Expression Of Including Intercellular Adhesion Molecule-1 Research Articles

PbO nanoparticles increase the expression of ICAM-1 and VCAM-1 by increasing reactive oxygen species production in choroid plexus.

PbO nanoparticles (nano-PbO) are widely used in the production of electrode materials, but exposure to them can cause brain damage. The first barrier preventing nano-PbO from entering the brain is the choroid plexus. However, the effect of nano-PbO on the choroid plexus remains unclear. Thus, the purpose of this study was to investigate the effect of nano-PbO exposure on lymphocyte cells infiltration, the adhesion protein of the choroid plexus as well as the role of reactive oxygen species (ROS) during the process. Results showed that nano-PbO exposure increased the percentage of lymphocyte cells in the brain and upregulated the expression of surface adhesion proteins, including intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in choroid plexus. Meanwhile, nano-PbO treatment also resulted in the increase of intercellular ROS production, and significantly decrease glutathione (GSH) content, glutathione peroxidase (GSH-PX) activity, and superoxide dismutase (SOD) activity in Z310 cells beside the increase of ICAM and VCAM-1 expression. Treatment with ROS inhibitor N-acetylcysteine (NAC) significantly downregulated the expression of ICAM-1 and VCAM-1expression. In conclusion, exposure to nano-PbO increases the expression of ICAM-1 and VCAM-1 through oxidative stress, which may contribute to peripheral lymphocyte cells infiltration into the brain.

Abstract 12416: Modeling Diabetic Endothelial Cell Dysfunction With Induced Pluripotent Stem Cells

Introduction: Diabetes is one of the major risk factors for cardiovascular diseases (CVDs), which is mediated by vascular endothelial dysfunction. Currently, there is no in vitro disease model of endothelial cell (EC) dysfunction in diabetic patients, which hinders mechanistic understanding and therapeutic discovery. The goal of this study is to develop a comprehensive model of diabetic EC dysfunction using patient-derived induced pluripotent stem cells (iPSCs) and recapitulating the diabetic microenvironment. Methods: Our model utilizes iPSC derived ECs from the following sources: healthy subjects and diabetic patients with underlying CVDs (n=3). To simulate in situ conditions of diabetes, we used the following treatment for 5 days: hyperglycemia (25 mM glucose); high urea (9 mM); and inflammatory stimulation with tumor necrosis factor alpha (TNF-α, 8 pg/ml). EC dysfunction was assessed through oxidative stress, inflammation, permeability, and thrombosis. Immunofluorescence identified localization for surface adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1), endothelial selectin (E-sel), and platelet selectin (P-sel). Enzyme-linked immunosorbent assay (ELISA) quantitatively measured expression of these surface adhesion molecules. Results: Overall, the qualitative effect of the diabetic microenvironment resulted in increased expression of ICAM-1, E-sel, and P-sel. After TNF-α treatment, we observed a stronger increase in expression of P-sel in healthy subject-derived cells than diabetic patient-derived cells. This indicates a possible pre-conditioned tolerance of pro-inflammatory conditions. Ongoing experimentation will provide results for quantitative expression of ICAM-1, E-sel, and P-sel after treatment with the diabetic microenvironment. Conclusion: The development of a robust in vitro model of diabetic endothelial dysfunction, similar to the onset of type 2 diabetes, will allow for high-throughput testing of pharmacological agents to ultimately treat the disease’s effects within the vascular bed. The widespread implications of type 2 diabetes on the vascular bed further amplify the significance of this study in the development of pharmacological agents to treat the disease.

C1q/TNF-related protein-9 inhibits cytokine-induced vascular inflammation and leukocyte adhesiveness via AMP-activated protein kinase activation in endothelial cells

Although recent studies have reported cardioprotective effects of C1q/TNF-related protein 9 (CTRP9), the closet adiponectin paralog, its role on cytokine-induced endothelial inflammation is unknown. We investigated whether CTRP9 prevented inflammatory cytokine-induced nuclear factor-kappa B (NF-κB) activation and inhibited the expression of adhesion molecules and a chemokine in the vascular endothelial cell. We used human aortic endothelial cells (HAECs) to examine the effects of CTRP9 on NF-κB activation and the expression of NF-κB-mediated genes, including intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and monocyte chemoattractant protein-1 (MCP-1). Tumor necrosis factor alpha (TNFα) was used as a representative proinflammatory cytokine. In an adhesion assay using THP-1 cells, CTRP9 reduced TNFα-induced adhesion of monocytes to HAECs. Treatment with CTRP9 significantly decreased TNFα-induced activation of NF-κB, as well as the expression of ICAM-1, VCAM-1, and MCP-1. In addition, treatment with CTRP9 significantly increased the phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), the downstream target of AMPK. The inhibitory effect of CTRP9 on the expression of ICAM-1, VCAM-1, and MCP-1 and monocyte adhesion to HAECs was abolished after transfection with an AMPKα1-specific siRNA. Our study is the first to demonstrate that CTRP9 attenuates cytokine-induced vascular inflammation in endothelial cells mediated by AMPK activation.

Plasmin induces intercellular adhesion molecule 1 expression in human endothelial cells via nuclear factor-κB/mitogen-activated protein kinases-dependent pathways

Activation of endothelial cells (ECs) by proinflammatory stimuli triggers expression of cellular adhesion molecules including intercellular adhesion molecule 1 (ICAM-1) on the cell surface. Such molecules mediate the transendothelial migration of inflammatory cells, which is an early key step of atherogenesis. We have previously demonstrated that plasmin activates human inflammatory cells via the annexin A2 heterotetramer (A2t). Here we show that human umbilical vein endothelial cells (HUVECs) and human microvascular endothelial cells express high amounts of A2t, as shown by Western blotting, fluorescence microscopy and flow cytometry. Activation of HUVEC by plasmin led to cleavage of the annexin A2 subunit of the receptor complex, followed by the activation of Akt/nuclear factor (NF)-κB signaling, and phosphorylation of MAP kinases p38 and ERK1/2. Further, plasmin stimulates the NF-κB/p38-dependent expression of ICAM-1 by HUVEC. The plasmin-induced activation of cells was abolished when annexin A2 was down-regulated by small-interfering RNA. In vivo, we show co-localization of the ECs marker CD31 with the plasmin receptor A2t and ICAM-1 in human atherosclerotic plaques of human femoral arteries, which also exhibit activated NF-κB signaling as revealed by immunofluorescence staining for phosphorylated IκBα. In addition, plasma of patients with advanced atherosclerosis exhibited enhanced plasmin activity and up-regulated levels of plasmin-α2-antiplasmin. These data point to a previously unrecognized functional role of plasmin in EC biology, which could be of particular relevance in the development of atherosclerosis.

Rhein Inhibits the Expression of Vascular Cell Adhesion Molecule 1 in Human Umbilical Vein Endothelial Cells with or without Lipopolysaccharide Stimulation

Reducing the expression of endothelial cell adhesion molecules (ECAMs) is known to decrease inflammation-induced vascular complications. In this study, we explored whether rhein can reduce the inflammation-induced expression of ECAMs in human umbilical vein endothelial cells (HUVECs) with or without lipopolysaccharide (LPS) stimulation. HUVECs were treated with different concentrations of rhein with or without 2.5 μg/ml LPS stimulation. Cell viability was assayed using the MTT method. Real-time PCR and Western blot analysis were used to measure the transcription and expression levels of ECAMs, including intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-SELECTIN and related signaling proteins. The results indicated that rhein (0-20 μmol/L) and LPS (0-10 μg/ml) had no effect on the viability of HUVECs. LPS could promote the expression of VCAM-1, ICAM-1 and E-SELECTIN. Rhein appeared to target VCAM-1, ICAM-1 and E-SELECTIN, with the transcription and expression of all three factors being reduced by the rhein treatment (10 and 20 μmol/L). The transcription and expression of VCAM-1 were also reduced by treatment with rhein (10 and 20 μmol/L) in the presence of LPS stimulation. In conclusion, rhein treatment reduced the expression of VCAM-1 in HUVECs via a p38-dependent pathway.

Antifibrotic effects of tetrandrine on hepatic stellate cells and rats with liver fibrosis

Anti-inflammation strategies are one of the proposed therapeutic approaches to hepatic fibrosis. Tetrandrine (C(38)H(42)O(8)N(2), molecular weight: 622; Tet), an alkaloid isolated from the Chinese medicinal herb Stephania tetrandra, has been shown to exert anti-inflammatory activity in pulmonary diseases. The purpose of the present study was to investigate the in vitro and in vivo effects of Tet on hepatic fibrosis. A cell line of rat hepatic stellate cells (HSC-T6) was stimulated with transforming growth factor-beta1 (TGF-beta1) or tumor necrosis factor-alpha (TNF-alpha). The inhibitory effects of Tet on the nuclear factor kappaB (NFkappaB) signaling cascade and molecular markers including intercellular adhesion molecule-1 (ICAM-1) and alpha-smooth muscle actin (alpha-SMA) secretion were assessed. Fibrosis was induced by dimethylnitrosamine (DMN) administration in rats for 4 weeks. Fibrotic rats were randomly assigned to one of the four groups: vehicle (0.7% carboxyl methyl cellulose, CMC), Tet (1 mg/kg), Tet (5 mg/kg), or silymarin (50 mg/kg), each given by gavage twice daily for 3 weeks starting after 1 week of DMN administration. At the end of the study, liver tissues were scored for fibrosis and analyzed for molecular markers of fibrosis. Tetrandrine (0.5-5.0 micromol/L) concentration-dependently inhibited NFkappaB transcriptional activity induced by TNF-alpha, including IkappaBalpha phosphorylation and mRNA expressions of ICAM-1 in HSC-T6 cells. In addition, Tet also inhibited TGF-beta1-induced alpha-SMA secretion and collagen deposition in HSC-T6 cells. Fibrosis scores of livers from DMN-treated rats with high-dose Tet (1.3 +/- 0.3) were significantly reduced in comparison with DMN-treated rats receiving saline (2.0 +/- 0.2). Hepatic collagen content of DMN rats was significantly reduced by either Tet or silymarin treatment. Double-staining results showed that alpha-SMA- and NFkappaB-positive cells were decreased in the fibrotic livers by Tet and silymarin treatment. In addition, mRNA expression of ICAM-1, alpha-SMA, and TGF-beta1 was attenuated by Tet treatment. Moreover, levels of plasma aspartate aminotransferase and alanine aminotransferase activities were reduced by Tet and silymarin treatment. Tetrandrine exerts antifibrotic effects in both HSC-T6 cells and in rats with DMN-induced fibrosis.

Ethanol inhibits prolactin- and tumor necrosis factor-?-, but not gamma interferon-induced expression of intercellular adhesion molecule-1 in human astrocytoma cells

In humans, alcohol consumption has multiple effects on the immune system. Despite an increase in our understanding of the effects of alcohol on the immune system, little is known about the effect of alcohol on the neuroimmune response. In the central nervous system (CNS), astrocytes and microglial function as immune effector cells. In response to infection of injury, astrocytes increase in number and size, express several proinflammatory cytokines, MHC class I and II antigens, and several adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1). Interactions between ICAM-1 and its counter-receptors play an important role in the regulation of neuroimmune response. In this study, cultured human astrocytoma cells were used to examine the effect of ethanol on ICAM-1 expression. Western blot analyses show that quiescent astrocytes express, at least, four immunoreactive ICAM-1 proteins with apparent molecular weights 55, 67, 82, and 90 kDa. Incubation of human astrocytoma cells with tumor necrosis factor-alpha (TNF-alpha) or prolactin (PRL) resulted in marked increases in all four immunoreactive ICAM-1 proteins. In the presence of ethanol, however, PRL- and TNF-alpha-induced increases in all four immunoreactive ICAM-1 proteins were markedly inhibited. ICAM-1 is a cell surface transmembrane glycoprotein. Using a cell surface specific ICAM-1 adhesion assay we found that in human astrocytoma cells TNF-alpha, interferongamma (IFN-gamma) and PRL increased cell surface ICAM-1 expression. Consistent with our Western blot analyses, ethanol significantly inhibited TNF-alpha- and PRL-induced cell surface ICAM-1 expression. By contrast, IFN-gamma-induced ICAM-1 expression was not inhibited by exposure of the cells to ethanol. Expression of ICAM-1 is regulated predominantly at the transcriptional level. In the present report, we show that TNF-alpha increased ICAM-1 mRNA levels in human astrocytoma cells and that ethanol markedly blocked TNF-alpha-induced increases in ICAM-1 mRNA levels. Further, we found that PRL-induced ICAM-1 expression was, at least in part, due to a PRL-induced increase in TNF-alpha syntheses and secretion. Our results clearly indicate that ethanol has a pronounced effect on ICAM-1 expression in human astrocytoma cells, thus suggesting that ETOH exposure may impair the immune response in the CNS by blocking leukocytes adhesion and migration into the CNS in response to injury or infection.

Ethanol inhibits prolactin- and tumor necrosis factor-α-, but not gamma interferon-induced expression of intercellular adhesion molecule-1 in human astrocytoma cellsPresented in part at the 79th Annual Meeting of the Endocrine Society, Minneapolis, Minnesota.

In humans, alcohol consumption has multiple effects on the immune system. Despite an increase in our understanding of the effects of alcohol on the immune system, little is known about the effect of alcohol on the neuroimmune response. In the central nervous system (CNS), astrocytes and microglial function as immune effector cells. In response to infection of injury, astrocytes increase in number and size, express several proinflammatory cytokines, MHC class I and II antigens, and several adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1). Interactions between ICAM-1 and its counter-receptors play an important role in the regulation of neuroimmune response. In this study, cultured human astrocytoma cells were used to examine the effect of ethanol on ICAM-1 expression. Western blot analyses show that quiescent astrocytes express, at least, four immunoreactive ICAM-1 proteins with apparent molecular weights 55, 67, 82, and 90 kDa. Incubation of human astrocytoma cells with tumor necrosis factor-α (TNF-α) or prolactin (PRL) resulted in marked increases in all four immunoreactive ICAM-1 proteins. In the presence of ethanol, however, PRL- and TNF-α-induced increases in all four immunoreactive ICAM-1 proteins were markedly inhibited. ICAM-1 is a cell surface transmembrane glycoprotein. Using a cell surface specific ICAM-1 adhesion assay we found that in human astrocytoma cells TNF-α, interferonγ (IFN-γ) and PRL increased cell surface ICAM-1 expression. Consistent with our Western blot analyses, ethanol significantly inhibited TNF-α- and PRL-induced cell surface ICAM-1 expression. By contrast, IFN-γ-induced ICAM-1 expression was not inhibited by exposure of the cells to ethanol. Expression of ICAM-1 is regulated predominantly at the transcriptional level. In the present report, we show that TNF-α increased ICAM-1 mRNA levels in human astrocytoma cells and that ethanol markedly blocked TNF-α-induced increases in ICAM-1 mRNA levels. Further, we found that PRL-induced ICAM-1 expression was, at least in part, due to a PRL-induced increase in TNF-α syntheses and secretion. Our results clearly indicate that ethanol has a pronounced effect on ICAM-1 expression in human astrocytoma cells, thus suggesting that ETOH exposure may impair the immune response in the CNS by blocking leukocytes adhesion and migration into the CNS in response to injury or infection. J. Cell. Biochem. 77:455–464, 2000. © 2000 Wiley-Liss, Inc.

Virulent Treponema pallidum 47 kDa antigen regulates the expression of cell adhesion molecules and binding of T-lymphocytes to cultured human dermal microvascular endothelial cells.

Perivasculitis and endothelial cell abnormalities are prominent histopathologic features of syphilis. Various cutaneous lesions are the main clinical features of syphilis. We examined whether Treponema pallidum 47 kDa antigen regulates the expression of cell adhesion molecules on human dermal microvascular endothelial cells (HDMEC) and the regulation of T-lymphocytes binding to HDMEC. Using immunofluorescence flow cytometry and enzyme-linked immunosorbent assay (ELISA), we demonstrated that T. pallidum upregulated the expression of adhesion molecules, including intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E-selectin. The 47 kDa antigen of T. pallidum also activated endothelium as measured by the upregulation of the expression of adhesion molecules on HDMEC, and it also promoted an increased adherence of T-lymphocytes to HDMEC. The expressions of ICAM-1 and VCAM-1 on HDMEC and the adherence of T-lymphocytes to HDMEC were inhibited by treatment with anti-TNF-alpha antibody or anti-IL-1alpha antibody. These results show that T. pallidum or T. pallidum-specific 47 kDa antigen are capable of stimulating HDMEC to increase the expression of ICAM-1, VCAM-1 and E-selectin and thereby, promote the adherence of T-lymphocytes. The whole process may play an important role in the immunopathogenesis of syphilis and it is likely that TNF-alpha and IL-1alpha are involved.

Mast cells of psoriatic and atopic dermatitis skin are positive for TNF-alpha and their degranulation is associated with expression of ICAM-1 in the epidermis.

The release of cytokines from cutaneous cells may be of major importance in the initiation and development of many inflammatory skin disorders. For example, tumor necrosis factor-alpha (TNF-alpha), which in healthy skin is found preformed only in mast cells, is able to induce the expression of several adhesion molecules including intercellular adhesion molecule-1 (ICAM-1). Increased expression of ICAM-1 occurs in keratinocytes in lesional skin of psoriasis and atopic dermatitis (AD) and it is considered to be an important initiator of leucocyte/keratinocyte interactions in skin inflammation. We counted the mast cells showing TNF-alpha immunoreactivity using a double-staining method in nonlesional and lesional skin sections from 12 patients with AD and 12 patients with psoriasis. The percentage of TNF-alpha+ mast cells in lesional and nonlesional AD skin was 36 +/- 22% and 21 +/- 15% (P < 0.018, paired t-test), respectively, and in psoriatic skin was 16 +/- 25% and 15 +/- 15%, respectively (P < 0.89, paired t-test). We also cultured whole skin biopsies taken from the healthy-looking skin of psoriatic and AD patients in the presence of mast cell degranulator compound 48/80, which resulted in focal expression of ICAM-1 in the epidermis. In cultured keratinocytes, both histamine and an extract of a human mast-cell line (HMC-1) induced ICAM-1 immunostaining only in occasional cells, but the combination of histamine and the HMC-1 extract resulted in intense ICAM-1 staining in numerous cells. This enhancement of ICAM-1 staining was abolished by preincubation of the HMC-1 extract with anti-TNF-alpha antibody. These results suggest that the degranulation of mast cells induces the expression of ICAM-1 in keratinocytes probably via TNF-alpha and histamine.

Heparin Regulates ICAM-1 Expression in Human Endothelial Cells: An Example of Non-Cytokine-Mediated Endothelial Activation

Activated endothelial cells up-regulate the expression of several molecules on their plasma membranes, including intercellular adhesion molecule-1 (ICAM-1). The role of heparin in regulating endothelial cell gene expression is unclear. We thus have investigated the ability of heparin to regulate ICAM- gene expression by using flow cytometry and the ribonuclease protection assay with human umbilical vein and aortic endothelial cells cultured in growth medium supplemented with 90 [microg/ml heparin (heparin-sufficient, HS) or in growth medium without added heparin (heparin-deficient, HD). We found that HD medium increased plasma membrane protein and mRNA for ICAM-1 but not for HLA-DR, even though both ICAM-1 and HLA-DR protein and mRNA were inducible by gamma interferon (IFN-gamma). In addition, phorbol ester and IFN-gamma increased the expression of plasma membrane ICAM-1 or ICAM-1 and HLA-DR, respectively, more in HD medium than in HS medium. We found that the HD-mediated increase of ICAM- mRNA was reversible by the addition of heparin, and that the half-life of ICAM-1 mRNA was the same in both HS- and HD-treated cells. Also, heparin was found to suppress increases in ICAM-1 mRNA at a concentration as low as 5 microg/ml. These findings indicate that heparin deficiency induces endothelial activation characterized by increased ICAM-1, and that such induction is not dependent on cytokines or endotoxin. The modulation of ICAM-1 expression by heparin appears to occur at the transcriptional level. Thus, heparin may have a role in regulating endothelial function by affecting the expression of ICAM-1, thereby impacting upon the trans-endothelial trafficking of leukocytes.

Modulation of the antigenic phenotype of human melanoma cells by differentiation-inducing and growth-suppressing agents.

Tumor cells often display alterations in their normal program of cellular differentiation. A promising approach for the treatment of cancer involves the induction of terminal differentiation and a loss of proliferative capacity in cancer cells. In human melanoma cells, the combination of mezerein (MEZ) and fibroblast interferon (IFN-beta), results in a rapid and irreversible suppression of cell growth with a concomitant increase in the synthesis of melanin. The induction of terminal differentiation is associated with alterations in the expression of several cellular genes, including fibronectin, ISG-15 and ISG-54, and changes in the expression of specific cell surface antigens, including intercellular adhesion molecule-1 (ICAM-1) and HLA Class I antigens. In the HO-1 human melanoma cell line, induction of terminal differentiation by MEZ plus IFN-beta results in an induction and/or increased expression of ICAM-1, HLA Class I antigens and HLA Class II antigens. IFN-beta and MEZ alone can modulate expression of these antigens to a lower extent than does the combination of compounds. Induction of terminal differentiation and the irreversible suppression of cell growth is not a prerequisite for antigenic modulation in HO-1 cells. This is indicated by the inability of immune interferon (IFN-gamma), a strong inducer of ICAM-1, HLA Class I antigens and HLA Class II antigens synthesis, or the combination of IFN-beta plus IFN-gamma which synergistically but reversibly suppresses HO-1 growth, to induce melanin synthesis or terminal differentiation in HO-1 cells.(ABSTRACT TRUNCATED AT 250 WORDS)