Vascular Cell Adhesion Molecule-1 Staining Research Articles

Relationship between vascular cell adhesion molecule-1 and morbidly adherent placenta

IntroductionMorbidly adherent placenta is a clinical condition of which prevalance is steadily increasing. It is described as the invasion of the placenta into the uterine wall through the myometrium and beyond. Vascular cell adhesion molecule-1 is a calcium independent transmembrane glycoprotein. Several studies have shown that vascular cell adhesion molecule-1 increases invasion in tumor cells. In our study we investigated the expression of vascular cell adhesion molecule-1 which has been shown to increase invasion in various tumors in morbidly adherent placenta cases. Material and methodThis is a prospective case–control study. Eighty-four patients who were diagnosed with morbidly adherent placenta and 96 patients without any risk factors as a control group were included in the study. Vascular cell adhesion molecule-1 staining was examined by immune-histochemical staining in placental samples. ResultsThe patients included in the study were similar in terms of demographic characteristics. Of the 84 patients in the morbidly adherent placenta group, 65 (74.7%) showed positive staining. This number was 39 (40.6%) in the control group. Staining in morbidly adherent placenta group was statistically significantly higher (p<0.05). DiscussionThe etiology of morbidly adherent placenta is still a clinically unknown condition. This study is the first study on this field in the literature. In our study, we showed that vascular cell adhesion molecule-1 expression increased in morbidly adherent placenta group. Although the entire pathogenesis is not explained this can be considered as a step for understanding this subject.

A Sativex(®) -like combination of phytocannabinoids as a disease-modifying therapy in a viral model of multiple sclerosis.

Sativex(®) is an oromucosal spray, containing equivalent amounts of Δ(9) -tetrahydrocannabinol (Δ(9) -THC) and cannabidiol (CBD)-botanical drug substance (BDS), which has been approved for the treatment of spasticity and pain associated to multiple sclerosis (MS). In this study, we investigated whether Sativex may also serve as a disease-modifying agent in the Theiler's murine encephalomyelitis virus-induced demyelinating disease model of MS. A Sativex-like combination of phytocannabinoids and each phytocannabinoid alone were administered to mice once they had established MS-like symptoms. Motor activity and the putative targets of these cannabinoids were assessed to evaluate therapeutic efficacy. The accumulation of chondroitin sulfate proteoglycans (CSPGs) and astrogliosis were assessed in the spinal cord and the effect of Sativex on CSPGs production was evaluated in astrocyte cultures. Sativex improved motor activity - reduced CNS infiltrates, microglial activity, axonal damage - and restored myelin morphology. Similarly, we found weaker vascular cell adhesion molecule-1 staining and IL-1β gene expression but an up-regulation of arginase-1. The astrogliosis and accumulation of CSPGs in the spinal cord in vehicle-infected animals were decreased by Sativex, as was the synthesis and release of CSPGs by astrocytes in culture. We found that CBD-BDS alone alleviated motor deterioration to a similar extent as Sativex, acting through PPARγ receptors whereas Δ(9) -THC-BDS produced weaker effects, acting through CB2 and primarily CB1 receptors. The data support the therapeutic potential of Sativex to slow MS progression and its relevance in CNS repair.

Periadventitial atRA citrate-based polyester membranes reduce neointimal hyperplasia and restenosis after carotid injury in rats.

Oral all-trans retinoic acid (atRA) has been shown to reduce the formation of neointimal hyperplasia; however, the dose required was 30 times the chemotherapeutic dose, which already has reported side effects. As neointimal formation is a localized process, new approaches to localized delivery are required. This study assessed whether atRA within a citrate-based polyester, poly(1,8 octanediolcitrate) (POC), perivascular membrane would prevent neointimal hyperplasia following arterial injury. atRA-POC membranes were prepared and characterized for atRA release via high-performance liquid chromatography with mass spectrometry detection. Rat adventitial fibroblasts (AF) and vascular smooth muscle cells (VSMC) were exposed to various concentrations of atRA; proliferation, apoptosis, and necrosis were assessed in vitro. The rat carotid artery balloon injury model was used to evaluate the impact of the atRA-POC membranes on neointimal formation, cell proliferation, apoptosis, macrophage infiltration, and vascular cell adhesion molecule 1 (VCAM-1) expression in vivo. atRA-POC membranes released 12 μg of atRA over 2 wk, with 92% of the release occurring in the first week. At 24 h, atRA (200 μmol/l) inhibited [(3)H]-thymidine incorporation into AF and VSMC by 78% and 72%, respectively (*P = 0.001), with negligible apoptosis or necrosis. Histomorphometry analysis showed that atRA-POC membranes inhibited neointimal formation after balloon injury, with a 56%, 57%, and 50% decrease in the intimal area, intima-to-media area ratio, and percent stenosis, respectively (P = 0.001). atRA-POC membranes had no appreciable effect on apoptosis or proliferation at 2 wk. Regarding biocompatibility, we found a 76% decrease in macrophage infiltration in the intima layer (P < 0.003) in animals treated with atRA-POC membranes, with a coinciding 53% reduction in VCAM-1 staining (P < 0.001). In conclusion, perivascular delivery of atRA inhibited neointimal formation and restenosis. These data suggest that atRA-POC membranes may be suitable as localized therapy to inhibit neointimal hyperplasia following open cardiovascular procedures.

Positive C4d staining of the portal vein endothelium in the liver of patients with biliary atresia: a role of humoral immunity in ongoing liver fibrosis.

This study aimed to clarify the role of complement activation in fibrogenesis in BA. In total, 27 paraffin-embedded liver biopsy samples were immunohistochemically analyzed using C4d polyclonal antibody, vascular cell adhesion molecule-1 (VCAM-1), and CD45. The liver samples were obtained from 25 patients during Kasai operation, and two additional specimens were obtained from 2 patients by needle biopsy later at the time of liver function deterioration. The degree of liver fibrosis was histologically graded 1-3. Among the 25 samples, 9 showed C4d-positive immunostaining localized on the endothelia of a few portal veins in the portal tract. The degree of fibrosis was correlated with C4d staining (p=0.025). The age at Kasai operation correlated with the degree of fibrosis and the C4d positivity. Two needle biopsy samples were positive for C4d. Among 13 samples submitted for VCAM-1 staining, 2 negative samples were C4d negative and all positive C4d samples were VCAM-1 positive with CD45 mononuclear cell infiltration. These findings suggest that ongoing cirrhosis could be a result of progressive "vasculopathy" of the portal vein caused by humoral and cell-mediated immune interaction.

Abstract 845: Vascular cell adhesion molecule-1 (VCAM-1) expression in patients with ovarian tumors of low malignant potential

Abstract The molecular basis for gynecologic tumor dissemination into the peritoneum is not well understood, however it is the primary mechanism of death in advanced stage ovarian cancers. Although serous borderline and serous invasive ovarian cancers are similar, they differ in their ability to implant and invade peritoneal structures. Vascular cell adhesion molecule-1 (VCAM-1), a cell surface receptor expressed on activated endothelial and mesothelial cells, regulates leukocyte attachment and extravasation at sites of inflammation. Recent studies by our group demonstrated the ability of ovarian carcinoma to induce the expression of VCAM-1 on mesothelial cells, and correlated the presence of VCAM-1 with peritoneal dissemination of the tumor. We undertook the current study to determine whether VCAM-1 is expressed on the mesothelium of patients with borderline ovarian tumors. After IRB approval was obtained, cell blocks containing peritoneum and omentum from 10 patients with borderline tumors were processed and stained using antibodies to VCAM-1. Mesothelial expression of VCAM-1 was determined by a single pathologist blinded to tumor type and stage. Samples were determined to be positive if mesothelial cells demonstrated membrane staining. Cytoplasmic staining for VCAM-1 was not considered positive, nor was staining of non-mesothelial cells. Of the 10 patients, four were stage 1a and six were stage 1c serous borderline tumors. Nine of 10 had peritoneal or omental biopsies that did not stain for VCAM-1. The one sample which did stain positively was micropapillary type, which is an invasive sub-type of borderline ovarian cancers. Together, these observations support the hypothesis that VCAM-1 expression correlates with ovarian cancer peritoneal invasion and offers a possible explanation for the inability of borderline tumors to invade peritoneal structures. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 845.

Renal vascular inflammation induced by Western diet in ApoE-null mice quantified by 19F NMR of VCAM-1 targeted nanobeacons

We have designed multifunctional nanoparticulate reporter bioprobes capable of targeting vascular cell adhesion molecule 1 (VCAM-1), which is up-regulated in numerous inflammatory processes. These perfluorocarbon-cored nanoparticles emit a unique 19F magnetic resonance (MR) signature, providing the potential to localize and quantify VCAM-1 expression in early atherosclerosis. Nanoparticle-VCAM-1 targeting specificity was confirmed by in vitro binding and competition studies. ApoE-null and control C57-BL6 mice (n = 6/group), fed a Western diet for 35 weeks, were injected i.v. with targeted or non-targeted nanoparticles. After two hours, kidneys were excised and prepared for analysis. ApoE-null kidneys exhibited increased VCAM-1-targeted nanoparticle content over healthy controls by 19F MR spectroscopy (36.5 +8.8 vs. 9.3 +2.2 × 10 8/g, P < .05), which correlated with increased VCAM-1 staining (2.5 ± 1.3% vs. 0.9 ± 0.3%, P < .05); their relative biodistributions were confirmed by fluorescence microscopy and MR imaging. These molecular imaging agents offer new approaches for detection, quantification, and longitudinal evaluation of early inflammation utilising 19F MR spectroscopy and imaging. From the Clinical Editor Multifunctional nanoparticulate reporter bioprobes capable of targeting vascular cell adhesion molecule 1 (VCAM-1) are reported in this paper. These perfluorocarbon-cored nanoparticles offer new approaches for detection, quantification, and longitudinal evaluation of early inflammation utilising 19F MR spectroscopy and imaging.

Activation of the blood–brain barrier by SIV (simian immunodeficiency virus) requires cell-associated virus and is not restricted to endothelial cell activation

The primary cell infected during acute HIV neuropathogenesis is the monocyte-derived macrophage. We have demonstrated that there is activation of the BBB (blood-brain barrier) during acute viral infection and at terminal AIDS. However, it has never been determined if there is a requirement for the virus to be carried through the BBB or how these Trojan horses would be induced to cross the BBB. We added SIVmac251-infected (SIV is simian immunodeficiency virus) mononuclear cells (and their supernatants) to the luminal or abluminal compartment of our BBB model. There was activation of both sides of the BBB model, only if viral-infected cells were added to the luminal compartment, as opposed to the addition of cell-free supernatants. This suggests that cell-associated virus is essential for the activation of the BBB. This, in turn, would be expected to lead to further infiltration of cells capable of viral infection. VCAM-1 (vascular cell adhesion molecule 1) staining revealed, for the first time, that there is an absolute requirement for virally infected cells, and not just the presence of virus for the activation of the BBB.

Expression of adhesion molecules by cultured spiral ligament fibrocytes stimulated with proinflammatory cytokines.

Secondary cultures from murine spiral ligament (SL) fibrocytes were stimulated with proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), and expression of various adhesion molecules was investigated. Cultures without cytokine stimulation did not show positive immunostaining for vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), or mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Although staining was also negative after stimulation with IL-1beta, VCAM-1 and ICAM-1 staining was observed after the cells were stimulated with TNF-alpha. Reverse transcription-polymerase chain reaction analysis showed messenger RNAs for both VCAM-1 and ICAM-1 expression to be present after fibrocytes were stimulated with TNF-alpha. These data suggest that activated fibrocytes may cause inflammatory cells to persist in the SL. Given that SL fibrocytes may play a role in homeostasis of cochlear fluid and ion concentrations, prolongation of the inflammatory response could lead to fibrocyte damage that might ultimately result in cochlear malfunction.

Co-Infection with Chlamydia pneumoniae and Helicobacter pylori Results in Vascular Endothelial Dysfunction and Enhanced VCAM-1 Expression in ApoE-Knockout Mice

Background: Upregulation of proinflammatory endothelial cell adhesion molecules and decreased bioactivity of endothelial nitric oxide (NO) are important in the pathogenesis of atherosclerosis. We investigated the effects of co-infection with Chlamydia pneumoniae and Helicobacter pylori on these two events in apoE-KO mice. Methods: Thirty-two apoE-KO mice, 8 weeks old, were equally divided into 4 groups. The first 2 groups were infected with either C. pneumoniae or H. pylori, while the 3rd group was infected with both C. pneumoniae and H. pylori. Mice from the 4th group and 4 wild-type mice served as controls. Thoracic and abdominal aortas were harvested after 10 weeks, and staining for vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 was analyzed by immunocytochemistry. The endothelial vasomotor responses of thoracic aortas to methacholine were studied in organ chambers in the absence and presence of L-NAME. The plasma levels of nitrate/nitrite were measured. Results: Staining for VCAM-1 was more intense at the branching sites of aortas from mice with co-infection than in mono-infected or noninfected apoE-KO mice. The relaxation responses to methacholine and the plasma levels of nitrate/nitrite were significantly less in the co-infected group than in the other groups (p < 0.05). Conclusion: Co-infection of apoE-KO mice with C. pneumoniae and H. pylori seems to be associated with impaired bioactivity of endothelial NO and increased expression of VCAM-1 at branching sites. The findings may suggest an additive interaction of these pathogens in atherogenesis.

Inducible nitric oxide synthase deficiency does not affect the susceptibility of mice to atherosclerosis but increases collagen content in lesions.

Although endothelial nitric oxide synthase (NOS) is antiatherogenic, the role of inducible NOS (iNOS) in the development of atherosclerosis is not established. We compared the susceptibility of iNOS knockout (iNOS(-/-)) and wild-type (iNOS(+/+)) mice to the development of atherosclerosis induced by feeding an atherogenic diet for 15 weeks. Plasma lipid level, atherosclerotic lesion size, and cellular density in the lesions were all similar in the 2 strains (lesion size: iNOS(+/+) 285+/-73x10(3) microm(2), iNOS(-/-) 293+/-82x10(3) microm(2), n=10). iNOS mRNA was detected in the lesions of iNOS(+/+) but not iNOS(-/-) mice through RT-PCR. Immunohistochemically, iNOS(+/+) mice showed iNOS staining in macrophages and medial smooth muscle cells in the lesions. Nitrotyrosine staining showed a similar distribution, whereas it was absent in iNOS(-/-) mice. There was no apparent difference in the intensity or distribution of vascular cell adhesion molecule-1 staining in the lesions of the 2 strains. However, the lesions of iNOS(+/+) mice showed a markedly decreased extracellular collagen content compared with those of iNOS(-/-) mice iNOS induction does not affect the development of atherosclerosis in mice fed an atherogenic diet, but the resulting lesions show decreased levels of extracellular collagen and may be more fragile.

Expression of Adhesion Molecules in Kidney with Experimental Chronic Obstructive Uropathy: The Pathogenic Role of ICAM-1 and VCAM-1

Background: Chronic obstructive uropathy induced by maintained unilateral ureter ligation in the rat is characterized morphologically by interstitial inflammation, interstitial fibrosis, and tubular atrophy. Infiltrating mononuclear inflammatory cells, particularly T lymphocytes and macrophages, may contribute to the progression of this lesion by mediating tubular injury and by the activation of interstitial fibroblasts, with resultant tubular atrophy and interstitial fibrosis, respectively. Altered expression and activation of adhesion molecules by leukocytes, vascular endothelial cells, and parenchymal cells likely contributes both to the infiltration of inflammatory cells into the tubulointerstitial compartment and to the interaction of activated inflammatory cells with parenchymal cells. Methods: In the current study, we examined changes in the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in a 90-day model of maintained unilateral ureter ligation in male Sprague-Dawley rats. Results: Rat kidneys showed constitutive expression of ICAM-1 mRNA and constitutive immunostaining for ICAM-1 in peritubular capillaries, glomeruli, and a small percentage of cortical tubules. Ureter ligation resulted in a rapid increase in ICAM-1 mRNA, which was almost 2-fold greater than those of the contralateral and control kidneys as early as 3 h and which was maintained at a 4- to 6-fold higher level in the ligated vs. contralateral kidneys throughout the entire 90-day time course. There was a marked increase in ICAM-1 immunostaining within the tubular epithelium, with up to 80% of both cortical and medullary tubular cross-sections showing strong apical immunostaining from day 6 to 25, with a subsequent decrease throughout the remainder of the experiment. ICAM-1 immunostaining in the expanding interstitium in the ligated kidneys showed a gradual increase throughout the duration of the experiment. In contrast, glomerular immunostaining for ICAM-1 was decreased in the ligated compared to the contralateral kidneys throughout the entire experiment. There was a later but prominent increase in VCAM-1 mRNA in ligated kidneys, which was first evident at 2 days and which was maintained 2- to 10-fold greater than the contralateral kidneys throughout the entire time course. VCAM-1 immunostaining increased in the expanding interstitium, but decreased in glomeruli in obstructed vs. contralateral kidneys. Tubular staining for VCAM-1 did not change after ureter ligation. Conclusion: Increased ICAM-1 and VCAM-1 may contribute to the prominent inflammatory cell infiltration in the chronic tubulointerstitial nephritis accompanying maintained unilateral ligation. Tubule expression of ICAM-1, which occurs during a similar time course as previously documented for tubular cell proliferation and especially tubular cell apoptosis in this model, may contribute to injurious interactions of activated inflammatory cells with tubular epithelium.

Expression of inducible cell adhesion molecules in the normal human lung: immunohistochemical study of their distribution in pulmonary blood vessels

The distribution of cell adhesion molecules in the normal human lung was investigated using antibodies to E-selectin, P-selectin, intercellular adhesion molecule 1(ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1). Lectin staining by Ulex europaeus type I agglutinin (UEA I) and immunohistochemistry for von Willebrand factor (vWF) was used to visualize a maximum of blood vessels per section. In the bronchial mucosa, staining for P-selectin was positive in ca 90%, and staining for E-selectin, VCAM-1, and ICAM-1 was positive in 40-70% of the vessels stained with UEA I. In the pulmonary circulation (vasa publica) ca 90% of non-capillary vessels stained by anti-vWF expressed P-selectin, 54% VCAM-1, 41% E-selectin, and only ca 20% ICAM 1. The alveolar capillaries were stained consistently by UEA I, but not by the panel of antibodies tested. The alveolar epithelium and, inconstantly, basal cells of the bronchial epithelium were positive for ICAM-1. The distribution pattern of inducible adhesion molecules in normal human lung tissue suggests that a permanent low-grade endothelial activation may exist in particular in the mucosa of the airways, which could be due to the normal antigen exposure via inhaled air.

Upregulation of VCAM-1 and ICAM-1 at atherosclerosis-prone sites on the endothelium in the ApoE-deficient mouse.

Focal recruitment of monocytes and lymphocytes is one of the earliest detectable cellular responses in the formation of lesions of atherosclerosis. This localized accumulation of leukocytes is a multistep process in which the endothelium remains intact and may regulate leukocyte recruitment by expressing specific adhesion molecules. To examine the relationship of adhesion molecule expression to initiation factors and the sites of lesion formation, we analyzed the expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and platelet-endothelial cell adhesion molecule-1 (PECAM-1) en face on the aortic endothelium of control mice and homozygous apolipoprotein E-deficient (ApoE -/-) mice that develop complex lesions of atherosclerosis similar to those in humans. In control mice, VCAM-1 staining was weak and limited to sites of altered blood flow. In contrast, in the ApoE -/- mice, VCAM-1 appeared to be localized over the surface of groups of endothelial cells in lesion-prone sites. Expression of VCAM-1 preceded lesion formation, and increased expression above control levels appeared to be correlated with the extent of exposure to plasma cholesterol. Although ICAM-1 was the most prominent adhesion molecule in lesion-prone sites, its expression appeared to be independent of plasma cholesterol levels and was upregulated in both ApoE -/- and control mice. At lesion-prone sites associated with altered blood flow, ICAM-1 was located over the surface of each endothelial cell and on microvilli, whereas VCAM-1 was confined to the cell periphery in non-lesion-prone sites. PECAM-1 was localized at the cell periphery throughout the aorta, and its expression did not appear to be regulated. Thus, the levels, localization, and characteristics of expression of VCAM-1, ICAM-1, and PECAM-1 appear to be differentially regulated. Upregulation of VCAM-1 and ICAM-1 is associated with sites of lesion formation.

Role of Adhesion Molecule Expression and Soluble Fractions in Hepatic Resection

Background: Little has so far been documented about the relationship between liver injury and adhesion molecules. The aim of this study is to clarify the role of adhesion molecules in hepatic resection by studying both the expression of such adhesion molecules and the measurement of their soluble fractions in the blood.Study Design: To study adhesion molecule expression in the liver, liver biopsies were obtained before and after hepatectomy in 14 patients. Using frozen sections, immunochemical staining for intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was then performed. To study the soluble fractions of adhesion molecules in the hepatic venous blood, the serum soluble fractions of ICAM-1 and VCAM-1 from another 17 patients were measured using an enzyme-linked immunosorbent assay. The plasma levels of polymorphonuclear leukocyte (PMN) elastase were also measured using an enzyme immunoassay. Both the preoperative and postoperative values of the serum soluble fractions of ICAM-1, VCAM-1, and PMN elastase were then compared. The correlation between their values and the perioperative variables was also investigated.Results: Either ICAM-1 or VCAM-1 was stained on the sinusoidal endothelial cells and Kupffer cells or circulating PMNs in the sinusoid. The positive rate of either ICAM-1 or VCAM-1 staining in livers with more than 40 minutes of total ischemic time (80%) was significantly higher than that in livers with less than 40 minutes of total ischemic time (0%; p < 0.05). The incidence of postoperative complications in the ICAM-1 positive staining group tended to be higher than that in the ICAM-1 negative group.Both soluble fractions of ICAM-1 and VCAM-1 in patients with cirrhotic liver disease were also significantly higher than those in patients with a normal liver. The soluble VCAM-1 level in patients with a chronic active hepatitic liver tended to be higher than that in those with a nonactive hepatitic liver. The preoperative level of soluble ICAM-1 correlated with that of VCAM-1, PMN elastase, albumin, aspartate aminotransferease (AST), and the indocyanine green dye retention test at 15 minutes (ICG R15), while the preoperative level of VCAM-1 correlated with albumin, the hepaplastin test, AST, and ICG R15. Both the serum soluble ICAM-1 and VCAM-1 levels after hepatectomy were significantly lower than those before hepatectomy. By contrast, the posthepatectomy level of PMN elastase was significantly higher than its prehepatectomy level. The difference between the postoperative and preoperative values of soluble ICAM-1 correlated with the postoperative AST level, postoperative alanine aminotransferase level, and total ischemic time.Conclusions: Adhesion molecules were expressed in the liver after hepatic resection, and such expression correlated with a total ischemic time during hepatectomy. In addition, judging from the soluble forms of such molecules, these adhesion molecules play an important role in hepatic resection.

Detection of the chemokine RANTES and endothelial adhesion molecules in nasal polyps

Background: To better understand the mechanisms of eosinophil recruitment into the upper airways, we examined human nasal polyps for the expression of the chemotactic cytokine RANTES and the endothelial adhesion molecules E-selectin and vascular cell adhesion molecule-1 (VCAM-1). Methods: Routine histologic examination and immunostaining with antibodies to RANTES, E-selectin, and VCAM-1 were performed on three types of tissues: nasal polyps, sinus mucosa, or turbinates from patients undergoing other elective procedures (S/T), and nasal biopsy specimens from nonallergic volunteers (NA). To further quantify the expression of endothelial adhesion molecules, some tissue samples were homogenized, and the resulting supernatants were assayed with sandwich ELISAs for VCAM-1 and E-selectin. Results: Polyp eosinophil counts ranged from 19/mm 2to 1818/mm 2(763 ± 120/mm 2 mean ± SEM) and were significantly higher than those found in the control tissues (5 ± 2 in S/T samples and 20 ± 9 in NA samples, p< 0.002). Immunochemical staining for RANTES was observed in 11 of 14 polyps; intense staining for RANTES (grade 3) was observed in six of 14 polyps. None of nine S/T samples or five NA samples demonstrated grade 3 staining. Staining with anti-RANTES was largely localized to airway and glandular epithelium. There was no significant correlation between counts of eosinophils or the combined total of eosinophils plus mononuclear cells and the intensity of epithelial RANTES staining in all nasal tissues. Staining for VCAM-1, as well as for E- selectin, was detected in 11 of 14 polyps and eight of 13 control tissues. VCAM-1 detected by ELISA in polyp tissues (6.8 ± 1.3 μ g/gm) was higher than that found in six S/T samples (1.2 ± 0.3 μ g/gm, p < 0.005) and in two NA samples (1.8 ± 0.02 μ g/gm, p = 0.08). E-selectin values in polyps (1.4 ± 0.3 μ g/gm) were not statistically different from those detected in six S/T samples (0.5 ± 0.2 μ g/gm) or two NA samples (1.6 ± 0.4 μ g/gm). Counts of eosinophils and eosinophils plus mononuclear cells displayed a strong correlation with VCAM-1 ELISA values ( p < 0.005 and p lt; 0.004, respectively) but not with VCAM-1 staining. VCAM-1 staining correlated with EG2-positive eosinophils in nasal polyp tissues ( p < 0.01). E-selectin staining did not correlate with either neutrophil or eosinophil counts. Conclusions: These studies demonstrate that the chemokine RANTES is produced in vivo and is localized predominantly to nasal epithelium. Endothelial activation, as indicated by adhesion molecule expression, occurs in human nasal polyp tissues and in control tissues, possibly reflecting the continued antigen exposure of the nasal mucosa. The correlations found in this study suggest that expression of VCAM-1 plays a role in the selective recruitment of eosinophils and mononuclear cells into nasal polyp tissues and that RANTES may be more important in localizing eosinophils to the epithelium. (J A LLERGY C LIN I MMUNOL 1996;98:766-80.)

Expression of VCAM-1 in rabbit iliac arteries is associated with vasodilator dysfunction of regenerated endothelium following balloon injury

Previous studies have demonstrated impaired endothelium-dependent vasodilation following balloon injury of the rabbit iliac artery, suggesting dysfunction of the regenerated endothelium. More recently, expression of vascular cell adhesion molecule-1 (VCAM-1) has been shown up to 4 weeks in a different injury model of the rabbit aorta, suggesting sustained inflammatory activation of endothelium following injury. The aim of the present study was to combine the examination of VCAM-1 expression, as a marker of cellular activation, and the assessment of endothelium-dependent relaxation to test the hypothesis that different forms of altered endothelial function are concurrently present in the chronic phase following experimental balloon angioplasty. New Zealand White rabbits fed either a standard ( n = 7) or a 1% cholesterol ( n = 8) diet, underwent balloon injury of the iliac artery 5 weeks following the initiation of the diet. Four weeks after balloon injury, control and balloon-injured arteries were harvested for in vitro studies of vascular reactivity, for morphometric analysis and for immunocytochemical staining with Rb 1 9 monoclonal antibody directed against VCAM-1 and with CD 31 monoclonal antibody for the identification of endothelial cells. The combination of balloon injury and hypercholesterolemia resulted in a marked impairment of endothelium-dependent relaxation to acetylcholine and in a pronounced intimal proliferation compared to control or either intervention alone. Control rings of rabbits fed a normal diet did not reveal positive staining for VCAM-1. Balloon-injured rings of the animals fed a normal diet showed focal areas of positive staining in the superficial cell layer overlying intimal lesions. In the group fed a high cholesterol diet, control rings and ballooned rings showed positive staining for VCAM-1 in cells overlying intimal lesions. In all groups the superficial cell layers were identified as endothelial cells by positive staining for CD 31. In conclusion, the present study shows that regenerated endothelium following mechanical arterial injury reveals expression of VCAM-1 together with impaired receptor-mediated vasodilator capacity. Thus, the expression of VCAM-1 and the impairment of endothelium-dependent relaxation may represent different features of endothelial dysfunction following balloon injury which may actively influence the proliferative lesion of restenosis after balloon angioplasty.

Immunolocalization of adhesion molecules in psoriatic arthritis, psoriatic and normal skin

Adhesion molecule expression in synovial membrane obtained from patients with psoriatic arthritis (PA) has previously been compared with rheumatoid arthritis (RA). Although expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was similar in both psoriatic and rheumatoid synovium, in contrast, little or no endothelial leucocyte adhesion molecule-1 (ELAM-1) was observed in psoriatic synovium. In the present study, the expression of ICAM-1, ELAM-1 and VCAM-1 was examined in the involved and uninvolved skin from patients with PA (n = 15), patients with psoriasis (Ps) but no arthritis (n = 5) and in normal skin (n = 4). ICAM-1 was intensely expressed on endothelium and keratinocytes of involved skin from patients with Ps with or without arthritis. There was constitutive expression of ICAM-1 on endothelium only in uninvolved and normal skin. In contrast, ELAM-1 expression was restricted to endothelial cells; it was widespread and intense in involved skin, but was minimal in uninvolved and normal skin. VCAM-1 was expressed on endothelium, and also on some dendritic cells in involved psoriatic skin. There was minimal VCAM-1 staining on endothelial cells in uninvolved and normal skin. In conclusion, in involved psoriatic skin from patients with and without arthritis ICAM-1, ELAM-1 and VCAM-1 expression is up-regulated on vascular endothelium, and ICAM-1 is expressed on keratinocytes. However, ELAM-1 and VCAM-1 expression seen in dermal vessels is not found in psoriatic synovial vessels. These differences suggest a mechanism for controlling cellular traffic in Ps and in PA.

Cell Adhesion Molecule Expression within the Microvasculature of Human Colorectal Malignancies

In situ expression of intercellular adhesion molecule-1 (ICAM-1), endothelial leukocyte adhesion molecule-1 (ELAM-1), and vascular cell adhesion molecule-1 (VCAM-1) was investigated in 20 human colorectal cancers using immunohistochemical techniques. Tumor microvessels, detected with endothelial marker Ulex Europaeus Agglutinin-I (UEAI), were consistently present within both stromal and glandular areas. Vessels expressing cell adhesion molecules (CAMs) were less frequent but more common within the tumor stroma. Although ICAM-1 and ELAM-1 vessel staining was present in all tumors, ICAM-1 staining was the most consistent with primary localization to stroma while ELAM-1 was variable with localization to both stromal and glandular areas. VCAM-1 staining was inconsistent and was rare in glandular areas. A significant increase in the number of vessels expressing CAMs with a concomitant decrease in the total number of vessels was noted in bowel muscularis adjacent to tumor compared to remote bowel. No relationship between number of vessels or frequency of CAM positive vessels and tumor site, grade, or stage was noted. These studies demonstrate enhanced microvascular expression of CAMs in close proximity to colorectal tumors despite decreases in total number of vessels, suggesting that factors within the tumor microenvironment effect tumor microvascular development. Correlation between these studies and previous microscopic studies suggest that vessels expressing CAMs play a role in immune cell infiltration and may provide new targets for anti-tumor therapies.

Estradiol-17β regulates the induction of VCAM-1 mRNA expression by interleukin-1β in human umbilical vein endothelial cells

We examined the effect of estradiol-17β on the expression of vascular cell adhesion molecule-1 (VCAM-1), an adhesion molecule, in human umbilical vascular endothelial cells. After preincubation with estradiol-17β for 24 hours, cells were treated for 4 h with 0.5 μg/ml recombinant human interleukin-1β. The RNase protection assay was performed using an [α- 32P]-labeled 121 base pair VCAM-1 cRNA probe. Preincubation with estradiol-17β (250 or 500 pg/ml) suppressed the induction of VCAM-1 mRNA expression by interleukin-1β. VCAM-1 staining with a monoclonal antibody decreased when cells were incubated with estradiol-17β at 250 and 500 pg/ml, while staining was detectable when cells were treated with interleukin-1β at 0.5 μg/ml. In conclusion, estradiol-17β regulates the induction of VCAM-1 gene expression by interleukin-1β in human umbilical vein endothelial cells.

Expression of endothelial and leukocyte adhesion molecules intercellular adhesion molecule-1, E-selectin, and vascular cell adhesion molecule-1 in the bronchial mucosa in steady-state and allergen-induced asthma

Background: Interactions between cell adhesion molecules and their ligands are an integral part of inflammatory processes and may have direct relevance to the pathology of asthma. Methods: Immunostaining with antibodies to cell adhesion molecules was performed on bronchial biopsy specimens from persons with intrinsic and extrinsic asthma, normal nonasthmatic control subjects, and patients with asthma after allergen challenge. Results: There was constitutive expression of intercellular adhesion molecule-1 (ICAM-1), E-selectin and vascular cell adhesion molecule-1 (VCAM-1) in patients with intrinsic and extrinsic asthma and in control subjects. Compared with control subjects, ICAM-1 E-selectin staining in the submucosa was greater in the intrinsic asthmatic group for intensity ( p < 0.02, p < 0.05) and extent ( p < 0.01, p < 0.05) of staining, respectively. No differences were observed between patients with extrinsic asthma and normal control subjects, and VCAM-1 expression did not differ among the groups. Epithelial expression of ICAM-1 was more frequent in patients with asthma compared with normal control subjects ( p < 0.05). Compared with diluent challenge, bronchial biopsy specimens obtained 24 hours after allergen challenge revealed no significant differences in intensity or extent of staining for ICAM-1, E-selectin, or VCAM-1. After allergen challenge, the intensity and extent of both VCAM-1 and ICAM-1 expression correlated significantly with the number of eosinophils (cells positive for major basic protein). Epithelial ICAM-1 expression was more frequently observed after allergen challenge than after diluent challenge ( p < 0.02). Conclusions: The data suggest a complex pattern of regulation for ICAM-1, E-selectin, and VCAM-1 in vivo, where they may reflect the degree of ongoing inflammation in asthma.