Project description:Monocytes are key players in inflammatory processes which are triggered by lipopolysaccharide (LPS), the major outer membrane component of gram-negative bacteria. The present study in human monocytic THP-1 cells was designed in order to identify LPS-inducible genes which are down-regulated by the reduced form of CoQ10 (ubiquinol, Q10H2). For this purpose, THP-1 cells were incubated with 10 µM Q10H2 for 24 h. Subsequently, cells were stimulated for 4 h with 1µg/ml LPS and the resulting gene expression levels were determined using microarrays. 14 LPS-inducible genes were identified to be significantly (p < 0.05) down-regulated by Q10H2 pre-treatment between a factor of 1.32 and 1.65. The strongest effect of Q10H2 incubation was found for the nuclear receptor coactivator 2 gene (NCOA2). Gene Ontology (GO) terms revealed for the Q10H2-sensitive genes an involvement in e.g. signal transduction processes (CENTD1, NCOA2, PSD3, PPP2R5C), transcriptional regulation (NCOA2, POU2F1, ETV3) and cell proliferation pathways (CCDC100, EPS15). In conclusion, we provide evidence in THP-1 cells that the reduced form of CoQ10 (Q10H2) modulates LPS-induced gene expression.
Project description:RNA-Seq was carried out in order to obtain the expression profile of lipopolysaccharide (LPS)-induced transcriptome changes in PMA-differentiated human THP-1 cell line.
Project description:Monocytes are key players in inflammatory processes which are triggered by lipopolysaccharide (LPS), the major outer membrane component of gram-negative bacteria. The present study in human monocytic THP-1 cells was designed in order to identify LPS-inducible genes which are down-regulated by the reduced form of CoQ10 (ubiquinol, Q10H2). For this purpose, THP-1 cells were incubated with 10 µM Q10H2 for 24 h. Subsequently, cells were stimulated for 4 h with 1µg/ml LPS and the resulting gene expression levels were determined using microarrays. 14 LPS-inducible genes were identified to be significantly (p < 0.05) down-regulated by Q10H2 pre-treatment between a factor of 1.32 and 1.65. The strongest effect of Q10H2 incubation was found for the nuclear receptor coactivator 2 gene (NCOA2). Gene Ontology (GO) terms revealed for the Q10H2-sensitive genes an involvement in e.g. signal transduction processes (CENTD1, NCOA2, PSD3, PPP2R5C), transcriptional regulation (NCOA2, POU2F1, ETV3) and cell proliferation pathways (CCDC100, EPS15). In conclusion, we provide evidence in THP-1 cells that the reduced form of CoQ10 (Q10H2) modulates LPS-induced gene expression. Whole genome expression profiles were analysed from monocytes pre-incubated with the reduced form of CoQ10 (ubiquinol, Q10H2) before subsequent stimulation with LPS. Stimulated (+LPS) and unstimulated (-LPS) monocytes were used as positive and negative controls, respectively. For every experimental group (3 groups in total), three Affymetrix Human Genome U133 Plus 2.0 arrays were used, thus resulting in the analysis of 9 microarrays.
Project description:In order to identify patterns of gene expression associated with biological effects in THP-1 cells induced by F3, we performed a transcriptomic analysis on the THP-1 control and F3-treated THP-1 cells by oligonucleotide microarray Experiment Overall Design: 10^7 cells/mL concentrations of THP-1 cells were seeded in 100 mm dish and incubated overnight. After that, cells were treated with F3 at a final concentration of 30 ug/mL. After incubated for 6 and 24 hours, the cell pellets were collected by centrifugation at 250g for 5 min, correspondingly. Controlled samples of un-induced cells were treated in the same way with the same amount of medium.
Project description:In this study we performed single-cell transcriptome analysis of THP-1 macrophages, stimulated with high levels of free fatty acids (palmitate, PAL) typical for obese adipose tissue microenvironment or lipopolysaccharide (LPS), representing a classical stimulus activating innate immune response. Analysing full transcriptomes of individual cells, we were able to distinguish 3 macrophage transcriptional states and decipher gene regulatory pathways underlying macrophage state identity in both stimulations.
Project description:To further study the transcriptome of THP-1 human monocytes after exposure to S-Nitrosoglutathione (GSNO), we investigate whole genome microarray expression to identify genes regulated by exposure or not to GSNO. To further study the transcriptome of THP-1 human monocytes after exposure for 4 h to 50 ug / mL of S-Nitrosoglutathione-loaded polymeric Eudragit RL nanoparticles (GSNO-loaded ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 50 ug / mL of GSNO-loaded ENP. To further study the transcriptome of THP-1 human monocytes after exposure for 4 h to 200 ug / mL of empty polymeric Eudragit RL nanoparticles (empty ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 200 ug / mL of empty ENP. To further study the transcriptome of THP-1 human monocytes after exposure for 24 h to 50 ug / mL of S-Nitrosoglutathione-loaded polymeric Eudragit RL nanoparticles (GSNO-loaded ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 50 ug / mL of GSNO-loaded ENP. To further study the transcriptome of THP-1 human monocytes after exposure for 24 h to 50 ug / mL of empty polymeric Eudragit RL nanoparticles (empty ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 50 ug / mL of empty ENP. To further study the transcriptome of THP-1 human monocytes after exposure for 4 h to 50 ug / mL of empty polymeric Eudragit RL nanoparticles (empty ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 50 ug / mL of empty ENP. To further study the transcriptome of THP-1 human monocytes after exposure for 4 h to 200 ug / mL of S-Nitrosoglutathione-loaded polymeric Eudragit RL nanoparticles (GSNO-loaded ENP), we investigate whole genome microarray expression to identify genes regulated by exposure or not to 200 ug / mL of GSNO-loaded ENP. Changes in gene expression in THP-1 cells incubated without (control) or with 50 uM GSNO for 4 h, were measured. Five biological replicates were performed as controls: F_01; F_07; F_13; S_01; S_02. Four biological replicates were performed in GSNO exposed cells: S_13; S_14; S_15; S_16. Changes in gene expression in THP-1 cells incubated without (control) or with 50 ug / mL of GSNO-loaded ENPs (300 nm) for 4 h were measured. Five biological replicates were performed as controls: F_01; F_07; F_13; S_01; S_02. Three biological replicates were performed in 50 ug / mL of GSNO-loaded ENP exposed cells: S_06; S_07; S_08. Changes in gene expression in THP-1 cells incubated without (control) or with 200 ug / mL of empty ENPs (300 nm) for 4 h were measured. Five biological replicates were performed as controls: F_01; F_07; F_13; S_01; S_02. Three biological replicates were performed in 200 ug / mL of empty ENP exposed cells: S_17; S_19; S_20. Changes in gene expression in THP-1 cells incubated without (control) or with 50 ug / mL of GSNO-loaded ENPs (300 nm) for 24 h were measured. Five biological replicates were performed as controls: F_04; F_10; F_16; S_03; S_04. Four biological replicates were performed in 50 ug / mL of GSNO-loaded ENP exposed cells: S_09; S_10; S_11; S_12. Changes in gene expression in THP-1 cells incubated without (control) or with 50 ug / mL of empty ENPs (300 nm) for 24 h were measured. Five biological replicates were performed as controls: F_04; F_10; F_16; S_03; S_04. Three biological replicates were performed in 50 ug / mL of empty ENP exposed cells: F_05; F_11; F_17. Changes in gene expression in THP-1 cells incubated without (control) or with 50 ug / mL of empty ENPs (300 nm) for 4 h were measured. Five biological replicates were performed as controls: F_01; F_07; F_13; S_01; S_02. Three biological replicates were performed in 50 ug / mL of empty ENP exposed cells: F_02; F_08; F_14. Changes in gene expression in THP-1 cells incubated without (control) or with 200 ug / mL of GSNO-loaded ENPs (300 nm) for 4 h were measured. Five biological replicates were performed as controls: F_01; F_07; F_13; S_01; S_02. Four biological replicates were performed in 200 ug / mL of GSNO-loaded ENP exposed cells: S_21; S_22; S_23; S_24.
Project description:Direct comparison of the genome-level expression patterns of THP-1 cells exposed to either LPS or heat shock; Peripheral blood mononuclear cells (PBMC) serve a sentinel role allowing the host to efficiently sense and adapt to the presence of danger signals. Herein we have directly compared the genome-level expression patterns (microarray) of human PBMC (THP-1 cells) subjected to one of two canonical danger signals, heat shock and lipopolysaccharide (LPS). Based on sequential expression and statistical filters, and in comparison to control cells, we found that 3,988 genes were differentially regulated in THP-1 cells subjected to LPS stress, and 2,921 genes were differentially regulated in THP-1 cells subjected to heat shock stress. Venn analyses demonstrated that the majority of differentially regulated genes (greather than or equal to 70%) were uniquely expressed in response to one of the two danger signals. Functional analyses demonstrated that the two danger signals induced expression or repression of genes corresponding to unique pathways, molecular functions, biological processes, and gene networks. In contrast, there were 184 genes that were commonly upregulated by both stress signals, and 430 genes that were commonly downregulated by both stress signals. Interestingly, the 184 commonly upregulated genes corresponded to a gene network broadly related to inflammation, and more specifically to chemokine signaling. These data demonstrate that the mononuclear cell responses to the canonical stress signals, heat shock and LPS, are highly divergent. However, there is a heretofore unrecognized common pattern of gene network expression corresponding to chemokine-related biology. The data also serve as a reference database for investigators in the field of stress signaling. Experiment Overall Design: In vitro exposure of THP-1 cells to control conditions, LPS (1 micogram/ml for 4 hrs), or heat shock (43 deg C for 1 hour, followed by a 4 hour recovery at 37 deg C)
Project description:Analysis of the transcriptome of THP-1 cells upon Huh7 cell-derived ectosomes incubation. Monocytic THP-1 cells were incubated with or without ectosomes (10 mg/ml) derived from scramble sequence transfection (Scramble Ecto) or PKM2 knocking down (PKM2 KD Ecto) Huh7 cells for 24 h. Results provide a novel insight into monocyte differentiation.
Project description:This study investigated the global effect of carbon monoxide (CO) gas (250 ppm) on LPS- induced gene expression in THP-1 cells. CO predominantly suppresses LPS-induced gene response. Of the CO-suppressed genes, 18% were transcription factors and most others were cytokines, chemokines and immune response genes. Sequence analysis B binding siteskappafound that 81% of the gene promoters have putative NF- These results suggest that CO inhibits LPS-induced inflammatory B signaling pathwayresponse through regulating NF- The microarrays were performed on THP-1 cells, a human monocytic cell line. Cells were treated with or without LPS (1 ug/ml) in presence or absence of carbon monoxide (250 ppm) for 1 h. Total RNA was isolated, reverse transcribed, labeled and hybridized to oligonucleotide microarrays.
Project description:So far, we have found phorbol 12-myristate 13-acetate (PMA) induced ubiquitin specific peptidase (USP) 2b isoform in myeloid leukemia cell lines such as HL60, THP-1, and U937. HL60, THP-1, and U937 undergoes differentiation into macrophage-like cells after stimulation with phorbol ester. To explore molecular function of USP2 in macrophages especially during lipopolysaccharide(LPS) response, we assess expression profiles of HL60-derivatives continuously expressing shRNA for USP2 and control shRNA.