Project description:In this study, we designed an in vitro approach to help us understand how extracellular components participate in human B cell functional plasticity and how this network might be involved in auto-immune diseases. Two activated T cell subsets were used to expose a common pool of B cells to distinct, controlled microenvironments. We showed that the same B cell population could experience two possible opposite evolutions, to either helper or suppressor functions, in a flexible manner. These cells were thus referred to as effector B cells (Eff B) or suppressor B cells (Supp B). We thus analyzed their different transcriptional programs by RNA sequencing.
Project description:The investigators hypothesize that the treatment of metastatic colorectal cancer (mCRC) patients with the combination of alpha-type-1-polarized dendritic cell (αDC1) vaccines and tumor-selective chemokine modulation (CKM) will promote the infiltration of vaccination-induced CD8+ CTLs to tumor lesions and subsequently tumor regression with improved patient survival.
Project description:Human stem cell-derived hepatocyte-like cells (HLCs) offer an attractive platform to study liver disease and to test drugs. However, despite their advantages over other hepatocyte culture systems, HLCs lack several in vivo characteristics including cell polarity, which is critical for proper hepatocyte biology. We report a stem cell-based differentiation protocol that uses transwell filters to generate columnar polarized HLCs with clearly defined basolateral and apical membranes separated by tight junctions. Unlike conventional HLCs, polarized HLCs secrete cargo directionally, similar to hepatocytes in vivo. This novel system provides a powerful tool to study hepatocyte biology, disease mechanisms, genetic variation, and drug metabolism in a more physiologically relevant setting.
Project description:The loss of apicobasal polarity during the epithelial-to-mesenchymal transition (EMT) is a hall-mark of cancer and metastasis. The key feature of this polarity in epithelial cells is the subdivision of the plasma membrane into apical and basolateral domains, with each orchestrating specific in-tra- and extracellular functions. Epithelial transport and signaling capacities are thought to be determined largely by the quality, quantity and nanoscale organization of proteins residing in these membrane domains, the apicobasal surfaceomes. Despite its implications for cancer, drug uptake and infection, our current knowledge of how the polarized surfaceome is organized and maintained is limited. Here we used chemoproteomic surfaceome scanning to establish proteo-type maps of apicobasal surfaceomes and reveal quantitative distributions of i.a. surface proteas-es, phosphatases and tetraspanins as potential key regulators of polarized cell functionality. We show further that tumor-suppressor PTEN regulates polarized surfaceome architecture and un-cover a potential role in collective cell migration. Our differential surfaceome analysis provides a molecular framework to elucidate polarized protein networks regulating epithelial functions and PTEN-associated cancer progression.
Project description:Using stable isotope labelling by amino acids in cell culture (SILAC)-based quantitative proteomics on purified centrosomes from resting, non-activated non-polarized, and activated polarized mouse B cells, we established a list of proteins differentially associated with the centrosome between resting and polarized states.
Project description:Unstimulated (M0), M1-polarized (GM-CSF, LPS, IFNγ-stimulated), and M2-polarized (M-CSF, IL-4-stimulated) canine blood-derived macrophages were generated in vitro and investigated for differences in their transcriptome to create a basis for future investigations upon the role of macrophage polarization in dogs, a species, which has emerging importance for translational research.
Project description:We reported exosome-guided phenotype switches between M1- and M2-polarized BMDMs. M1- or M2-polarized BMDMs were successfully reprogrammed to M2- or M1-phenotype via the treatment of exosomes obtained from M2- or M1-polarized BMDMs. In this uploaded information, the exosomes from M1- and M2-polarized BMDMs were analyzed by high-throughput sequencing.
Project description:To understand the molecular programs driving Th17 heterogeneity and ATAC CD4 T cell fate, we profiled in vitro polarized non-pathogenic Th17, pathogenic Th17, Th1, and Treg cells using ATAC-seq.
Project description:This project involves proteome characterization of healthy monocytes polarized (activated) with synovial fluid (SF) versus serum from patients with juvenile idiopathic arthritis (JIA). The objective is to investigate the influence of SF on monocytes at the proteome level, using serum polarized monocytes as a control. Monocytes were isolated from peripheral blood mononuclear cells of healthy donors following negative selection and were subsequently polarized overnight with 20% of a pool of either serum or SF. They were subsequently detached and used for proteome analysis.
