A study of the structure, function and distribution of beta 5 integrins using novel anti-beta 5 monoclonal antibodies.
Here, we have utilized six new anti-human beta 5 monoclonal antibodies to perform a detailed investigation of the structure, function and distribution of beta 5 integrins. Monoclonal anti-beta 5 specificity was confirmed by reactivity with beta 5-transfected CHO cells, by direct binding to the beta 5 subunit (immunoblotting), and by immunodepletion experiments using polyclonal anti-beta 5 serum. The beta 5 subunit was predominantly associated with the alpha v subunit, although on some cell lines, the level of beta 5 exceeded that of alpha v for unknown reasons. Cell adhesion studies showed that the adhesive function of beta 5 could be stimulated, inhibited or unaltered by different anti-beta 5 monoclonal antibodies. The beta 5 subunit was involved in adhesion to both vitronectin and fibronectin and, at least for K562 cells, fibronectin appeared to be the preferred ligand. Flow-cytometry studies showed that the beta 5 subunit was expressed at moderate to high levels on all adherent cell lines examined, was absent from all lymphoid cell lines, and was only weakly expressed on myeloid cell lines. Staining of thymic sections showed the distribution of beta 5 on blood vessels, Hassal's corpuscles, cortical and medullary stromal cells, and basement membranes. Skin sections showed beta 5 on the basal layer of the epidermis and on some dermal blood vessel walls, and kidney sections showed staining of glomerular regions, juxta glomerular apparatus, proximal convoluted tubules and collecting tubules, and at least one anti-beta 5 antibody also stained epithelial cells of proximal tubules.
Pasqualini
R
R
Division of Tumor Virology, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA 02115.
Bodorova
J
J
Ye
S
S
Hemler
M E
ME
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D016466:CHO Cells; D002448:Cell Adhesion / Q000187:drug effects; D006224:Cricetinae; D005353:Fibronectins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000737:chemistry* / Q000378:metabolism; D007668:Kidney / Q000737:chemistry; D012867:Skin / Q000737:chemistry; D013329:Structure-Activity Relationship; D013950:Thymus Gland / Q000737:chemistry; D014407:Tumor Cells, Cultured; D019096:Vitronectin
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D006023:Glycoproteins; D016023:Integrins; D019096:Vitronectin