Project description:We want to define the differences in gene expression changes between neonatal and adult myeloid dendritic cells upon stimulation with TLR7/8. These differences may point to specific signaling pathways which, in turn, may account for the known differential immune response of babies to vaccination or infection when compared to adults.
Project description:We want to define the differences in gene expression changes between neonatal and adult plasmacytoid dendritic cells upon stimulation with TLR7/8. These differences may point to specific signaling pathways which, in turn, may account for the known differential immune response of babies to vaccination or infection when compared to adults.
Project description:Previous studies have shown that purified dendritic cells (DCs) have cell-intrinsic, age-dependent differences in their response to TLR stimulation. To delineate which aspects of the age-dependent difference in innate immunity are cell intrinsic vs extrinsic, we searched for global differences to TLR7/8 stimulation in purified adult vs neonatal DC populations. We hypothesize that very few selected cell intrinsic differences in gene expression of key immune genes between these 2 age groups exist, and the bulk would be cell extrinsic differences. The results show that there are age-dependent differences in expression of several key genes involved in the immune response at baseline already. Upon stimulation, we identified a substantially larger fraction of age-dependent differentially expressed genes in conventional than plasmacytoid DCs. Bioinformatics analyses indicate that important immune pathways were significantly differentially expressed in DC subsets between the 2 age groups. Total RNA was isolated from purified human conventional and plasmacytoid dendritic cells from 6 adult and 6 cord blood donors that were stimulated with 3M-003 at a final concentration of 5 uM for 1 and 6 hr.
Project description:Myeloid dendritic cells (mDC) were isolated from antiretroviral therapy (ARV)-treated and untreated people living with HIV (PLWH), and from HIV uninfected Individuals. The results indicate that mDC are altered in genes expression from PLWH. Some RNA transcriptional changes are not completely restored by ARV. This provides more data on myeloid cells, an understudied cell type, and alterations in PLWH.
Project description:Previous studies have shown that purified dendritic cells (DCs) have cell-intrinsic, age-dependent differences in their response to TLR stimulation. To delineate which aspects of the age-dependent difference in innate immunity are cell intrinsic vs extrinsic, we searched for global differences to TLR7/8 stimulation in purified adult vs neonatal DC populations. We hypothesize that very few selected cell intrinsic differences in gene expression of key immune genes between these 2 age groups exist, and the bulk would be cell extrinsic differences. The results show that there are age-dependent differences in expression of several key genes involved in the immune response at baseline already. Upon stimulation, we identified a substantially larger fraction of age-dependent differentially expressed genes in conventional than plasmacytoid DCs. Bioinformatics analyses indicate that important immune pathways were significantly differentially expressed in DC subsets between the 2 age groups.
Project description:With increasing age, the ability of the immune system to protect against recurring infections or to control chronic infections erodes. The objective of the current study was to identify gene expression signatures in elderly CD4 T cell responses Vβ2+ CD4 memory T cells from four 20-35 and four 70-75 year-old individuals were stimulated with toxic shock syndrome toxin (TSST) presented by myeloid dendritic cells (mDC) derived from young adults. Gene expression was examined at 16, 40 and 72 h after stimulation using Affymetrix arrays.
Project description:We studied the role of Notch2 signaling in Ly6Chi monocyte cell fate during TLR7-induced acute inflammation. To characterize the gene expression changes involved in monocyte differentiation, we subjected monocyte subsets from peripheral blood of wt and myeloid Notch2 mutant mice after Sham or IMQ treatment to RNA-sequencing and gene expression analysis. We found that Cell-intrinsic Notch2 and TLR7-Myd88 pathways independently and synergistically promote Ly6Clo patrolling monocyte development from Ly6Chi monocytes under inflammatory conditions. At the same time TLR7 stimulation in the absence of functional Notch2 signaling promotes resident tissue macrophage gene expression signatures in monocytes and ectopic differentiation of Ly6Chi monocytes into macrophages and dendritic cells. Thus, Notch2 is a master regulator of Ly6Chi monocyte cell fate and inflammation in response to TLR signaling.
Project description:To investigate the expression of glycosyltransferases in dendritic cells and the changes in expression associated with maturation. RNA preparations from 5 different immature and mature dendritic cells subsets, with and without LPS from 2 different donors (20 different samples); (iDC, IL-10 iDC, Dexamethason iDC, Vitamin D3 iDC, im. macrophage), (mDC, IL-10 mDC, Dexamethason mDC, Vitamin D3 mDC, m. macrophage) were sent to both Microarray Core (E) and Core(C). The RNA was purified on an RNeasy Column, amplified, labeled, and hybridized to the Glycov3 microarrays.