Differential engagement of modules 1 and 4 of vascular cell adhesion molecule-1 (CD106) by integrins alpha4beta1 (CD49d/29) and alphaMbeta2 (CD11b/18) of eosinophils.
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 alpha4beta1 (CD49d/29), whereas adhesion to a construct containing modules 4-7, 4-7VCAM-1, is mediated by bothalpha4beta1 andalphaMbeta2 (CD11b/18). Inhibitors of phosphoinositide 3-kinase, which block adhesion of eosinophils mediated by alphaMbeta2, 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 alphaMbeta2 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-alphaMbeta2. Adhesion of unstimulated eosinophils to 7d-VCAM-1 under flow conditions was inhibited by anti-alpha4 or anti-alphaM. IL-5 treatment decreased eosinophil adhesion to 7d-VCAM-1 under flow, and anti-alphaM had the paradoxical effect of increasing adhesion. These results demonstrate that alphaMbeta2 modulatesalpha4beta1-mediated eosinophil adhesion to VCAM-1 under both static and flow conditions.
Barthel
Steven R
SR
Departments of Biomolecular Chemistry and Medicine, University of Wisconsin, 4285A Medical Sciences Center, 1300 University Avenue, Madison, WI 53706-1532, USA.
Annis
Douglas S
DS
Mosher
Deane F
DF
Johansson
Mats W
MW
D001665:Binding Sites; D039481:CD11b Antigen / Q000378:metabolism*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D004804:Eosinophils / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D006801:Humans; D039041:Integrin alpha4beta1 / Q000378:metabolism*; D019169:Jurkat Cells; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D039481:CD11b Antigen; D039041:Integrin alpha4beta1; D011994:Recombinant Proteins; D019010:Vascular Cell Adhesion Molecule-1