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.
Project description:CD34+ cells were isolated from leukopheresis preparations by Miltenyi positive selection kits for CD34 cells and cultured in stemPro medium+ supplement 2 culture conditions were compared, G1 = IL-3 at 5 ng/ml for the 17 days of culture or G3 = IL-3 at 5 ng/ml + SCF (stem cell factor) at 4 ng/ml + Flt3L at 8 ng/ml
Project description:Basophils are the least common granulocytes which represent less than 1% of peripheral blood leukocytes. Recent development of analytical tools for basophils enables us to understand their critical roles in allergic reactions and protection from parasitic infections. Nevertheless, the differentiation trajectory of basophils remains unclear. We have identified previously-unappreciated pre-basophil subpopulation which encompasses previously-defined basophil precursors (BaPs) in a series of experiments including scRNA-seq analysis (deposited to GSE206589). In this study, we explored gene expression profiles of pre-basophils and mature basophils activated by antigen/IgE-stimulation or IL-3-stimulation. We have revealed that IL-3 activated pre-basophils displayed distinct gene expression profiles from antigen/IgE- or IL-3-stimulated mature basophils, suggesting the unuque property of pre-basophils.
Project description:To investigate the mechanisms by which C/EBPa drives basophil differentiation and maintains basophil identity, we examined whether or not C/EBPa promotes basophil molecular programming and simultaneously represses mast cell molecular programming. We performed genome-wide gene expression profiling on basophils and mast cells and found that 6798 genes were shared by mast cells and basophils; 2033 genes were expressed 2-10 (log2 1-3.3)-fold higher in basophils (differentially expressed in basophils); and 413 genes were expressed greater than 10 (log2 3.3)-fold in basophils (highly expressed in basophils). On the other hand, we found 569 genes were expressed 2-10 (log2 -1 to -3.3) fold higher in mast cells and 171 genes were highly expressed in mast cells [greater than 10 fold (log2 -3.3)]. We treated purified basophils prepared from Cebpaf/f RosaYFP/creER mice and Cebpa+/+ RosaYFP/creER control mice with or without 4HT treatment for five days. Gene expression in the treated basophils was analyzed using microarray analysis. Overall, deletion of C/EBPa in basophils resulted in a reduction of mRNA expression for 248 genes and led to an increase in mRNA expression for 255 genes. The majority of the C/EBPa-regulated genes were either differentially or highly expressed in basophils or mast cells. In this study, we compared gene expression in basophils and mast cell and identified genes which specifically expressed in basophils and mast cells. By using Cebpa conditional knock out mice, we identified Cebpa regulated genes in basophils.
Project description:In this study, we explored gene expression profiles of mature and immature basophils from either IL-3-elicited or TSLP-elicited BMBAs. We revealed that TSLP-elicited mature basophils displayed gene expression profile similar to that of IL-3-elicited mature rather than immature basophils.
Project description:CD34+ progenitor cells were cultured for 0 or 21 days in StemPro medium + supplement + IL-3 at 5 ng/ml Basophils, eosinophils and plasmacytoid dendritic cells were purified to near homogeneity and lysed for miRNA and mRNA. Basophils were purified by negative selection (StemSep Abs, miltenyi columns), eosinophils by StemSep Abs and pDC by positive selection after removing monocytes. The method starting with a 2-step Percoll gradient that put eosinophils in the pellet, basophils in the middle layer and pDC in the upper layer.
Project description:Innate lymphoid cells (ILCs) were generated in vitro from CD34+ hematopoietic progenitors derived from umbilical cord blood, and RNA sequencing was performed on CD45+ LineageLD- cells to assess transcriptional identities of lineages generated