Project description:RNAseq of germinal center B cells with different Blimp-1-GFP reporter level. Sorted germinal center populations were defined by their expression of Blimp-1-GFP reporter and the cell surface marker CD138 before they were further split into dark zone or light zone / dark zone like or light zone like cells based on their expression of the surface marker CXCR4 and CD83. We used RNAseq to find genes differentially regulated in various Blimp-1-GFP expressing germinal center populations from the spleen at day 10 after intraperitoneal immunisation with sheep red blood cells.
Project description:RNAseq of germinal center B cells (B220+IgDlowCD95+GL7+) with different Blimp-1-GFP reporter level in the presence of TfH cells or 72 hr after their depletion. Sorted germinal center populations were defined by their expression of Blimp-1-GFP reporter and the cell surface marker CD138. Analysed mice were bone marrow chimeras generated with a bone marrow mixture from CD4-cre+ Rosa26-loxP-stp-loxP-DTR Blimp-1GFP/+ (10%) and Rag1-/- OTII (90%) mice. In these mice TfH cells can be depleted by the injection of human diphteria toxin (DT).
Project description:Productive B cell responses are critical to protect a host from infection. The spleen and lymph nodes are populated by resting follicular B cells, which can enter germinal centers upon antigen encounter. Once in the germinal center, B cells migrate between the dark and light zones, where they undergo somatic hypermutation and selection, respectively. While germinal center B cells have been studied, an intense molecular understanding of these cells/subsets (and the differences between them) is lacking.
Project description:WT-AID-GFP+ and SFR KO-AID-GFP+ mice were immunized with NP(18)OVA+ alum, on Day 9, germinal center B cells (AID-GFP+) were sorted for total RNA
Project description:A range of cellular interactions and cytokines influence immunoglobulin class switch recombination (CSR), which occurs both in extrafollicular antibody responses and germinal centers. This study addresses where CSR occurs during extrafollicular responses, allowing identification of the cellular interactions and cytokine-producing cells that influence CSR in vivo. The response in mice of B cells specific for 4-hydroxy-3-nitrophenyl-acetyl (NP) to NP-Ficoll was analyzed. Immunohistology and flow cytometry confirm responding NP-binding cells move to the outer T zone of the spleen and start dividing there on the second day. During the third day they move either to follicles as germinal center founding cells, or extrafollicular foci as plasmablasts. Analysis of expression of AID, Bcl-6 and Blimp-1 mRNA in single NP-binding cells indicates a proportion of T zone B blasts express AID transiently without Bcl-6 and these cells are shown to undergo CSR. By contrast Blimp-1+ cells emerging on the third day and were universally AID- and hence unable to initiate CSR. Bcl-6 mRNA without protein, expressed by 40% of naïve NP-binding cells, is rapidly downregulated. Bcl-6 and AID are coexpressed by germinal centre founding cells only on moving to follicles during the third day after immunization. Keywords: TaqMan Low Density Array
Project description:Germinal center B cells were isolated from human tonsil tissue and crosslinked. ChIP was performed on two distinct pools of germinal center cells, each obtained from 3-5 donors. The experiment includes two biological replicates (germinal center cell pools from different donors). ChIP was performed on both pools and subject to library preparation and sequencing. Input DNA was sequenced for both pools.
Project description:Given the tumor suppressing function of miR-15a/16-1 cluster, we studied its role in the germinal center B-cells that give rise to most lymphoid malignancies.