Expression of the intercellular adhesion molecule-1 (ICAM-1) in human renal allografts and cultured human tubular cells.

The in vivo distribution of the intercellular adhesion molecule ICAM-1 was investigated in renal tissue specimens obtained from 17 renal allotransplanted patients, nine normal kidneys (controls), seven native kidneys, nine patients with mesangioproliferative glomerulonephritis, and nine patients with active extracapillary glomerulonephritis. Biopsies from patients with signs of acute rejection showed a significant increase in ICAM-1 expression in the tubular epithelium (P less than 0.05). In normal kidneys (controls) ICAM-1 expression was found in endothelial cells; additional expression in the tubular epithelial cells was induced in patients with extracapillary glomerulonephritis. The in vitro expression of ICAM-1 was examined in cultured human tubular cells after stimulation with gamma-interferon and interleukin-1, treatment with cyclosporin and/or verapamil and coculture with allogenic mononuclear cells. An increased ICAM-1 expression was demonstrated by coculture with allogenic mononuclear cells and after stimulation with gamma-interferon and/or interleukin-1. Cyclosporin or verapamil induced no changes. Our results give support to the hypothesis that ICAM-1 upregulation is important in immune interactions such as allograft rejection. Furthermore, the in vitro model indicates that ICAM-1 expression is regulated by gamma-interferon and interleukin-1 produced by activated T lymphocytes and macrophages.     

慢性排斥移植肾中细胞间粘附分子-1与HLA-DR表达的关系

目的：探讨慢性排斥移植肾中细胞间粘附分子－1（ICAM－1）和HLA－DR表达与间质淋巴细胞浸润的关系。方法：对20例慢性排斥肾移植受者进行肾活检，采用免疫组化技术（ABC法）检测移植肾内ICAM－1和HLA－DR的表达。结果：ICAM－1在慢性排斥移植肾肾小管上皮细胞和间质微小动脉内皮细胞表达增强，而HLA－DR表达则普遍上调，尤其在远曲小管。此外，在ICAM－1和HLA－DR表达增强的局部血管周围和小管间质伴有大量淋巴细胞浸润。结论：慢性排斥移植肾中ICAM－1和HLA－DR表达增强可能在排斥反应中起诱导作用，尤其是间质炎细胞的浸润及抗原递呈，同时它们又可能使表达上调的细胞成为免疫反应效应支的靶细胞，从而参与慢性排斥的细胞免疫损伤及移植肾间质损害过程。     

Probucol inhibits intercellular adhesion molecule-1 expression on cultured rat mesangial cells

SUMMARY: Interleukin-1 (IL-1) has been reported to participate in the progression of glomerulonephritis by, in part, up-regulating intercellular adhesion molecule-1 (ICAM-1) expression in experimental glomerulonephritis. In the present study, we examined whether probucol, an antihyperlipidemic agent, inhibited IL-1-induced inflammatory processes in mesangial cells in culture. Northern blot analysis demonstrated that 200 U/mL IL-1 up-regulated ICAM-1 messenger RNA (mRNA) expression with its peak at 4-6 h after stimulation. Ten μg/mL lipopolysaccharide (LPS), a stimulant to release IL-1 from mesangial cells, induced ICAM-1 mRNA expression by five-fold within 6 h and 10 μg/mL probucol notably reduced this induction. Immunoblotting also confirmed that LPS increased ICAM-1 protein by two-fold within 24 h and probucol inhibited this increase. IL-1 receptor antagonist (IL-1ra; 1–100 ng/mL) suppressed LPS-induced ICAM-1 mRNA expression in a dose-dependent manner and 100 ng/mL IL-1ra completely inhibited ICAM-1 induction, indicating that LPS increased ICAM-1 expression through the action of secreted IL-1. Interleukin-1 activity in culture media, measured by thymocyte proliferation assay, was significantly enhanced by LPS and inhibited by probucol. However, neither LPS nor probucol substantially affected IL-1 mRNA expression, suggesting that the IL-1 activity might be regulated at post-translational level. These results suggest that probucol may act as an anti-inflammatory drug by suppressing IL-1 activity from mesangial cells in the progression of glomerulonephritis.     

Probucol inhibits intercellular adhesion molecule-1 expression on cultured rat mesangial cells

Interleukin-1 (IL-1) has been reported to participate in the progression of glomerulonephritis by, in part, up-regulating intercellular adhesion molecule-1 (ICAM-1) expression in experimental glomerulonephritis. In the present study, we examined whether probucol, an antihyperlipidemic agent, inhibited IL-1-induced inflammatory processes in mesangial cells in culture. Northern blot analysis demonstrated that 200 U/mL IL-1 up-regulated ICAM-1 messenger RNA (mRNA) expression with its peak at 4–6 h after stimulation. Ten μg/mL lipopolysaccharide (LPS), a stimulant to release IL-1 from mesangial cells, induced ICAM-1 mRNA expression by five-fold within 6 h and 10 μg/mL probucol notably reduced this induction. Immunoblotting also confirmed that LPS increased ICAM-1 protein by two-fold within 24 h and probucol inhibited this increase. IL-1 receptor antagonist (IL-1ra; 1–100 ng/mL) suppressed LPS-induced ICAM-1 mRNA expression in a dose-dependent manner and 100 ng/mL IL-1ra completely inhibited ICAM-1 induction, indicating that LPS increased ICAM-1 expression through the action of secreted IL-1. Interleukin-1 activity in culture media, measured by thymocyte proliferation assay, was significantly enhanced by LPS and inhibited by probucol. However, neither LPS nor probucol substantially affected IL-1 mRNA expression, suggesting that the IL-1 activity might be regulated at post-translational level. These results suggest that probucol may act as an anti-inflammatory drug by suppressing IL-1 activity from mesangial cells in the progression of glomerulonephritis.     

Increased expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in rat cardiac allografts

This study was designed to investigate the role of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) during chronic cardiac allograft rejection. Wistar rats were used as donors, and SD rats as recipients heterotopic cardiac transplants. Recipients pretreated with inoculation of donor splenocytes (SPC) followed by cyclophosphamide (CP) were divided into 4 groups: (A) untreated group (n = 18) without immunosuppression; (B) SPC plus CP-treated group (n = 18) that were euthanized at 15-120 days posttransplantation; (C) CsA-treated group (n = 18) euthanized at 2-3 months posttransplantation; and (D) tolerance group (n = 18) treated with SPC plus CP and monitored for at least 1 year posttransplantation. Cardiac allografts were harvested at various times for immunohistochemical studies performed to evaluate the expression of ICAM-1 and VCAM-1. Pretreatment of animals with SPC and CP induced long-term cardiac allograft survival. Immunohistochemical staining demonstrated a low level of ICAM-1 and VCAM-1 expression in cardiac allograft muscle and coronary arteries among Groups B and D. In contrast, the expressions of ICAM-1 and VCAM-1 in cardiac allografts of Groups A and C were significantly higher than those in Groups B and D. Our results suggested that the expression of ICAM-1 and VCAM-1 plays an important role during the development of chronic cardiac allograft rejection.     

细胞间粘附分子-1的反义寡核苷酸在小鼠心脏移植中的作用

目的 探讨细胞间粘附分子-1(ICAM-1)的反义寡核苷酸(ICAM-1-ASO)在小鼠心脏移植中对ICAM-1表达以及在排斥反应中的作用.方法 以BALB/c小鼠为供者,C57小鼠为受者,建立颈部异位心脏移植模型,移植后将受者随机分为3组,每组10只.分别为单纯移植组、对照寡核苷酸组(对照ODN)和ICAM-1-ASO组.各组移植后24 h开始经受者静脉分别注射双蒸水0.3 ml、对照ODN 10 mg/kg和ICAM-1-ASO 10 mg/kg,每天1次,连续5 d.移植后第8天各组随机取5只受者切取移植心,进行病理学检查;采用免疫组织化学方法对移植心组织中ICAM-1的表达进行观察;采用半定量逆转录聚合酶链反应(RT-PCR)对ICAM-1mRNA表达进行定量分析.每组中余下的小鼠继续观察直至移植心停跳,记录各组移植心的存活时间.结果 单纯移植组心脏移植后第8天可观察到明显的排斥反应,移植心心跳减慢、心律不齐、心律紊乱、搏动无力;病理学检查可见大量的淋巴细胞浸润,心肌间质水肿及大片心肌坏死;与移植前对比,内皮细胞和心肌细胞ICAM-1表达明显增强;ICAM-1mRNA表达也明显增加;移植心存活时间平均为(9.8±1.48)d.对照ODN组与单纯移植组差异无统计学意义.而ICAM-1-ASO组与单纯移植组相比,移植心ICAM-1的表达明显减弱,淋巴细胞浸润明显减少,排斥反应程度减轻;移植后第9天心律基本规整,偶有心律不齐,心跳力度较单纯移植组增强;移植心存活时间明显延长至(14.6±1.14)d.结论 ICAM-1-ASO抑制移植心组织中ICAM-1的表达和淋巴细胞的浸润,抑制移植后排斥反应的发生和发展,改善移植物的功能,延长移植物的存活时间,其作用具有序列特异性.     

细胞间粘附分子-1的反义寡核苷酸在异种心脏移植中的作用

目的探讨细胞间粘附分子1的反义寡核苷酸(ICAM1ASO)在小鼠→大鼠的异种心脏移植中对ICAM1表达以及排斥反应的作用。方法BALB/c小鼠和Lewis大鼠分别作为供者和受者,随机配对建立小鼠→大鼠的异种心脏移植模型。按照供者术前处理不同分为双蒸馏水对照组、寡核苷酸对照组和ICAM1ASO组。各组供者术前6h分别从右颈外静脉注射双蒸馏水、寡核苷酸和ICAM1ASO。术后48h各组取5只受者切取供心进行病理学检查;采用免疫组织化学方法对供心组织ICAM1的表达进行观察;半定量逆转录聚合酶链(RTPCR)法对ICAM1mRNA的表达进行定量分析。每组中余下5只受者继续观察直至供心停跳,记录供心存活时间。结果双蒸馏水对照组和寡核苷酸对照组在移植后48h可观察到供心心跳减慢,出现不同程度的心律不齐、心律紊乱,供心搏动无力;供心病理学检查显示心脏组织中广泛间质充血、水肿、出血及血栓形成,部分心肌髓样变性及溶解并伴有炎性细胞浸润;与移植前BALB/c小鼠心脏对比,内皮细胞和心肌细胞ICAM1表达明显增强;ICAM1mRNA表达也明显增加;供心存活时间分别为(53.6±3.58)h和(55.0±3.16)h。ICAM1ASO组供心ICAM1在蛋白水平和mRNA水平的表达均减弱;排斥反应程度也明显减轻;供心存活时间为(66.4±2.61)h;与寡核苷酸对照组和双蒸馏水对照组比较,差异有统计学意义。结论ICAM1ASO能明显抑制异种移植中供心ICAM1的表达,抑制异种移植后排斥反应的发生和发展,延长移植物的存活时间,其作用具有高度序列特异性。     

Clinical significance of intercellular adhesion molecule-1 in ulcerative colitis

This study was conducted to assess the clinical significance of intercellular adhesion molecule-1 (ICAM-1) in ulcerative colitis (UC). The subjects were 53 cases of UC and 43 control cases. ICAM-1 was expressed to a greater degree in the UC specimens. Serum ICAM-1 concentrations in the UC group showed values that were lower than those in the control group (P = 0.0081). Serum ICAM-1 concentrations were found to vary according to degree of clinical severity, activity, and affected range. Considering the cases of surgery, there was only one case in which a lowering of early postoperative values was found. In all cases in which therapeutic drugs including steroids were not administered, late postoperative values were significantly increased. ICAM-1 dynamics in UC were frequently seen in tissue and exhibited a significantly low value in blood serum; however, the influence of clinical severity, activity, diseased area, measuring time, and therapeutic drugs need further study.     

Cloning of porcine intercellular adhesion molecule-1 and characterization of its induction on endothelial cells by cytokines

BACKGROUND: The transplantation of pig organs into humans requires a detailed knowledge of similarities and differences between the two species in the molecular physiology of host defense mechanisms. We therefore set out to identify porcine intercellular adhesion molecule (ICAM)-1 and to characterize its expression by endothelial cells. METHODS: Porcine ICAM-1 cDNA was isolated from an endothelial cell cDNA library. An anti-pig ICAM-1 monoclonal antibody was generated and used to investigate the regulation by cytokines of ICAM-1 expression by porcine aortic endothelial cells (PAEC), using flow cytometry. RESULTS: We found that porcine ICAM-1 was similar in primary structure to human ICAM-1, with five Ig-like domains. COS-7 cells transfected with porcine ICAM-1 supported beta2 but not alpha4 integrin-dependent adhesion of human T lymphoblasts. There was a low-level surface expression of ICAM-1 on unstimulated PAEC and increased expression after stimulation with tumor necrosis factor (TNF)-alpha. However expression of ICAM-1 seemed to be significantly lower than that of vascular cell adhesion molecule-1, both on unstimulated and TNF-alpha-activated PAEC. Recombinant porcine interferon-gamma weakly stimulated ICAM-1 expression when incubated alone with PAEC but had an inhibitory effect on the increase in ICAM-1 due to TNF-alpha, both at 8 and 24 hr. CONCLUSIONS: Our observations confirm the existence of ICAM-1 in the pig and provide novel insights into how porcine and human endothelial cells differ in terms of adhesion molecule expression and cytokine responsiveness. Such differences are potentially important in interpreting models of inflammation in the pig and also in understanding the process of rejection of porcine xenografts.     

Mesangial expression of intercellular adhesion molecule-1 in primary glomerulosclerosis.

Frozen sections of renal biopsy specimens from eight patients with primary focal segmental glomerulosclerosis (FSGS) and 10 patients with membranous nephropathy (MN) were stained in immuno-peroxidase with the intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (MoAb), CL203.4. ICAM-1 was expressed by mesangial cells in six patients with FSGS. On the other hand, ICAM-1 was not detected in mesangial cells in patients with MN or in the non-affected portion of tumoral kidneys used to control normal renal expression of ICAM-1. De novo mesangial expression of ICAM-1 in FSGS suggests that sclerosis results from an inflammatory process, possibly associated with local release of cytokines.     

C-reactive protein induced activation of MAP-K and RANTES in human renal distal tubular epithelial cells in vitro

AIMS: C-reactive protein (CRP) is a component of the acute-phase reaction to inflammation, severe tissue injury, and infection. Investigations have shown that CRP concentration is highly increased in the urine during acute renal graft dysfunction and, therefore, may affect tubular cell metabolism. Nevertheless, no data about the effects of CRP on human renal tubular epithelial cells are available. METHODS: Human renal distal tubular cells (DTC) were isolated immunomagnetically and cultured. Cells were stimulated with affinity chromatography pure native CRP from human ascites (10 - 0.001 microg/ml). Phosphorylation of MAP-K was assessed by Westernblot analysis. Release of RANTES and interleukin-6 was evaluated with an enzyme immunoassay. Cytotoxic effects of CRP were determined by a commercially available Live/Dead assay and MTT assay. Effects on cell proliferation were analyzed by a fluorimetric assay. RESULTS: Westernblot analysis clearly showed that CRP activates the MAP-K pathway of DTC. CRP upregulated RANTES expression of DTC in a significant and dose-dependent manner. CRP (10 microg/ml) induced a 12.3-fold upregulation, CRP 1 or 0.1 microg/ml induced a 6.3-/2.8-fold RANTES upregulation, respectively. Interleukin-6 synthesis was not influenced. Cytotoxic, proliferative or apoptotic effects were not observed at the concentrations used. CONCLUSIONS: We demonstrated an activating effect of CRP on DTC in vitro. In vivo, this effect of CRP might be part of the immune activation cascade during episodes of renal graft rejection or bacterial infections.     

Tubular expression of intercellular adhesion molecule-1 during renal allograft rejection

Molecules responsible for adhesion between cells are known to play an important role in the immune response. The expression of one of these molecules, intercellular adhesion molecule-1 (ICAM-1), was examined on normal and allografted kidneys using a specific monoclonal antibody and an indirect immunoperoxidase technique. The expression of this molecule was compared to that of HLA class II antigens. On normal kidneys and most allograft biopsies taken immediately before implantation, ICAM-1 was expressed only on vascular endothelial cells (VEC) and parietal epithelium of Bowman's capsule. In the 11 kidneys where biopsies were available before and after transplantation, the appearance of rejection was associated with de novo expression of ICAM-1 on renal tubular epithelial cells that closely paralleled that of HLA class II antigens. In addition, an increase in endothelial cell expression of these molecules was also seen in rejection. In 23 random allograft biopsies, most of those with rejection showed tubular expression of both HLA class II antigens and ICAM-1. However, the presence of these molecules on tubules in several biopsies that did not show rejection limits the clinical usefulness of monitoring these antigens in posttransplant biopsies. The upregulation of these molecules is presumed to be secondary to the release of cytokines by cells infiltrating the allograft, although other mechanisms may be operating that explain the expression of these molecules in nonrejecting grafts.     

Transdifferentiation of distal but not proximal tubular epithelial cells from human kidney in culture.

Human renal proximal and distal (thick ascending limb and early distal convoluted tubule) epithelial cells have been isolated according to their specific antigen expression. The cells were well characterized by flow cytometry, enzyme cytochemistry and electron microscopy and cultured for up to 3 months. Cultured tubular cells coexpressed cytokeratin and vimentin as intermediate filament proteins. While primary isolated cells, proximal as well as distal, revealed the phenotypic characteristics of their nephron origin, cultured distal cells showed the tendency to dedifferentiate/transdifferentiate. Distal cells lost their characteristic expression of Tamm-Horsfall glycoprotein and started de novo expression of the proximal marker proteins aminopeptidase M, gamma-glutamyl transferase and dipeptidyl peptidase IV. The expression of these antigens by distal cells could be shown by flow-cytometric analysis and fluorescence microscopy. Enzyme activity assays revealed the activity of aminopeptidase M, gamma-glutamyl transferase and dipeptidyl peptidase IV, but not of the proximal marker enzyme alkaline phosphatase. This antigenic shift could not be prevented in different culture media, and the original phenotype could not be restored. Cultured cells displayed characteristic hormonal stimulation patterns indicative of their proximal and distal origins, as shown by activation of adenylate cyclase by different peptide hormones. These results indicate that distal tubular cells possibly transdifferentiate to a more proximal phenotype in view of loss of the distal marker enzyme Tamm-Horsfall protein and de novo expression of proximal marker enzymes like dipeptidyl peptidase IV and aminopeptidase M.     

Polymerized hemoglobin induces heme oxygenase-1 protein expression and inhibits intercellular adhesion molecule-1 protein expression in human lung microvascular endothelial cells

BACKGROUND: Our clinical trials using a polymerized hemoglobin solution (PolyHb) as a red cell substitute in severely injured patients suggested that this hemoglobin-based oxygen carrier has a systemic antiinflammatory effect. Heme oxygenase-1 (HO-1) has recently been shown to be cytoprotective, and is known to be induced by heme moieties. We investigated the effects of this hemoglobin-based oxygen carrier on HO-1 induction and proinflammatory activation of pulmonary endothelium. STUDY DESIGN: Human lung microvascular endothelial cells were grown to confluence and preincubated with either cell media (control) or with an equal volume mixture of polymerized hemoglobin/cell media (experimental). The cell cultures were subsequently stimulated with lipopolysaccharide. HO-1 expression was detected by protein immunoblot and further quantified by ELISA; intercellular adhesion molecule-1 protein expression was measured by flow cytometry. RESULTS: Polymerized hemoglobin induced synthesis of HO-1 protein in human lung microvascular endothelial cells and, concurrently, inhibited lipopolysaccharide-induced intercellular adhesion molecule-1 protein cell surface expression. CONCLUSIONS: Polymerized hemoglobin attenuates lipopolysaccharide-stimulated expression of intercellular adhesion molecule-1 protein, which is associated with upregulation of the cytoprotective protein HO-1 in human pulmonary endothelial cells. This antiinflammatory effect offers a novel mechanism by which hemoglobin-based oxygen carrier solutions may be exploited therapeutically as resuscitative fluids.