Inflammation-induced chemokine expression in uveal melanoma cell lines stimulates monocyte chemotaxis
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE55983 | GEO | 2014/07/30
SECONDARY ACCESSION(S): PRJNA241516
REPOSITORIES: GEO
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Project description:Purpose: Uveal melanoma (UM) is the most common primary intraocular tumor in adults and the presence of infiltrating leucocytes is associated with a poor prognosis. Little is known how infiltrating leucocytes influence the tumor cells. The purpose of this study was to investigate the effect of activated T cells on the expression of chemotactic cytokines in UM cells. Furthermore, we examined the ability of stimulated UM cells to attract monocytes. Methods: We used an in vitro co-culture system in which UM cell lines and T cells were cultured together, but separated by a membrane. UM gene expression was quantified using a microarray. Protein expression in the supernatant was quantified with ELISA or cytometric bead array. For the monocyte migration assay, a trans-well plate was used. Results: Gene-expression analysis of UM cell lines showed that co-culture with activated T cells resulted in an upregulation of chemokines such as CXCL8, CXCL9, CXCL10, CXCL11, CCL2, CCL5, vascular endothelial growth factor (VEGF), intracellular adhesion molecule 1 (ICAM1) and granulocyte-macrophage colony-stimulating factor (GM-CSF). The upregulation of these molecules was confirmed at the protein level. This increase of chemokines coincided with an increased chemotactic capacity of the supernatant towards monocytes. Conclusions: Cytokines derived from activated T cells shifted the UM cell-transcriptome towards a more inflammatory state, including upregulation of several chemokines, which led to an increased migration of monocytes. Therefore, UM cells might actively participate in generating a tumor- promoting inflammatory microenvironment. One replicate for each cell lines (92.1, Mel 270 and Mel 290) was analyzed before and after co-culture with activated T cells.
2014-07-30 | E-GEOD-55983 | biostudies-arrayexpress
Project description:Chronic liver diseases as non-alcoholic steatohepatitis (NASH)-induced cirrhosis are characterized by an increasing accumulation of stressed, damaged, or dying hepatocytes. Hepatocyte damage triggers the activation of resident immune cells, such as Kupffer cells (KC), as well as the recruitment of immune cells from the circulation towards areas of inflammation. After infiltration, monocytes differentiate into monocyte-derived macrophages (MoMF) which are functionally distinct from resident KC. We herein aim to compare in vitro signatures of polarized macrophages and activated hepatic stellate cells (HSC) with ex vivo derived disease signatures from human NASH. Secondly, we investigate the effects of the secretome of primary human monocytes, macrophages and NK cells on HSC activation, as in particular monocytes and macrophages have often been linked to liver fibrosis progression. IL-4 and IL-13 treatment induced TGF-β1 secretion by macrophages. However, the supernatant transfer did not induce HSC activation. Interestingly, PMA-activated macrophages showed a strong induction of the fibrosis response genes COL10A1 and CTGF, while supernatant of IL-4/IL-13 treated monocytes induced the upregulation of COL3A1 in HSC. Supernatant of PMA-activated NK cells had the strongest effect on COL10A1 induction in HSC, while IL-15 stimulated NK cells reduced the expression of COL1A1 and CTGF. These data indicate that other factors, aside the well-known cytokines and chemokines, are potentially stronger contributors to activation of HSCs and induction of a fibrotic response, indicating a more diverse and complex role of monocytes, macrophages and NK cells in liver fibrosis progression.
2024-03-20 | GSE228086 | GEO
Project description:Chronic liver diseases as non-alcoholic steatohepatitis (NASH)-induced cirrhosis are characterized by an increasing accumulation of stressed, damaged, or dying hepatocytes. Hepatocyte damage triggers the activation of resident immune cells, such as Kupffer cells (KC), as well as the recruitment of immune cells from the circulation towards areas of inflammation. After infiltration, monocytes differentiate into monocyte-derived macrophages (MoMF) which are functionally distinct from resident KC. We herein aim to compare in vitro signatures of polarized macrophages and activated hepatic stellate cells (HSC) with ex vivo derived disease signatures from human NASH. Secondly, we investigate the effects of the secretome of primary human monocytes, macrophages and NK cells on HSC activation, as in particular monocytes and macrophages have often been linked to liver fibrosis progression. IL-4 and IL-13 treatment induced TGF-β1 secretion by macrophages. However, the supernatant transfer did not induce HSC activation. Interestingly, PMA-activated macrophages showed a strong induction of the fibrosis response genes COL10A1 and CTGF, while supernatant of IL-4/IL-13 treated monocytes induced the upregulation of COL3A1 in HSC. Supernatant of PMA-activated NK cells had the strongest effect on COL10A1 induction in HSC, while IL-15 stimulated NK cells reduced the expression of COL1A1 and CTGF. These data indicate that other factors, aside the well-known cytokines and chemokines, are potentially stronger contributors to activation of HSCs and induction of a fibrotic response, indicating a more diverse and complex role of monocytes, macrophages and NK cells in liver fibrosis progression.
2024-03-20 | GSE228087 | GEO
Project description:Purpose: To determine to which extend inflammatory cytokines affect chemokine secretion by primary human choroidal melanocytes (HCMs), their capacity to attract monocytes and if HCMs are able to influence the proliferation of activated T cells. Methods: Primary cultures of HCMs were established from eyes of thirteen donors. HCMs were stimulated with IFN-γ and TNF-α or with supernatant from activated T-cells (T-cell conditioned media (TCM)). Gene expression analysis was performed using microarrays. Protein levels were quantified with ELISA or cytometric bead array. Supernatants of HCMs were assessed for the capability to attract monocytes in a trans-well plate. Proliferation of activated T-cells was assessed in a direct co-culture with HCMs by a [3H]-thymidine incorporation assay. Results: Stimulation of HCMs with TCM or IFN-γ and TNF-α resulted in increased expression and secretion of CXCL8, CXCL9, CXCL10, CXCL11, CCL2, CCL5 and intercellular adhesion molecule 1. Vascular endothelial growth factor and monocyte migration inhibitory factor were constitutively expressed without changes in response to pro-inflammatory cytokines. Supernatants derived from unstimulated cultures of ten HCM donors induced a high initial level of monocyte migration, which decreased upon stimulation with either TCM or IFN-γ and TNF-α. The supernatants from three HCM donors initially showed a low level of monocyte attraction which increased following exposure to pro-inflammatory cytokines. Direct co-culture of HCMs with T-cells resulted in inhibition of T-cell proliferation. Conclusion: These results show that normal and activated HCMs are immunologically active by secreting chemokines, and that HCMs are able to attract monocytes in addition to inhibit T-cell proliferation.
2016-12-21 | GSE70762 | GEO
Project description:To gain a comprehensive understanding of type I IFN or SARS-CoV-2 activated pDCs priming macrophages in regulating TLR4 meidated gene expression, we treated primary human blood monocytes with 30ng/ml IFNα or the supernatant of SARS-CoV-2 activated pDCs for 24 h and followed by 2 ng/ml LPS for 3 h. Cells were harvested and performed transcriptomic analysis via RNA-seq. We found that IFNα and supernatant of SARS-CoV-2 activated pDCs had similar effects on priming macrophages to induce massive inflammatory gene expression mediated by LPS.
2022-08-05 | GSE182572 | GEO
Project description:Differential response of bovine mammary epithelial cells to Staphylococcus aureus or Escherichia coli agonists of the innate immune system. Mastitis caused by Escherichia coli and Staphylococcus aureus is a major pathology of dairy cows. To better understand the differential response of the mammary gland to these two pathogens, we stimulated bovine mammary epithelial cells (bMEC) with either E. coli crude lipopolysaccharide (LPS) or with S. aureus culture supernatant (SaS) to compare the transcriptomic profiles of the initial bMEC response (3 and 6 h of exposure to agonists). By using HEK 293 cells transfected with human pattern recognition receptors, the LPS preparation was found to stimulate TLR2 and TLR4 but not TLR5, Nod1 or Nod2, whereas SaS stimulated TLR2. Biochemical analysis revealed that lipoteichoic acid, protein A and alpha-hemolysin were all present in SaS, and bMEC were found to be responsive to each of these molecules. Transcriptome profiling by a microarray and confirmation by RT-qPCR revealed an innate immune response which was common to both LPS and SaS. However, LPS induced expression of a significant higher number of genes and the fold changes were of greated magnitude than those induced by SaS. Overall, the analysis of microarray data suggests that the activation pathways and the early chemokine and cytokine production preceded the defense and stress responses. Chemokines were among the most up-regulated genes, in particular ELRCXC chemokines that target neutrophils. A major differential response was the activation of the type I IFN pathway by LPS but not by SaS. This was in accordance with the much stronger up-regulation of Cxcl10, Ccl5 and Nos2 by LPS than by SaS. The higher upregulation of chemokines that target mononuclear leucocytes (CXCL10, CCL2, CCL5 and CCL20) by LPS than by SaS is likely to be related to the differential activation of the type I IFN pathway, and could induce a different profile of the initial recruitment of leucocytes. It is noteworthy that at the protein level, secretion of TNF-alpha and IL-1-beta was not induced by either stimulus. These results suggest that the response of MEC to diffusible bacterial stimuli is able to contribute to the onset of the response with differential leucocyte recruitment but does not account directly for the differential production of major pro-inflammatory cytokines. Transcriptional profiling of bovine mammary epithelial cells (Holstein breed) comparing untreated control with mammary epithelial cells (MECs) stimulated by Staphylococcus aureus (SA) versus Escherichia coli lipopolysaccharide (LPS). 16 microarrays (2 stimuli * 2 times * quadruplicate) in a two-color dye-swap experimental design. Untreated cells served as the control. One replicate per array. Log2-intensity of each dye was analyzed separately => 32 samples.
2014-01-11 | E-GEOD-47599 | biostudies-arrayexpress
Project description:Chemokines are chemotactic cytokines that control the migratory patterns and positioning of immune cells. Many chemokines have been associated with systemic autoimmune diseases that have chronic IFN signature. We now describe that a series of chemokines, including CXCL4, CXCL10, CXCL12 and CCL5, can superinduce type I IFN (IFN-I) by TLR9-activated plasmacytoid DCs (pDCs), independently of their respective known chemokine receptors. Mechanistically, we show that chemokines such as CXCL4 mediate transcriptional and epigenetic changes in pDCs, mostly targeted to the IFN-I pathways. We also describe that chemokines physically interact with DNA to form nanoparticles which promote clathrin-mediated cellular uptake and delivery of DNA in the early endosomes of pDCs. Using two mouse models of skin inflammation, we observed the presence of CXCL4 associated with DNA in vivo. These data thus reveal a non-canonical role for chemokines to modulate TLR signaling, and extend our understanding on the mechanism associated with the chronic expression of IFN-I by pDCs in autoimmune diseases.
2022-05-24 | GSE202333 | GEO
Project description:Differential response of bovine mammary epithelial cells to Staphylococcus aureus or Escherichia coli agonists of the innate immune system. Mastitis caused by Escherichia coli and Staphylococcus aureus is a major pathology of dairy cows. To better understand the differential response of the mammary gland to these two pathogens, we stimulated bovine mammary epithelial cells (bMEC) with either E. coli crude lipopolysaccharide (LPS) or with S. aureus culture supernatant (SaS) to compare the transcriptomic profiles of the initial bMEC response (3 and 6 h of exposure to agonists). By using HEK 293 cells transfected with human pattern recognition receptors, the LPS preparation was found to stimulate TLR2 and TLR4 but not TLR5, Nod1 or Nod2, whereas SaS stimulated TLR2. Biochemical analysis revealed that lipoteichoic acid, protein A and alpha-hemolysin were all present in SaS, and bMEC were found to be responsive to each of these molecules. Transcriptome profiling by a microarray and confirmation by RT-qPCR revealed an innate immune response which was common to both LPS and SaS. However, LPS induced expression of a significant higher number of genes and the fold changes were of greated magnitude than those induced by SaS. Overall, the analysis of microarray data suggests that the activation pathways and the early chemokine and cytokine production preceded the defense and stress responses. Chemokines were among the most up-regulated genes, in particular ELRCXC chemokines that target neutrophils. A major differential response was the activation of the type I IFN pathway by LPS but not by SaS. This was in accordance with the much stronger up-regulation of Cxcl10, Ccl5 and Nos2 by LPS than by SaS. The higher upregulation of chemokines that target mononuclear leucocytes (CXCL10, CCL2, CCL5 and CCL20) by LPS than by SaS is likely to be related to the differential activation of the type I IFN pathway, and could induce a different profile of the initial recruitment of leucocytes. It is noteworthy that at the protein level, secretion of TNF-alpha and IL-1-beta was not induced by either stimulus. These results suggest that the response of MEC to diffusible bacterial stimuli is able to contribute to the onset of the response with differential leucocyte recruitment but does not account directly for the differential production of major pro-inflammatory cytokines.
2014-01-11 | GSE47599 | GEO
Project description:Blood monocytes serve as the first line of host defense and are equipped to recognize and respond to infection by triggering an immune-inflammatory response. While most information on these cells comes from in vitro studies in humans or in vivo studies in mice, little is known about monocytes under human disease conditions. We investigated the role of monocytes during sepsis and its resolution in humans. A transcriptomal and functional analysis of blood monocytes from patients during gram negative sepsis and at recovery was performed. Monocytes from sepsis patients showed upregulation of a large number of pro-inflammatory genes and cytokines/chemokines, consistent with an ongoing systemic inflammation. However, these cells showed impairment to ex vivo endotoxin (LPS) challenge, displaying a quantitative decrease in the number of LPS-inducible genes. Moreover, they downregulated the expression of several pro-inflammatory cytokine/chemokine genes, activation marker genes and transcription factors associated with monocyte/macrophage activation, upon ex vivo LPS stimulation. Functionally, they downregulated expression of inflammatory cytokines/chemokines and antigen presentation-related molecules and functions. In contrast, genes and functions related to phagocytosis, anti-microbial activity and tissue remodeling where remained unaffected or even enhanced . Collectively, our observations suggest a genetic and functional re-programming of these cells during human sepsis progression. Understanding the molecular mechanisms which regulate this re-programming will allow to devise strategies which could modulate the response of these cells and hence, disease progression. Blood monocytes from gram-negative sepsis patients during sepsis (Sepsis) and following their recovery (Recovery/Basal) as well as healthy donor (control) were isolated. Thereafter, these cells were treated ex vivo with or without LPS for 3h and analysed for transcriptomic study.
2014-05-14 | E-GEOD-46955 | biostudies-arrayexpress
Project description:EVs were isolated from primary human aortic endothelial cells (ECs) (+/- IL-1b activation), quantified, and analysed by miRNA transcriptomics and proteomics. Compared to quiescent ECs, activated ECs increased EV release, with miRNA and protein cargo that were related to atherosclerosis pathways. RNA sequencing of EV-treated monocytes and vascular smooth muscle cells (VSMCs) revealed that EVs from activated ECs altered pathways that were pro-inflammatory and atherogenic.
2024-01-02 | GSE231715 | GEO