Project description:Human liver myeloid cells are imperfectly defined, but it is broadly agreed that cells of stellate appearance in situ, expressing the markers CD11b and CD68, are the liver’s resident macrophages, termed Kupffer cells. Recent investigations using single cell RNA sequencing and unsupervised clustering algorithms suggest there are two populations of cells with the characteristics of tissue macrophages in human liver. We therefore analyzed dissociated human liver tissue using the markers CD11b and CD68 to define Kupffer cells and found within this population two subsets that differ in their expression of multiple surface markers. These subsets were FACS-sorted based on CD32 expression, and gene expression analysis identified them with human liver myeloid cell subsets defined by two independent single cell RNA sequencing studies. These two subsets differed in the expression of genes associated with T cell activation and immunosuppression, suggesting distinct roles in T cell tolerance.
Project description:Separation of B cells has been historically important in discovering their functional relevance, particularly in relation to infection, immune disorders and vaccination. Traditional use of phenotypic markers often poses problems in distinguishing heterogeneous populations such as the Double Negative (DN, CD19+CD27-IgD-) cells. B cells represent a small subset of PBMCs; this represents challenges to use bottom-up approaches such as single-cell transcriptomics in defining B cell subpopulations. In this study we therefore used the 10X single-cell RNAseq platform on B cell populations already defined by FACS sorting (Transitional, CD19+CD27-IgD+CD10+; Naïve, CD19+CD27-IgD+CD10-; Classical Memory, CD19+CD27+IgD-; IgM Memory, CD19+CD27+IgD+; and DN). These data match known phenotypes to transcriptionally defined B cell subpopulations, and provide a reference atlas for researchers interested in better defining B cell subsets in their data.
Project description:Previous reports have defined three subsets of mouse NK cells on the basis of the expression of CD27 and CD11b. The developmental relationship between these subsets was unclear. To address this issue, we evaluated the overall proximity between mouse NK cell subsets defined by CD27 and CD11b expression using pangenomic gene expression profiling. The results suggest that CD27+CD11b-, CD27+CD11b+ and CD27-CD11b+ correspond to three different intermediates stages of NK cell development.
Project description:It is known that natural killer (NK) cells are a heterogeneous population of functionally distinct NK cell subsets. Here we report on different genomic, phenotypic and functional properties of human NK cell subsets derived from peripheral blood, thymus and bone marrow. NK cell subpopulations were defined via expression of CD56 and CD16.