Project description:Our objective was to investigate epigenomic and transcriptomic changes in J774 macrophages after incubation with talc and titanium dioxide particles. This dataset comprises the results of reduced representation bisulfite sequencing (RRBS) of the J774 cell DNA 24 hours after exposure to 10 ug/well of fine-sized talc or titanium dioxide particles in vitro with or without 2 ug/mL 17-b estradiol.
Project description:Bone-marrow macrophages polarized to M2 phenotype are immunosuppressive. Interestingly, treatment with whole-glucan particles converts M2 macrophages to M1 phenotype with an anti-tumor phenotype. In this study, the effect of WGP treatment for 6 hours on the gene expression of M2 macrophages was assessed.
Project description:Triglyceride-rich lipoproteins and their remnants contribute to atherosclerosis, possibly by carrying remnant cholesterol and/or by exerting a pro-inflammatory effect on macrophages. Nevertheless, little is known about how macrophages process triglyceride-rich lipoproteins. We show that uptake by macrophages of VLDL-sized emulsion particles is dependent on the enzyme lipoprotein lipase via its C-terminal domain. Subsequent internalization of VLDL-triglycerides by macrophages is carried out by caveolae-mediated endocytosis, followed by hydrolysis by lysosomal acid lipase. STARD3 is required for the transfer of lysosomal fatty acids to the ER for lipid storage, while NPC1 likely is involved in promoting the extracellular efflux of fatty acids. Our data provide novel insights into how macrophages process VLDL-derived triglycerides and suggest that macrophages have the remarkable capacity to excrete internalized triglycerides as fatty acids.
Project description:Viral antigens can activate phagocytes inducing inflammation but the mechanisms are barely explored. This study aimed to investigate the capability of viral oligomeric proteins of different structure to induce inflammatory response in macrophages. Human THP-1 cell line was used to prepare macrophages which were treated with filamentous nucleocapsid-like particles (NLPs) of paramyxoviruses and spherical virus-like particles (VLPs) of human polyomaviruses. The effects of viral proteins on cell viability, pro-inflammatory cytokines’ production and formation of NLRP3 inflammasome components, ASC specks, were investigated. Filamentous NLPs did not induce inflammation markers while spherical VLPs mediated inflammatory response followed by NLRP3 inflammasome activation. Inhibitors of cathepsins and K+ efflux decreased IL-1β levels and cell death indicating a complex inflammasome activation process. Similar activation pattern was observed in primary human macrophages treated with VLPs. Single cell RNAseq analysis of THP-1 cells revealed several cell activation states characterized by high expression of inflammation-related genes. This study provides new insights into interaction of viral proteins with innate immune cells and suggests that structural properties of oligomeric proteins may define cell activation pathways.
Project description:Long term use of titanium implants generates wearing particles off. Titanium nanoparticles (TiNP) favor a pro-inflammatory macrophage polarization (M1) and lower the tolerogenic activation (M2). Our previous Affymetrix data showed that TiNP upregulate GDF15 and downregulate STAB1 expression in macrophages. GDF15 is a cytokine that regulates energy expenditure and fibrosis, and is endocytosed by the scavenger receptor Stabilin-1. Our study aim was to investigate the effect of TiNP on GDF15 and STAB1 expression, on GDF15 secretion and on endocytosis efficiency in macrophages. We used the model system of primary human macrophages derived from CD14+ monocytes. After 6 days of incubation with TiNP, we quantified GDF15 and STAB1 expression, and GDF15 secretion in INF-γ stimulated (M1) and IL-4 stimulated (M2) macrophages using RT-PCR and ELISA, respectively. Flow cytometry assessed the effect of TiNP on endocytosis based on the uptake of fluorescently labelled acLDL, a known ligand of stabilin-1, and MS-1, an antibody directed against the extracellular portion of stabilin-1 in M1 and M2. Our results revealed a strong up-modulating effect of TiNP on GDF15 expression and secretion in all subtypes of macrophages. In contrast, stabilin-1 expression was significantly suppressed by TiNP. acLDL and MS-1 uptake by macrophages was also significantly suppressed under the treatment with TiNP. Our findings highlight a consistent ex vivo stimulatory effect of TiNP on GDF15 production and inhibitory effect on STAB1 expression and Stabilin-1-mediated endocytosis. Our results suggest an important implication of TiNP on macrophage healing functions.
Project description:In this study gene expression of monocyte-derived macrophages (MDM) from chronic obstructive pulmonary disease (COPD) patients and healthy subjects was investigated. MDM were treated with LPS, a combination of fine TiO2 and ultrafine Printex90 particles, or remained untreated. Experiment Overall Design: MDM of 13 COPD patients and 13 healthy subjects were incubated for four hours with LPS (10ng/ml), a combination of fine TiO2 and ultrafine Printex90 (32ug/ml each) or remained untreated. Cells were harvested, counted, and total RNA was isolated (phenol-chloroform extraction) for each subject individually. Total RNA of the 13 individuals from each group (COPD, healthy) and incubation (untreated, LPS, particles) were pooled and hybridized on a seperate array.
Project description:The toxicity and toxicogenomics of selected anatase and rutile nanoparticles (NP) and bulk titanium dioxide (TiO2) particles were evaluated in the soil nematode Caenorhabditis elegans. Results indicated that bulk or nano-TiO2 particles were slightly toxic to soil nematode C. elegans, as measured by reproduction EC50 values ranging from 4 to 32 mg/L. Whole-genome microarray results indicated that the regulation of glutathione-S-transferase gst-3, cytochrome P450 cypp33-c11, stress resistance regulator scl-1, oxidoreductase wah-1, and embryonic development pod-2 genes were significantly affected by nano-sized and bulk TiO2 particles. More specifically, it was determined that anatase particles exerted a greater effect on metabolic pathways, whereas rutile particles had a greater effect on developmental processes. The up-regulation of the pod-2 gene corroborated the phenotypic effect observed in the reproduction test. Our results demonstrated that C. elegans is a good genomic model for nano-TiO2 toxicity assessment.
Project description:We previously demonstrated by genomic and bioinformatical approaches that human macrophage (MΦ) activation is best described by a spectrum model (Xue et al, Immunity, 2014). MΦ integrate exogenous input signals on transcriptional level in a unique fashion to generate specific functional programs, enabling the plasticity in disease-related pathophysiologies. Such versatile responsiveness requires fast changes of transcription mediated by transcriptional regulators (TRs) or epigenomic changes. To better understand the principles of this regulation during human MΦ activation, we assessed histone modifications including H3K4me1, H3K4me3, H3K27me3, and H3K27Ac by ChIP-sequencing allowing us to characterize the functional state of promoters (active, poised, repressed) and enhancers (active, inactive, intermediate). Using transcriptome data from our MΦ spectrum model, we generated a co-regulation network of all TRs. Next, we overlaid epigenomic information and transcriptional changes of major TRs over time onto the TR network. We observed that input signals like IFNγ or TNFα induce a specific network of TRs that are transcriptionally regulated themselves, the combination of regulated TRs changes over time with a boost of transcriptional regulation of dozens of TRs 4 to 12 hrs post input signal exposure, almost all TRs within the network show active promoters, even if the TR itself is not expressed, and similar results are obtained for enhancers with open or at least intermediated states. These findings strongly suggest that in MΦ, the TR-defined cellular â??switch panelâ?? is always accessible thereby allowing MΦ to quickly respond to the diverse input signal repertoire from the environment. Epigenetic analysis of promoter and enhancer sites in primary human macrophage subtypes and correlation to RNA-seq expression data