Project description:NK cells from healthy donors were isolated from peripheral blood and stimulated overnight with IL-12/15/18 to induce a memory-like phenotype. After 7 days of culture, the phenotype and degranulation potential of CIML NK cells was characterized in two distinct NK cell subsets: CD16+/CD56+ and CD16−/CD56. Finally, NK cell subsets were purified using fluorescence-activated cell sorting (FACS) and subjected to high-throughput multiplexed quantitative proteomics.
Project description:ATAC-seq experiments were performed to map the open chromatin regions of human, chimpanzee and macaque iAstrocytes. This experiment allowed to compare evolutionary changes according to chromatin activity.
Project description:Uterine NK cells (uNK cells) form a distinct immune cell population in the endometrium and decidua. Here, we FACS-sorted KIR-CD39-,KIR+CD39- and KIR+CD39+ uNK cells from decidual samples.
Project description:Identification of nuclear CSF-1R, H3K4me1 and H3K4me3 localization on chromatin in human primary monocytes and modification of this localization during monocyte differentiation into macrophage induced by 100ng/mL CSF-1 during 6 hours (1 donor) or 72h (3 donors). Identification of EGR1 chromatin localization in human primary monocytes (3 donors) Comparison of nuclear CSF-1R chromatin localization in monocytes from 1 healthy donor and 2 chronic myelomonocytic leukemia patients.
Project description:A map of global Irx3 binding sites during early stages of adipogenic differentiation. C57BL/gNJ (B6N) mice were fed a SDS maintenance chow diet, and at the age of 6-10 weeks, gWAT and iWAT was isolated and primary preadipocytes were isolated. For each depot, cells from 6-12 mice were pooled. Isolated cells were cultured until confluence in 2D and induced to differentiate 2 days post confluence by a standard adipogenic cocktail. One day before (day -1) and one day after (day 1) induction of differentiation, cells were fixated by formaldehyde, treated with protease inhibitors, snap-frozen and shipped to Active Motif for ChIP-seq analysis. Reads were aligned to mouse genome (mm10) using Bowtie2 (version 2.3.4.3), with duplicates and low quality mapping reads (<30) removed via samtools. Peak files were annotated using the R package ChIPseeker (version 1.22.0). Log2 ChIP-over-input tracks for each alignment were generated using deepTools bamCompare (version 3.1.2), and blacklisted regions from Encode were excluded. Bam files were submitted to Macs2 (version 2.1.1) for peak calling. A q-value of 1E-4 and 10X enrichment was used as threshold.
Project description:Identification of EGR1 chromatin localization in human primary monocytes (2 donors), in human primary monocytes from 2 CMML patients with TET2 truncating mutations (high VAF) and in human primary monocytes from 1 CMML patients with TET2 truncating mutations with low VAF.
Project description:Innate lymphoid cells (ILCs) are part of the innate immune cell family. Three different subsets of ILCs, ILC1s, ILC2s and ILCPs can be identified in human peripheral blood. Based on their expression of transcription factors and cytokines, they are considered as being the innate counterparts of CD4 T helper subsets, namely Th1s, Th2s and Th17s. However, ILCs and Th cells have different roles in immunity. Therefore, we compared the transcriptomes of sorted ILC1s, ILC2s, ILCPs, Th1s, Th2s and Th17s from the peripheral blood of three different donors. RNA sequencing of ILC and Th subsets revealed differences in the expression of tens to hundreds of genes. These genes are involved in cell trafficking, innate activation and inhibitory functions. ILC and Th cell subsets also differ in their expressions of long-non coding RNAs.
Project description:Embryonic cardiomyocytes possess the plasticity to choose between atrial and ventricular fates. For a limited window of time, the transcription factor COUP-TFII (Nr2f2) sufficiently and essentially confers the atrial identity through direct and indirect regulation of nearly half of chamber specific genes. Examination of COUP-TFII binding sites in embryonic artia
Project description:CD56neg CD16+ Natural Killer (NK) cells have been reported to expand in chronic diseases and acute myeloid leukemia (AML). However, their biological role is still unclear. Using bulk RNA-sequencing, we compared gene expression profile of CD56neg CD16+ NK cells with conventional CD56bright, CD56dim CD16- and CD56dim CD16+ NK cells from blood samples of HV and AML patients. RNA-seq unveiled that CD56neg CD16+ NK cells were mature circulating NK cells with functional capacities. Taken together, our results suggest that CD56neg CD16+ NK cells are a relevant target for future NK-cell-based immunotherapies.