Project description:CD138+ B220- plasma cells were sorted from bone marrow and B220+ CD23+ mature follicular B cells were sorted from the spleens. Plasma cells were sorted from C57BL/6 mice 7 days after boosting with antigen, with mice first primed with an i.p. injection of KLH/IFA followed by boost at day 21 with KLH/PBS i.p. Mature B cells were sorted from antigen-naïve C57BL/6 mice. We compared expression profiles of plasma cells and mature B cells to identify differentially expressed transcripts.
Project description:CD138+ B220- plasma cells were sorted from bone marrow and B220+ CD23+ mature follicular B cells were sorted from the spleens. Plasma cells were sorted from C57BL/6 mice 7 days after boosting with antigen, with mice first primed with an i.p. injection of KLH/IFA followed by boost at day 21 with KLH/PBS i.p. Mature B cells were sorted from antigen-naïve C57BL/6 mice. We compared expression profiles of plasma cells and mature B cells to identify differentially expressed transcripts. Seven days after boosting with KLH, plasma cells were enriched by CD138-PE/anti-PE magnetic bead enrichment and then sorted to purity. Affymetrix microarrays were performed on 50,000-70,000 plasma cells and 2,000,000-3,000,000 naïve B cells.
Project description:Long-lived plasma cells (LLPCs) develop under the help of follicular helper T (Tfh) cells and reside mainly in the bone marrow. However, these cells are unusually abundant in the spleen of several autoimmune models including K/BxNsf mice, yet their pathogenic impact remains unknown. To investigate a previously unappreciated role of splenic LLPCs, we sorted splenic plasma cells (PCs) from K/BxNsf and K/BxN mice, corresponding to LLPCs and conventional short-lived PCs, respectively, and compared their transcriptomes.
Project description:Long-lived plasma cells (LLPCs) develop under the help of follicular helper T (Tfh) cells and reside mainly in the bone marrow. However, these cells are unusually abundant in the spleen of several autoimmune models including K/BxNsf mice, yet their pathogenic impact remains unknown. To investigate a previously unappreciated role of splenic LLPCs, we sorted splenic plasma cells (PCs) from K/BxNsf and K/BxN mice, corresponding to LLPCs and conventional short-lived PCs, respectively, and compared their transcriptomes. The B220+CD138+ fraction from K/BxNsf and their control K/BxN mice were sorted by FACSariaIII.
Project description:To understand tissue resident features of memory CD4+ and CD8+ T lymphocytes of the bone marrow and/or spleen according to expressing or not the tissue retention marker CD69, we performed whole transcriptome profiling of ex vivo antigen-specific CD69+ and CD69- memory CD4+ T cells isolated from bone marrow and spleen, and ex vivo CD69+ and CD69- memory CD8+ T cells isolated from bone marrow.
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.
Project description:Two mice in which luciferase-positive LOUCY ETP-ALL cells were xenografted were treated with either vehicle or ABT-199 (50 mg/kg) for 11 days. Afterwards, single cells suspensions were made from bone marrow and spleen of both mice for 10x Genomics scRNA-seq. Following quality control, a total of 13,681 spleen cells and 11,442 bone marrow cells of the control mouse and 15,955 spleen cells and 9,443 bone marrow cells of the ABT 199 treated mouse were processed for further analysis. Our data show that ABT-199 triggers transcriptional changes in T-cell differentiation genes in leukemic cells obtained from the spleen microenvironment. These results are in line with our previous findings that mature typical T-ALL are resistant to ABT-199.
Project description:Maturation, age, tissue localization and functional capacity all drive neutrophil heterogeneity. In mouse models of cancer, we found that Ly6GInt neutrophils were abundant, and their frequency correlated with metastatic potential. Only Ly6G surface expression was found to accurately identify neutrophil maturity by flow cytometry across models, with other markers being model specific. Using several stimuli, we found that Ly6GInt neutrophils are bone fide ‘immature neutrophils’ with reduced immune regulatory and adhesion capacity. The spleen is a site of neutrophil production in homeostasis and cancer. Strikingly, we found that neutrophils mature and undergo post-mitotic transit faster in the spleen than in the bone marrow with unique transcriptional profiles for splenic Ly6GInt and Ly6GHi neutrophils. We propose that developmental origin is critical in neutrophil identity and postulate that neutrophils that develop in the spleen supplement the bone marrow by providing an intermediate more mature reserve before emergency haematopoiesis.
Project description:The mature peripheral B cell compartment consists of follicular (FO) and marginal zone (MZ) B cells, which mature from transitional B cells in the spleen and mount T-dependent and T-independent antibody responses, respectively. TACI (Tnfrsf13b) is a member of the TNF-receptor superfamily expressed on mature and transitional B cells, with highest expression on MZ B cells and plasma cells, which is activated by the cytokines BAFF and APRIL. BAFF also stimulates the related receptor BAFFR, which provides signals that are essential for mature B-cell survival. Previous studies of TACI-/- mice have observed that TACI-deficiency leads to an expansion of mature B cells, and therefore it has long been thought that TACI is a negative regulator of mature B-cell survival. However, TACI-/- mice also have elevated levels of BAFF, therefore the B-cell hyperplasia phenotype in TACI-/- mice is most likely a cell-extrinsic effect due to increased BAFF-BAFFR signalling in TACI-/- B cells. To investigate the cell-intrinsic role of TACI, we generated mouse mixed bone marrow chimeras from TACI-/- and WT mice and studied the effect of TACI loss in vivo, without the confounding factor of increased BAFF. Here, we have analysed the transcriptomes of TACI KO and TACI WT marginal zone and follicular B cells by RNAseq.
Project description:In this study, we explored gene expression profiles of CD34-CLEC12Ahi pre-basophils isolated from the bone marrow as well as CD34-CLEC12Alo mature basophils from the bone marrow and CD34+ basophil progenitors. We revealed that the gene expression of mature basophils isolated from the bone marrow and spleen resembled each other whereas pre-basophils and mature basophils showed distinct gene expression profiles.
