VCAM-1 is internalized by a clathrin-related pathway in human endothelial cells but its α4β1 integrin counter-receptor remains associated with the plasma membrane in human T lymphocytes

Ricard, Isabelle; Payet, M; Dupuis, Gilles

doi:10.1002/(sici)1521-4141(199805)28:05<1708::aid-immu1708>3.0.co;2-y

Cited by 43 publications

(26 citation statements)

References 50 publications

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“…MCECs were then incubated at 37°C for an additional 15 minutes to allow time for VCAM-1-mediated internalization (t 1/2 11.2 minutes). 17 Extracellular-restricted phage were removed with 0.2 mol/L glycine, pH 2.2 (3ϫ8 minutes). Internalized phage were recovered by lysis with 0.1% triethanolamine (Sigma) in PBS, pH 7.4, (4 minutes; room temperature).…”

Section: Phage-display Library Selectionmentioning

confidence: 99%

Detection of Vascular Adhesion Molecule-1 Expression Using a Novel Multimodal Nanoparticle

Kelly

Allport

Tsourkas

et al. 2005

Circulation Research

413

360

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Abstract-Endothelial vascular adhesion molecule-1 (VCAM-1) is a critical component of the leukocyte-endothelial adhesion cascade, and its strict temporal and spatial regulation make it an ideal target for imaging and therapy. The goal of this study was to develop novel VCAM-1-targeted imaging agents detectable by MRI and fluorescence imaging using phage display-derived peptide sequences and multimodal nanoparticles (NPs). We hypothesized that VCAM-1-mediated cell internalization of phage display-selected peptides could be harnessed as an amplification strategy to chaperone and trap imaging agents inside VCAM-1-expressing cells, thus improving target-to-background ratios. To accomplish our goal, iterative phage display was performed on murine endothelium under physiological flow conditions to identify a family of VCAM-1-mediated cell-internalizing peptides. One specific sequence, containing the VHSPNKK motif that has homology to the ␣-chain of very late antigen (a known ligand for VCAM-1), was shown to bind VCAM-1 and block leukocyte-endothelial interactions. Compared with VCAM-1 monoclonal antibody, the peptide showed 12-fold higher target-to-background ratios. A VHSPNKK-modified magnetofluorescent NP (VNP) showed high affinity for endothelial cells expressing VCAM-1 but surprisingly low affinity for macrophages. In contrast, a control NP without VCAM-1-targeting sequences showed no affinity for endothelial cells. In vivo, VNP successfully identified VCAM-1-expressing endothelial cells in a murine tumor necrosis factor-␣-induced inflammatory model and colocalized with VCAM-1-expressing cells in atherosclerotic lesions present in cholesterol-fed apolipoprotein E apoE Ϫ/Ϫ mice. These results indicate that: (1) small peptide sequences can significantly alter targeting of NPs, (2) the used amplification strategy of internalization results in high target-to-background ratios, and (3)

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Section: Phage-display Library Selectionmentioning

confidence: 99%

Detection of Vascular Adhesion Molecule-1 Expression Using a Novel Multimodal Nanoparticle

Kelly

Allport

Tsourkas

et al. 2005

Circulation Research

413

360

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“…68,69 There is some evidence that vascular cell adhesion molecule-1 is internalized through clathrin-coated pits. 70 Perhaps adhesion molecules on endothelial cells have evolved to associate with coated pits and other specialized structures to better support the initial capture and rolling of leukocytes under flow. It is noteworthy that both P-selectin and E-selectin must cluster in membrane domains of endothelial cells to enhance their functions, even though they differ significantly in their cell-surface densities and in their affinity and specificity for ligands on leukocytes.…”

Section: E-selectin Clusters In Membrane Domainsmentioning

confidence: 99%

Clustering endothelial E-selectin in clathrin-coated pits and lipid rafts enhances leukocyte adhesion under flow

Setiadi

McEver

2008

Blood

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During inflammation, E-selectin expressed on cytokine-activated endothelial cells mediates leukocyte rolling under flow. E-selectin undergoes endocytosis and may associate with lipid rafts. We asked whether distribution of E-selectin in membrane domains affects its functions. E-selectin was internalized in transfected CHO cells or cytokine-activated human umbilical vein endothelial cells (HUVECs). Confocal microscopy demonstrated colocalization of E-selectin with ␣-adaptin, a clathrin-associated protein.Deleting the cytoplasmic domain of Eselectin or disrupting clathrin-coated pits with hypertonic medium blocked internalization of E-selectin, reduced colocalization of E-selectin with ␣-adaptin, and inhibited E-selectin-mediated neutrophil rolling under flow. Unlike CHO cells, HUVECs expressed a small percentage of E-selectin in lipid rafts. Even fewer neutrophils rolled on E-selectin in HUVECs treated with hypertonic medium and with methyl-␤-cyclodextrin, which disrupts lipid rafts. These data demonstrate that E-selectin clusters in both clathrin-coated pits and lipid rafts of endothelial cells but is internalized in clathrin-coated pits. Distribution in both domains markedly enhances E-selectin's ability to mediate leukocyte rolling under flow. IntroductionRecruitment of leukocytes into secondary lymphoid organs or inflamed tissues requires that the cells first tether to and roll on vascular surfaces. Interactions of selectins with their glycoconjugate ligands mediate tethering and rolling, 1 which precedes integrindependent deceleration, arrest, and transmigration of leukocytes across the endothelial cell layer. 2 L-selectin, expressed on leukocytes, binds to ligands on other leukocytes and on endothelial cells in lymph nodes and in some regions of inflammation. P-and E-selectin, expressed on activated platelets or endothelial cells, bind to ligands on leukocytes. The leukocyte mucin P-selectin glycoprotein ligand-1 (PSGL-1) is the dominant ligand for P-and L-selectin and an important ligand for E-selectin. 3,4 The manner in which selectins or their ligands distribute on cell surfaces has major influence on rolling behavior. 5 For example, L-selectin and PSGL-1 are located on tips of microvilli, 6,7 which enhances the initial tethering of leukocytes. 8 The extended length of P-selectin helps capture flowing neutrophils and slow their rolling velocities, presumably by increasing encounters with PSGL-1. 9 The dimeric structures of P-selectin and PSGL-1 allow them to form dimeric bonds that prolong tether duration and strength. 10 Membrane anchorage of L-selectin and P-selectin through binding of their cytoplasmic domains to cytosolic proteins favors rolling. L-selectin anchors through interactions of its cytoplasmic domain with the cytoskeleton to effectively engage its ligands under flow. 11 P-selectin uses a different mechanism to cluster on the cell surface. Thrombin or histamine mobilizes P-selectin from the membranes of Weibel-Palade bodies to the plasma membranes of endothelial cells. 12,13 Sequences in t...

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“…19 ECV304 cell line, a bladder cancer-derived cell line, 26 has been reported to express many endothelial surface antigens, 27,28 among them is Tie2 (Brizzi et al unpublished data). These cells were transfected with the dominant negative STAT5 (⌬STAT5) construct 13 by the lipofectin methods.…”

Section: Cell Cultures and Transfectionsmentioning

confidence: 99%

Angiopoietin 2 Induces Cell Cycle Arrest in Endothelial Cells: A Possible Mechanism Involved in Advanced Plaque Neovascularization

Calvi

Dentelli

Pagano

et al. 2004

ATVB

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Objective-To characterize the molecules and the mechanisms regulating the neoangiogenetic process in advanced atherosclerotic plaques. Methods and Results-Western blot and immunofluorescence analysis of atherosclerotic specimens demonstrated that unlike neovessels from early lesions that expressed vascular endothelial growth factor (VEGF) and angiopoietin1 (Angio1), vessels from advanced lesions expressed VEGF and angiopoietin 2 (Angio2). Moreover, only few neovessels from advanced lesions showed a positive immunostaining for proliferating cell nuclear antigen. Angio1-elicited and Angio2-elicited intracellular events in endothelial cells (EC) demonstrated that while Angio1 triggered Erk1/Erk2 mitogen activated protein kinases (MAPK) and Akt activation, Angio2 (50 ng/mL) induced STAT5 activation and p21 waf expression and increased the fraction of cells in G1. Both Angio2-mediated events were abrogated by expressing a dominant negative STAT5 construct (⌬STAT5). Consistent with the expression of Angio2 in neovessels of advanced lesions a transcriptionally active STAT5 was detected. Moreover, co-immunoprecipitation experiments revealed the presence of a STAT5/Tie2 molecular complex in neointima vessels from advanced, but not from early, lesions. 3 Consistently, in vivo developing primary lesions might be considered as underperfused and dependent on neovessel formation for continued growth. 4 Angiogenesis in mature tissues is composed of a series of cellular responses including activation of EC proliferation by angiogenic factors, such as VEGF, fibroblast growth factor (FGF), and angiopoietins (Angio) and formation of tube-like structures and vessel stabilization. 5 Compared with the developmental angiogenesis, the role of angiopoietins in pathological angiogenesis is less understood. The angiopoietin family consists of four members, Angio1 to 4. 6 -8 Angio1 and 4 stimulate their receptor tyrosine kinase Tie2, whereas Angio2 and 3 by binding to Tie2 antagonize Angio1 signaling. 6 -8 Signal transduction pathways via Tie2 and VEGF receptors (VEGFR) have been extensively examined. 9,10 In particular, Korpelainem et al showed that Tie2 and VEGFR1 overexpression triggers the phosphorylation of the signal transducer and activator of transcription (STAT) 3 and STAT5. 11 STAT5 belongs to the family of transcriptional factors, which, on activation, binds to the receptor via their src homology 2 domain (SH2). 12 Recruited STATs, dimerize, translocate to the nucleus, and activate transcription of target genes mainly involved in regulation of cell cycle progression. 12,13 Cell cycle phases are coordinated by regulatory proteins denoted as cyclins and cyclin-dependent kinases (CDKs). 14 Phase-specific cyclin/CDK complexes confer specificity and orderly progression through the cell cycle. Initially, cyclin D/CDK4 or cyclin E/CDK2 complexes, in cooperation with proliferating cell nuclear antigen (PCNA), coordinate DNA duplication by regulating the transition through the G1 and S phase. 15 Furthermore, cell cycle progression is ...

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VCAM-1 is internalized by a clathrin-related pathway in human endothelial cells but its α4β1 integrin counter-receptor remains associated with the plasma membrane in human T lymphocytes

Cited by 43 publications

References 50 publications

Detection of Vascular Adhesion Molecule-1 Expression Using a Novel Multimodal Nanoparticle

Detection of Vascular Adhesion Molecule-1 Expression Using a Novel Multimodal Nanoparticle

Clustering endothelial E-selectin in clathrin-coated pits and lipid rafts enhances leukocyte adhesion under flow

Angiopoietin 2 Induces Cell Cycle Arrest in Endothelial Cells: A Possible Mechanism Involved in Advanced Plaque Neovascularization

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