Project description:Numerous studies have shown in mouse that identity and function of macrophage populations are imprinted by their tissue of residence. By contrast, the properties of human tissue macrophages remain poorly understood. Here, we characterized human tonsil macrophages and identified 3 macrophage subsets with distinct phenotype, ontogeny and function. Using RNA-seq analysis, we found that CD36hi macrophages were transcriptionally related to monocytes, while CD36lo macrophages showed features of embryonic origin and CD36int macrophages had a mixed profile. Using scRNA-seq profiling of naïve and inflamed tonsils from non-human primates, we showed that monocyte engraftment did not pre-exist an immune challenge.
Project description:B cells from human tonsil and blood were sorted using flow cytometry. The human samples were processed immediately ex-vivo using markers for known B cell subsets.
Project description:Numerous studies have shown in mouse that identity and function of macrophage populations are imprinted by their tissue of residence. By contrast, the properties of human tissue macrophages remain poorly understood. Here, we characterized human tonsil macrophages and identified 3 macrophage subsets with distinct phenotype, ontogeny and function. Using RNA-seq analysis, we found that CD36hi macrophages were transcriptionally related to monocytes, while CD36lo macrophages showed features of embryonic origin and CD36int macrophages had a mixed profile. Using scRNA-seq profiling of naïve and inflamed tonsils from non-human primates, we showed that monocyte engraftment did not pre-exist an immune challenge.
Project description:Follicular helper CD4 T (Tfh) cells provide B cells with signals that are important for the generation of high-affinity Abs and immunological memory and, therefore, are critical for the protective immunity elicited by most human vaccines. In this study we sought to define the gene expression signature of bona fide GC Tfh and Tfh cells. The CD4+ T cell subsets CD45RO+CXCR5-, CD45RO+CXCR5int (Tfh cells), and CD45RO+CXCR5hi (GC Tfh cells) were isolated from 6 tonsil samples for gene expression analysis.
Project description:To characterize a novel human ILC2 subset found in peripheral blood, RNAseq was performed. Other ILC subsets from blood and tonsil were also included.
Project description:Macrophages are a heterogeneous population of immune cells that play central roles in a broad range of biological processes, including the resolution of inflammation. Although diverse macrophage subpopulations have been identified, the characterization and functional specialization of certain macrophage subsets in inflamed tissues remain unclear. Here we uncovered a key role of specific macrophage subsets in tissue repair using proteomics, bioinformatics and functional analyses. We isolated two hepatic monocyte-derived macrophage subpopulations: Ly6ChiCX3CR1lo macrophages and Ly6CloCX3CR1hi macrophages during distinct phases of acute liver injury and employed label-free proteomics approach to profile the proteome of these cells. We found that the wound healing- and endocytosis-related proteins were specifically enriched in Ly6CloCX3CR1hi macrophages. Intriguingly, 12/15-lipoxygenase (Alox15), the most strongly up-regulated protein in Ly6CloCX3CR1hi macrophages, was identified as a specific marker for these macrophages. In co-culture systems, Ly6CloCX3CR1hi macrophages specifically induced hepatocyte proliferation. Furthermore, selective depletion of this population in CD11b-diphtheria toxin receptor mice significantly delayed liver repair. Overall, our studies shed light on the functional specialization of distinct macrophage subsets in the resolution of inflammation.
Project description:We used an IL4-capture assay followed by FACS sorting, to isolate IL4-secreting TFH cells from a human tonsil and compared their transcriptomic profiles with CXCR5hi PD1hi IL4-negative tonsillar TFH cells and IL4-producing CXCR5neg non-TFH cells (TH2 cells). Our studies validate the notion of functionally distinct TFH subsets and identify genes that are specifically expressed in and define the human IL-4 secreting TFH cell subset.