Project description:B1 cells account for the majority of B cell population in the peritoneal cavity, and are essential for the innate immune responses and maintaining the homeostasis. The origin of the B1 cells and how to form the B1 cells pool in the postnatal life remain unknown. And the heterogeneity of B1 cells can largely affect the functions of B1 cells. Until now, nobody has performed the single cell RNA-seq of peritoneal B cells. In order to reveal the characteristics of peritoneal B cells, we have performed the scRNA-seq and scBCR-seq of the peritoneal B cells of mouse from different stages.
Project description:The B-cell receptor (BCR) enables individual B cells to identify diverse antigens, including bacterial and viral proteins. While advances in RNA-seq have enabled high throughput profiling of transcript expression in single cells, the unique task of assembling the full-length heavy and light chain sequences from single cell RNA-sequencing (scRNA-seq) in B cells has been largely unstudied. We developed a new software tool, BASIC, which allows investigators to use scRNA-seq for assembling BCR sequences at single cell level. To demonstrate the utility of our software, we subjected single B cells from a human donor to scRNA-seq, assembled the full-length heavy and the light chains, and experimentally confirmed these results by using single cell primer based nested PCRs and Sanger sequencing.
Project description:In Beclinf/f-Lyz2cre mice, we observed proinflammatory activation of tissue-resident peritoneal macrophages. These mice were resistent to lethal bacterial infection with Listeria. It is important to understand how the peritoneal immunity is affected by Beclin deletion in myeloid cells. To characterize the immune cells in the peritoneum, we performed single cell RNA sequencing of total peritoneal cells from Beclinf/f-Lyz2cre and littermate control Beclinf/f mice. The single cell RNA-seq analyses revealed profound changes in both the myeloid compartment and bystander cells.
Project description:To define the transcriptomic landscape of human mesothelial cells in response to PD, we analyzed single cell transcriptomes including cells dissociated from normal peritoneum (hernia surgery, n = 3), and peritoneal cells from effluent of short-term PD patients (PD less than 2 weeks, n = 6) and from long-term PD patients (PD more than 6 years, n = 4) ,we demonstrate that mesothelial cells develop hyperglycolysis, a metabolic alteration that is correlated with the development of peritoneal fibrosis.
Project description:Purpose:Flow cytometric analysis showed that after 48h of culture with T cell- and Ag-derived stimuli, GCBC alter their TF expression and begin to differentiate. However, only a subset of cells undergoes this process, for reasons that are unclear. To better define the nature of this heterogeneity, and to further define the differences between IL-21R/CD40 vs BCR/CD40 signals, we performed single cell RNA-seq (scRNA-seq) of 48h GCBC cultures with either of these two stimuli or with the combination of IL-21R/CD40 and BCR stimulation. Methods: we performed scRNA-seq using 10X Genomics Chromium system . Results: Single cell analysis demonstrated surprisingly little overlap between the clusters derived from IL-21/⍺-CD40 stimulation and those derived from BCR/⍺-CD40 ligation, thus establishing at molecular and cellular detail how distinct these two types of selection signals are for GCBC