Activation Dependent and Independent VLA-4 Binding Sites on Vascular Cell Adhesion Molecule-1

Needham, Lindsey; Dijk, Sonja Van; Pigott, Rod; Edwards, Richard M.; Shepherd, Maura; Hemingway, Ian; Lindsay, Jack; Clements, John M.

doi:10.3109/15419069409004429

Cited by 38 publications

(31 citation statements)

References 54 publications

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“…The 38-␤1 B lymphoma cell line expressing ␣4␤1 requires a 2-fold greater Mn 2ϩ concentration to achieve half-maximal attachment to CHO monolayers expressing human modules 4 -6 compared with modules 1-3 (19). Adhesion of the U937 human myelomonocytic cell line to module 1 occurs at 37 or 4°C, whereas adhesion to module 4 occurs at 37°C but not 4°C (20). Taken together, these results show that ␣4␤1-mediated adhesion to module 4 requires a different, presumably higher activation state, whereas adhesion to module 1 is constitutive and less dependent on activation.…”

Section: Discussionsupporting

confidence: 55%

Differential Engagement of Modules 1 and 4 of Vascular Cell Adhesion Molecule-1 (CD106) by Integrins α4β1 (CD49d/29) and αMβ2 (CD11b/18) of Eosinophils

Barthel

Annis

Mosher

et al. 2006

Journal of Biological Chemistry

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We have studied adhesion of eosinophils to various forms of vascular cell adhesion molecule 1 (VCAM-1, CD106), an integrin counter-receptor implicated in eosinophil recruitment to the airway in asthma. Full-length 7d-VCAM-1, with seven immunoglobulin-like modules, contains integrin-binding sites in modules 1 and 4. The alternatively spliced six-module protein, 6d-VCAM-1, lacks module 4. In static assays, unactivated purified human blood eosinophils adhered similarly to recombinant soluble human 6d-VCAM-1 and 7d-VCAM-1 coated onto polystyrene microtiter wells. Further experiments, however, revealed differences in recognition of modules 1 and 4. Antibody blocking indicated that eosinophil adhesion to 6d-VCAM-1 or a VCAM-1 construct containing only modules 1-3, 1-3VCAM-1, is mediated by ␣4␤1 (CD49d/29) ,whereas adhesion to a construct containing modules 4 -7, 4 -7VCAM-1, is mediated by both ␣4␤1 and ␣M␤2 (CD11b/18). Inhibitors of phosphoinositide 3-kinase, which block adhesion of eosinophils mediated by ␣M␤2, blocked adhesion to 4 -7VCAM-1 but had no effect on adhesion to 6d-VCAM-1. Consistent with the antibody and pharmacological blocking experiments, eosinophilic leukemic cell lines lacking ␣M␤2 did not adhere to 4 -7VCAM-1 but did adhere to 6d-VCAM-1 or 1-3VCAM-1. Activation of eosinophils by interleukin (IL)-5 enhanced static adhesion to 6d-VCAM-1, 7d-VCAM-1, or 4 -7VCAM-1; IL-5-enhanced adhesion to all 3 constructs was blocked by anti-␣M␤2. Adhesion of unstimulated eosinophils to 7d-VCAM-1 under flow conditions was inhibited by anti-␣4 or anti-␣M. IL-5 treatment decreased eosinophil adhesion to 7d-VCAM-1 under flow, and anti-␣M had the paradoxical effect of increasing adhesion. These results demonstrate that ␣M␤2 modulates ␣4␤1-mediated eosinophil adhesion to VCAM-1 under both static and flow conditions. Vascular cell adhesion molecule 1 (VCAM-1)2 is a multimodular cell-surface glycoprotein up-regulated on human endothelium in response to pro-inflammatory cytokines in the asthmatic lung (1) and has been implicated in recruitment of eosinophils (EOS) from blood to the airway in asthma (2). Heterodimeric integrin receptors interacting with VCAM-1 are important in the rolling, firm adhesion, and movement of EOS (3, 4). These events are regulated by cytokines, which may alter integrin adhesive activity (5, 6). EOS express at least three potential integrin receptors that may be involved in adhesion to VCAM-1: ␣4␤1 (7-9), ␣D␤2 (7), and ␣4␤7 (8 -11).Considerable information is available about features of human VCAM-1 that are important for adhesion. The loop between ␤ strands C and D in the first immunoglobulin (Ig)-like module is accessible on the protein surface and contains the core integrin-recognition sequence Ile 39 -Asp-Ser-Pro-Leu 43 (IDSPL) (11-15). A second IDSPL site is present in the CD loop in module 4 (12, 14). Based on topology of exons within the mammalian genome and the similarity in amino acid sequence, a three-module unit was likely duplicated to generate VCAM-1 modules 1-3 and 4 -6 (16, 17). Modul...

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Section: Discussionsupporting

confidence: 55%

Differential Engagement of Modules 1 and 4 of Vascular Cell Adhesion Molecule-1 (CD106) by Integrins α4β1 (CD49d/29) and αMβ2 (CD11b/18) of Eosinophils

Barthel

Annis

Mosher

et al. 2006

Journal of Biological Chemistry

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“…This suggests that fibronectin, a matrix protein implicated in the pathogenesis of asthma (61)(62)(63), may act as an HBEC-associated ligand for eosinophil ␣ 5 ␤ 1 integrin. In contrast, we could not find a role for ␣ 4 ␤ 1 integrin in adhesion of primed eosinophils to HBEC using a mAb (2B4) that we and others have previously shown reduces eosinophil adhesion to fibronectin and VCAM-1 (35,41,64). Different regions of fibronectin bind to ␣ 5 ␤ 1 and ␣ 4 ␤ 1 integrins; ␣ 5 ␤ 1 binds to the middle portion of the fibronectin polypeptide containing the amino acid sequence Arg-Gly-Asp (RGD), whereas ␣ 4 ␤ 1 binds to a spliced variant containing the connecting segment region (CS-1) (65,66).…”

Section: Discussioncontrasting

confidence: 48%

“…Mouse anti-human ␣ 4 integrin IgG1 (CD49d, clone 2B4) (35) and mouse anti-human ICAM-1 (BBIG-I1) IgG1 whole mAb were gifts from R. Pigott (British Biotech, Oxford, U.K.). F(abЈ) 2 of BBIG-I1 were generated by Cymbus Biotechnology (Chandlers Ford, Hants, U.K.).…”

Section: Antibodiesmentioning

confidence: 99%

TNF-α Potentiates C5a-Stimulated Eosinophil Adhesion to Human Bronchial Epithelial Cells: A Role for α5β1 Integrin

Burke‐Gaffney

Blease²,

Hartnell³

et al. 2002

The Journal of Immunology

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Cooperative action of inflammatory mediators and adhesion molecules orchestrates eosinophil recruitment during allergic inflammation in the airways. This study investigated the mechanisms involved in increasing eosinophil adhesion to human bronchial epithelial cells (HBEC) following priming and activation of eosinophils with TNF-α and complement protein C5a, respectively. Under primed conditions, eosinophil adhesion increased 3-fold from basal (16%), and the effect was significantly greater (p < 0.05) than the increase following stimulation with C5a alone (2-fold). Eosinophil contact with HBEC was essential for priming. In contrast to C5a, adhesion of eotaxin-stimulated eosinophils to HBEC was not primed with TNF-α nor IL-5, a known eosinophil-priming agent. Priming caused activation of αMβ2 integrin; mAb against either the common β2 integrin subunit or its ICAM-1 ligand reduced the primed component of adhesion. Using mAbs against β1 or α5, but not α4 integrin subunit, together with anti-β2 integrin mAb, reduced stimulated adhesion to basal levels. Cross-linking α5β1 integrin increased αMβ2 integrin-dependent adhesion of eosinophils. There are no known adhesion molecule ligands of α5β1 integrin expressed on HBEC; however, fibronectin, the major matrix protein ligand for α5β1 integrin, was detected in association with HBEC monolayers. A mAb against fibronectin, in combination with anti-β2 integrin mAb, reduced adhesion to basal levels. In conclusion, α5β1 integrin may provide a contact-dependent costimulus for eosinophil priming that, together with TNF-α, potentiated C5a activation of αMβ2 integrin and increased eosinophil adhesion to ICAM-1. Fibronectin, associated with HBEC, may act as a ligand for α5β1 integrin. Dual regulation of eosinophil priming may prevent inappropriate activation of eosinophils in the circulation.

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“…By comparing the ability of the two-domain and seven-domain forms to promote melanoma cell spreading, we have shown that the two-domain form has approximately SO% of the celladhesive activity of the seven-domain form. This difference in activity is probably explained by the fact that the sevendomain form contains a second site recognised by a4P1, which is localised in domain 4 (Osborn et al, 1992;Vonderheide and Springer, 1992;Needham et al, 1994). We have recently shown that, in the presence of monoclonal antibodies to either domain 1 or domain 4, the concentration of the seven-domain form required to promote half-maximal spreading is increased approximately two-fold (A. P. Mould, M. J.…”

Section: Discussionmentioning

confidence: 99%

“…The predominant form on cytokine-stimulated endothelium contains seven domains. It has recently been shown that there are two binding sites in VCAM-1 for a4P1, located in domains 1 and 4 (Osborn et al, 1992;Vonderheide and Springer, 1992;Needham et al, 1994), the domain-4 site being absent in the six-domain form. Analysis of the sequence of the sevendomain form reveals 63% sequence identity for the alignment of the regions containing domains 1-3 with that containing domains 4-6, whereas the sequence identity of other pairwise comparisons of the VCAM-1 domains averages 24%.…”

mentioning

confidence: 99%

Expression and Characterization of a Very‐Late Antigen‐4 (α4β1) Integrin‐Binding Fragment of Vascular Cell‐Adhesion Molecule‐1

Dudgeon¹,

Bottomley

Driscoll

et al. 1994

European Journal of Biochemistry

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We have used an Escherichia coli expression system to produce forms of vascular cell-adhesion molecule-1 (VCAM-1) containing the first two and three supposed immunoglobulin-like domains. A form consisting of the first two domains of VCAM-1 is shown to promote very-late antigen-4-dependent spreading of a melanoma cell line comparable to that found for the equivalent region in the full seven-domain form. Preliminary structural analysis by CD and NMR is consistent with an immunoglobulin fold which is predicted from sequence comparison studies.

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Activation Dependent and Independent VLA-4 Binding Sites on Vascular Cell Adhesion Molecule-1

Cited by 38 publications

References 54 publications

Differential Engagement of Modules 1 and 4 of Vascular Cell Adhesion Molecule-1 (CD106) by Integrins α4β1 (CD49d/29) and αMβ2 (CD11b/18) of Eosinophils

Differential Engagement of Modules 1 and 4 of Vascular Cell Adhesion Molecule-1 (CD106) by Integrins α4β1 (CD49d/29) and αMβ2 (CD11b/18) of Eosinophils

TNF-α Potentiates C5a-Stimulated Eosinophil Adhesion to Human Bronchial Epithelial Cells: A Role for α5β1 Integrin

Expression and Characterization of a Very‐Late Antigen‐4 (α4β1) Integrin‐Binding Fragment of Vascular Cell‐Adhesion Molecule‐1

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