cKit-. Lin=CD3/CD8/CD11b/CD11c/CD19/Gr-1/NK1.1/TCRb/Ter-119 Sample naming nomenclature is as follows: MouseLine_CellPopoulation_DateCaptured_CaptureChamber_CellNumberInChamber

2018-08-21 | GSE118770 | GEO

Gene expression profiling of naïve bone marrow-resident granulocyte monocyte precursors (GMPs) and TSLP-elicited splenic GMP-like cells

Project description:Extramedullary hematopoiesis (EMH) refers to the differentiation of hematopoietic stem cells (HSCs) into effector cells that occurs in compartments outside of the bone marrow. Previous studies linked pattern recognition receptor (PRR)-expressing HSCs, EMH and immune responses to microbial stimuli. However, the factors that regulate EMH and whether EMH operates in broader immune contexts remain unknown. Here, we demonstrate a previously unrecognized role for thymic stromal lymphopoietin (TSLP) in promoting the population expansion of progenitor cells in the periphery and identify that TSLP-elicited progenitors differentiate into effector cells including macrophages, dendritic cells and granulocytes that contribute to TH2 cytokine responses. The frequency of circulating progenitor cells was also increased in allergic patients with a gain-of-function polymorphism in TSLP, suggesting the TSLP-EMH pathway may operate in human disease. These data identify that TSLP-induced EMH contributes to the development of allergic inflammation and indicate that EMH is a conserved mechanism of innate immunity. In this study, we performed gene expression profiling to examine how the transcriptional signatures compared between TSLP-elcited GMP-like cells and naïve bone marrow-resident GMPs. Splenic TSLP-elicited GMP-like cells (CD3-, CD4-, CD5-, CD8-, CD19-, CD11b-, CD11c-, NK1.1-, FcεRI-, CD34+ c-kit+) were sort-purified from the spleen of C57BL/6 mice on day 5 post-TSLP-cDNA treatment. BM-resident GMPs (CD3-, CD4-, CD5-, CD8-, CD19-, CD11b-, CD11c-, NK1.1-, FcεRI-, Sca1- c-kit+ CD34+ CD16/32+) were sort-purified from mice that received a control-cDNA injection.

2014-03-28 | E-GEOD-52485 | biostudies-arrayexpress

Gene expression profiling of Bone Marrow FoxP3+ Treg cells

Project description:Gene expression profiling of Bone Marrow FoxP3+ Treg cells. Glatman Zaretsky et al. revealed an unexpected role for Tregs in plasma cell biology. Here we determined the gene-expression profile of this new subset of FoxP3+ Treg cell, which express high levels of Treg effector molecules, similar to other non-lymphoid tissue Tregs. Gene-profiling of BM Tregs. Bone marrow Treg cells (30k) (gfp+CD25hiCD4+TCRÎ²+) (dump negative: CD19-CD8Î±-TCRÎ³Î´-CD11b-CD11c-NK1.1-Gr-1-Ter-119-) were triple-sorted from pools of two to three reporter mice (C57BL/6 Foxp3-IRES-gfp, 9 week-old males) into trizol per ImmGen SOP. RNA was amplified, labeled and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays (Expression Analysis)

2015-12-23 | E-GEOD-76264 | biostudies-arrayexpress

Transcription profile of CD103- vs CD103+ dendritic cells derived from CX3CR1+c-kit+- bone marrow cells (M1993 M2067)

Project description:We hypothesize that under homeostatic as well as inflammatory conditions circulating monocytes and/or their bone marrow-derived progenitors might contribute to the replenishment of CD103+ and CD103- DC in lymphoid and non-lymphoid compartments. To that end, bone marrow cells from CX3CR1+/gfp C57BL/6 mice were sorted as follows: lineage negative (CD3, CD19, NK1.1, Ter119, Ly6G and CD11c) CX3CR1+c-kit+. Sorted cells were further cultured in vitro under the continuous presence of GM-CSF. lin-CX3CR1+c-kit+ bone marrow cells gave rise to CD103+ and CD103- DC in vitro. To test whether CD103 might be a suitable marker that allows the differentiation of functionally distinct DC subsets generated in vitro, CD11c+CD103+ and CD11c+CD103- DC were sorted from day 5 cultures of lin-CX3CR1+c-kit+ cells and analyzed by whole mouse genome microarrays from Agilent technologies. Compared to CD103+ DC, CD103- DC displayed a strong up-regulation of transcripts for genes involved in innate immunity, whereas those involved in costimulation were down-modulated. Our data suggest distinct functional activity of these two DC subsets. Keywords: Transcriptional profiling-Cell type comparison

2008-12-17 | GSE10882 | GEO

Transcription profile of CD103- vs CD103+ dendritic cells derived from CX3CR1+c-kit+- bone marrow cells (M1993 M2067)

2008-12-17 | E-GEOD-10882 | biostudies-arrayexpress

CD38-NAD+ Immuno-Metabolic Axis Regulates Potent Anti-Tumor T Response

Project description:Spleens from the B6 mice were isolated and single cell suspension was made. CD4 T cells were purified from the splenocytes using magnetic bead separation. Briefly, Splenocytes were incubated with biotinylated antibody cocktail consisting of antibodies (Biolegend) to CD19, B220, CD11b, CD11c, NK1.1, Gr1, CD25 CD8. After a wash step, cells were incubated with streptavidin conjugated magnetic particles (BD Biosciences). After washing, CD4 T cells were isolated by applying a magnetic field and removing the untouched cells. Purified CD4 T cells were then activated with plate-bound anti-CD3 plus anti-CD28 in presence of either Th1 or Th17 or Th1/17 polarizing condition for 3 days. Total RNA from the 3 days differentiated Th1, Th17 and Th1/17 cells was isolated using mirVana miRNA isolation Kit (Invitrogen).

2017-09-13 | E-MTAB-5237 | biostudies-arrayexpress

Bulk RNA sequencing of NCR- ILC3s from the murine spleen and the small intestine

Project description:Splenic and intestinal NCR- ILC3s have been shown to be phenotypically and functional different. The goal of this study is to compare transcriptional profiles of NCR- ILC3s isolated of the murine spleen (SP) and the small intestine (SI) by RNA seq technology. Cell suspension were generated from both organs and NCR- ILC3s (CD117+, Thy1.2+, eYFP+, lin- (CD3, CD8, CD11b, CD11c, CD19, B220, Gr-1, TCRβ, TCRγδ, TER-119, NK1.1, NKp46)) were sort purified. RNA was isolated and RNA sequencing was done using Ilumina Hiseq 2500 system and NuGEN Ovation RNA Seq System V2, with biological triplicates. We provide the first comparison of transcriptional profiles of intestinal and splenic NCR- ILC3s.

2020-03-17 | GSE114749 | GEO

Comparison of NK cell developmental stages in bone marrow of Tbx21 WT versus KO mice

Project description:Bone marrow cells were isolated from wild type and Tbx21 knockout mice. iNK (Lin- NK1.1+ CD11b- CD27+) and mNK1 (Lin- NK1.1+ CD11b+ CD27+) were sorted from WT and Tbx21 KO mice. mNK2 (Lin- NK1.1+ CD11b+ CD27-) were sorted from WT mice only, since these cells are not present in Tbx21 KO mice.

2019-03-20 | GSE122874 | GEO

ScRNA-seq of dendritic cells from mouse lymph nodes

Project description:Dendritic cells were sorted from Lymph nodes and subject to single-cell sequencing. Gating strategy: CD45+ Lin- (CD3, CD19), F4/80-, CD64-, MHC+, CD11c+.

2021-04-10 | E-MTAB-10196 | biostudies-arrayexpress

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