Transcriptome profiling of hiPSC-derived LSECs with nanoCAGE

2020 ◽  
Vol 16 (2) ◽  
pp. 138-146 ◽  
Author(s):  
Mathieu Danoy ◽  
Stéphane Poulain ◽  
Yuta Koui ◽  
Yannick Tauran ◽  
Benedikt Scheidecker ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Transcriptome Profiling ◽  
Liver Endothelial Cells

As a response to the challenge of the in vitro maintenance of liver endothelial cells, we have used hiPSCs-derived LSECs and have measured their performance by comparing them to their primary counterpart using the nanoCAGE technology.

scholarly journals Characterization of the interaction both in vitro and in vivo of tissue-type plasminogen activator (t-PA) with rat liver cells. Effects of monoclonal antibodies to t-PA

1992 ◽  
Vol 284 (2) ◽  
pp. 545-550 ◽  
Author(s):  
M Otter ◽  
J Kuiper ◽  
R Bos ◽  
D C Rijken ◽  
T J van Berkel
Keyword(s):  
Endothelial Cells ◽  
Mannose Receptor ◽  
Liver Cells ◽  
Tissue Type ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator ◽  
Liver Endothelial Cells ◽  
Parenchymal Cells

The interaction of 125I-labelled tissue-type plasminogen activator (125I-t-PA) with freshly isolated rat parenchymal and endothelial liver cells was studied. Binding experiments at 4 degrees C with parenchymal cells and endothelial liver cells indicated the presence of 68,000 and 44,000 high-affinity t-PA-binding sites, with an apparent Kd of 3.5 and 4 nM respectively. Association of 125I-t-PA with parenchymal cells was Ca(2+)-dependent and was not influenced by asialofetuin, a known ligand for the galactose receptor. Association of 125I-t-PA with liver endothelial cells was Ca(2+)-dependent and mannose-specific, since ovalbumin (a mannose-terminated glycoprotein) inhibited the cell association of t-PA. Association of 125I-t-PA with liver endothelial cells was inhibited by anti-(human mannose receptor) antiserum. Anti-(galactose receptor) IgG had no effect on 125I-t-PA association with either cell type. Degradation of 125I-t-PA at 37 degrees C by both cell types was inhibited by chloroquine or NH4Cl, indicating that t-PA is degraded lysosomally. in vitro experiments with three monoclonal antibodies (MAbs) demonstrated that anti-t-PA MAb 1-3-1 specifically decreased association of 125I-t-PA with the endothelial cells, and anti-t-PA Mab 7-8-4 inhibited association with the parenchymal cells. Results of competition experiments in rats in vivo with these antibodies were in agreement with findings in vitro. Both antibodies decreased the liver uptake of 125I-t-PA, while a combination of the two antibodies was even more effective in reducing the liver association of 125I-t-PA and increasing its plasma half-life. We conclude from these data that clearance of t-PA by the liver is regulated by at least two pathways, one on parenchymal cells (not galactose/mannose-mediated) and another on liver endothelial cells (mediated by a mannose receptor). Results with the MAbs imply that two distinct sites on the t-PA molecule are involved in binding to parenchymal cells and liver endothelial cells.

Uptake and Degradation of Tissue Plasminogen Activator in Rat Liver

1988 ◽  
Vol 59 (03) ◽  
pp. 474-479 ◽  
Author(s):  
Monica Einarsson ◽  
Bård Smedsrød ◽  
Håkan Pertoft
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Rat Liver ◽  
Liver Cells ◽  
Terminal Phase ◽  
Tissue Plasminogen ◽  
Liver Endothelial Cells ◽  
Parenchymal Cells

SummaryThe mechanism of uptake of tissue plasminogen activator (tPA) in rat liver was studied. Radio-iodinated tPA was removed from the circulation after intravenous administration in a biphasic mode. The initial half life, t1/2(α), and the terminal phase, t1/2(β), were determined to be 0.5 min and 7.5 min, resp. Separation of the liver cells by collagenase perfusion and density centrifugation, revealed that the uptake per cell was two to three times higher in the non-parenchymal cells than in the parenchymal cells.Endocytosis of fluorescein isothiocyanate-labelled or 125I-labelled tPA was studied in pure cultures of liver cells in vitro. Liver endothelial cells and parenchymal cells took up and degraded tPA. Endocytosis was more efficient in liver endothelial cells than in parenchymal cells, and was almost absent in Kupffer cells.Competitivb inhibition experiments showing that excess unlabelled tPA could compete with the uptake and degradation of 125I-tPA, suggested that liver endothelial cells and parenchymal cells interact with the activator in a specific manner. Endocytosis of trace amounts of 125I-tPA in cultures of liver endothelial cells and parenchymal cells was inhibited by 50% in the presence of 19 nM unlabelled tPA. Agents that interfere with one or several steps of the endocytic machinery inhibited uptake and degradation of 125I-tPA in both cell types.These findings suggest that 1) liver endothelial cells and parenchymal cells are responsible for the rapid hepatic clearance of intravenously administered tPA; 2) the activator is taken up in these cells by specific endocytosis, and 3) endocytosed tPA is transported to the lysosomes where it is degraded.

scholarly journals Endocytosis of formaldehyde-treated serum albumin via scavenger pathway in liver endothelial cells

1984 ◽  
Vol 218 (1) ◽  
pp. 81-86 ◽  
Author(s):  
R Blomhoff ◽  
W Eskild ◽  
T Berg
Keyword(s):  
Endothelial Cells ◽  
Low Density Lipoprotein ◽  
Scavenger Receptor ◽  
Density Lipoprotein ◽  
Low Density ◽  
Surface Culture ◽  
Rat Liver Cells ◽  
Liver Endothelial Cells ◽  
Isolated Liver

Denatured or modified proteins (including albumin and low-density lipoprotein) are catabolized in vitro via scavenger receptors. We have studied the distribution of formaldehyde-denatured albumin in rat liver cells after intravenous injection of tracer doses of the protein. At 12 min after injection, most of the formaldehyde-denatured albumin (about 70% of the injected dose) was recovered in liver endothelial cells. Furthermore, isolated liver endothelial cells in suspension and in surface culture took up formaldehyde-denatured albumin by receptor-mediated endocytosis. Our data indicate that the scavenger receptor in liver is mainly located on the endothelial cells. Implications for the catabolism of low-density lipoproteins are discussed.

An in vitro model of warm hypoxia–reoxygenation injury in human liver endothelial cells

2012 ◽  
Vol 178 (1) ◽  
pp. e35-e41 ◽  
Author(s):  
Neal R. Banga ◽  
K. Raj Prasad ◽  
J. Lance Burn ◽  
Shervanthi Homer-Vanniasinkam ◽  
Anne Graham
Keyword(s):  
Endothelial Cells ◽  
Human Liver ◽  
In Vitro Model ◽  
Reoxygenation Injury ◽  
Liver Endothelial Cells ◽  
Vitro Model ◽  
Hypoxia Reoxygenation

scholarly journals TLR1/2 Ligand–Stimulated Mouse Liver Endothelial Cells Secrete IL-12 and Trigger CD8+T Cell Immunity In Vitro

2013 ◽  
Vol 191 (12) ◽  
pp. 6178-6190 ◽  
Author(s):  
Jia Liu ◽  
Min Jiang ◽  
Zhiyong Ma ◽  
Kirsten K. Dietze ◽  
Gennadiy Zelinskyy ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
T Cell ◽  
Mouse Liver ◽  
Cd8 T Cell ◽  
T Cell Immunity ◽  
Cell Immunity ◽  
Liver Endothelial Cells

scholarly journals New Scavenger Receptor-Like Receptors for the Binding of Lipopolysaccharide to Liver Endothelial and Kupffer Cells

1998 ◽  
Vol 66 (11) ◽  
pp. 5107-5112 ◽  
Author(s):  
Marijke van Oosten ◽  
Erika van de Bilt ◽  
Theo J. C. van Berkel ◽  
Johan Kuiper
Keyword(s):  
Endothelial Cells ◽  
Kupffer Cells ◽  
Binding Sites ◽  
Oxidized Ldl ◽  
Scavenger Receptor ◽  
Scavenger Receptors ◽  
Liver Endothelial Cells ◽  
Parenchymal Cells

ABSTRACT Lipopolysaccharide (LPS) is cleared from the blood mainly by the liver. The Kupffer cells are primarily responsible for this clearance; liver endothelial and parenchymal cells contribute to a lesser extent. Although several binding sites have been described, only CD14 is known to be involved in LPS signalling. Among the other LPS binding sites that have been identified are scavenger receptors. Scavenger receptor class A (SR-A) types I and II are expressed in the liver on endothelial cells and Kupffer cells, and a 95-kDa receptor, identified as macrosialin, is expressed on Kupffer cells. In this study, we examined the role of scavenger receptors in the binding of LPS by the liver in vivo and in vitro. Fucoidin, a scavenger receptor ligand, significantly reduced the clearance of 125I-LPS from the serum and decreased the liver uptake of 125I-LPS about 40%. Within the liver, the in vivo binding of 125I-LPS to Kupffer and liver endothelial cells was decreased 72 and 71%, respectively, while the binding of 125I-LPS to liver parenchymal cells increased 34% upon fucoidin preinjection. Poly(I) inhibited the binding of 125I-LPS to Kupffer and endothelial cells in vitro 73 and 78%, respectively, while poly(A) had no effect. LPS inhibited the binding of acetylated low-density lipoprotein (acLDL) to Kupffer and liver endothelial cells 40 and 55%, respectively, and the binding of oxidized LDL (oxLDL) to Kupffer and liver endothelial cells 65 and 61%, respectively. oxLDL and acLDL did not significantly inhibit the binding of LPS to these cells. We conclude that on both endothelial cells and Kupffer cells, LPS binds mainly to scavenger receptors, but SR-A and macrosialin contribute to a limited extent to the binding of LPS.

scholarly journals Oncostatin M induces IL-33 expression in liver endothelial cells in mice and expands ST2+CD4+lymphocytes

2015 ◽  
Vol 309 (7) ◽  
pp. G542-G553 ◽  
Author(s):  
Muhammad Imran Arshad ◽  
Pierre Guihard ◽  
Yannic Danger ◽  
Gregory Noel ◽  
Jacques Le Seyec ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fluorescent Protein ◽  
Vascular Endothelial Cells ◽  
Oncostatin M ◽  
Dependent Manner ◽  
Liver Sinusoidal Endothelial Cells ◽  
Liver Endothelial Cells ◽  
Green Fluorescent

Interleukin (IL)-33 is crucially involved in liver pathology and drives hepatoprotective functions. However, the regulation of IL-33 by cytokines of the IL-6 family, including oncostatin M (OSM) and IL-6, is not well studied. The aim of the present study was to determine whether OSM mediates regulation of IL-33 expression in liver cells. Intramuscular administration in mice of an adenovirus encoding OSM (AdOSM) leads to increase in expression of OSM in muscles, liver, and serum of AdOSM-infected mice compared with control mice. The increase of circulating OSM markedly regulated mRNA of genes associated with blood vessel biology, chemotaxis, cellular death, induction of cell adhesion molecules, and the alarmin cytokine IL-33 in liver. Steady-state IL-33 mRNA was upregulated by OSM at an early phase (8 h) following AdOSM infection. At the protein level, the expression of IL-33 was significantly induced in liver endothelial cells [liver sinusoidal endothelial cells (LSEC) and vascular endothelial cells] with a peak at 8 days post-AdOSM infection in mice. In addition, we found OSM-stimulated human microvascular endothelial HMEC-1 cells and human LSEC/TRP3 cells showed a significant increase in expression of IL-33 mRNA in a dose-dependent manner in cell culture. The OSM-mediated overexpression of IL-33 was associated with the activation/enrichment of CD4+ST2+cells in liver of AdOSM-infected mice compared with adenovirus encoding green fluorescent protein-treated control mice. In summary, these data suggest that the cytokine OSM regulates the IL-33 expression in liver endothelial cells in vivo and in HMEC-1/TRP3 cells in vitro and may specifically expand the target CD4+ST2+cells in liver.

scholarly journals Uptake and degradation in vivo and in vitro of laminin and nidogen by rat liver cells

1989 ◽  
Vol 261 (1) ◽  
pp. 37-42 ◽  
Author(s):  
B Smedsrød ◽  
M Paulsson ◽  
S Johansson
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Basement Membranes ◽  
Normal Individuals ◽  
Rat Liver Cells ◽  
Order Of Magnitude ◽  
Liver Endothelial Cells ◽  
Parenchymal Cells

Laminin antigens are known to be present in the blood of normal individuals. In the present study we have investigated the fate of laminin-related molecules in the circulation. After intravenous injection in rats, the native laminin-nidogen complex, as well as the separated proteins, were rapidly eliminated from the blood (half-lives 2-10 min) by the liver. The large laminin fragments E1 and E8 (Mr 400,000 and 280,000 respectively), which contain the major cell-adhesion-promoting activities of laminin, were also cleared from the blood mainly by the liver, but the rate of uptake was an order of magnitude lower for these fragments than for laminin. This indicates that the uptake of laminin did not occur via cell-adhesion receptors. The endothelial cells of liver were the most important cell type in the uptake of laminin-nidogen complex, nidogen, laminin and fragment E1, whereas the parenchymal cells were responsible for more than 50% of the uptake of E8 in the liver. Studies in vitro with cultured liver endothelial cells and parenchymal cells demonstrated that the ligands were endocytosed and degraded independently of plasma factors. The results reveal that the level of laminin antigens in blood is a very complex parameter. It is not only dependent on the turnover of basement membranes, but also on the degree of degradation of the material released into the blood and on the functional state of the liver, particularly the liver endothelial cells.

scholarly journals Transcriptome Profiling of Toxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin

2020 ◽  
Vol 8 (6) ◽  
pp. 842 ◽  
Author(s):  
Mohammad S. R. Harun ◽  
Mica Taylor ◽  
Xing-Quan Zhu ◽  
Hany M. Elsheikha
Keyword(s):  
Endothelial Cells ◽  
Toxoplasma Gondii ◽  
Protein Biosynthesis ◽  
Transcriptome Profiling ◽  
Global Gene Expression ◽  
Wnt Signalling ◽  
Signalling Pathway ◽  
Response To Treatment ◽  
Wnt Signalling Pathway

Central to the progression of cerebral toxoplasmosis is the interaction of Toxoplasma gondii with the blood-brain barrier (BBB) endothelial cells. In the present work, we tested the hypothesis that inhibition of Wnt pathway signalling by the monovalent ionophore monensin reduces the growth of T. gondii infecting human brain microvascular endothelial cells (hBMECs) or microglial cells. The anti-parasitic effect of monensin (a Wnt signalling inhibitor) on the in vitro growth of T. gondii tachyzoites was investigated using two methods (Sulforhodamine B staining and microscopic parasite counting). The monensin inhibited T. gondii growth (50% inhibitory concentration [IC50] = 0.61 μM) with a selective index = 8.48 when tested against hBMECs (50% cytotoxic concentration [CC50] = 5.17 μM). However, IC50 of monensin was 4.13 μM with a SI = 13.82 when tested against microglia cells (CC50 = 57.08 μM), suggesting less sensitivity of microglia cells to monensin treatment. The effect of T. gondii on the integrity of the BBB was assessed by the transendothelial electrical resistance (TEER) assay using an in vitro human BBB model. The results showed that T. gondii infection significantly decreased hBMECs’ TEER resistance, which was rescued when cells were treated with 0.1 µM monensin, probably due to the anti-parasitic activity of monensin. We also investigated the host-targeted effects of 0.1 µM monensin on global gene expression in hBMECs with or without T. gondii infection. Treatment of hBMECs with monensin did not significantly influence the expression of genes involved in the Wnt signalling pathway, suggesting that although inhibition of the Wnt signalling pathway did not play a significant role in T. gondii infection of hBMECs, monensin was still effective in limiting the growth of T. gondii. On the contrary, monensin treatment downregulated pathways related to steroids, cholesterol and protein biosynthesis and their transport between endoplasmic reticulum and Golgi apparatus, and deregulated pathways related to cell cycle and DNA synthesis and repair mechanisms. These results provide new insight into the host-modulatory effect of monensin during T. gondii infection, which merits further investigation.

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