An antiglycolipid antibody inhibits Madin-Darby canine kidney cell adhesion to laminin and interferes with basolateral polarization and tight junction formation.
Epithelial cells polarize not only in response to cell-cell contacts, but also to contacts with a substratum composed of extracellular matrix molecules. To probe the role of specific matrix constituents in epithelial cell polarization, we investigated the effects of an adhesion-blocking mAb, 12B12, on initial polarization of MDCK cells. The 12B12 antibody, raised against whole MDCK cells, blocks adhesion to laminin by 65% but has no effect on adhesion of cells to collagen type I. Taking advantage of this antibody's function-blocking activity, as well as the fact that MDCK cells secrete laminin, the role of endogenous laminin in polarization was examined by plating cells on collagen-coated substrata in the presence of the antibody. Under these conditions, cell spreading was reduced 1.5h after plating, and cells were flatter and had fewer microvilli after 24 h. Even though lateral cell membranes were closely apposed, transepithelial resistance in the presence of the antibody was significantly reduced relative to controls. When the polarization of specific apical and basolateral markers was examined both biochemically and immunocytochemically in the presence of the antibody, we observed that the apical marker polarized at normal rates while basolateral markers did not. Surprisingly, the 12B12 antibody was not directed against any known cell adhesion protein but reacted specifically with Forssman antigen, a glycosphingolipid. These results suggest that glycolipids may play a significant role in cell adhesion via laminin and in epithelial cell polarization.
Zinkl
G M
GM
Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, Massachusetts 02115, USA.
Zuk
A
A
van der Bijl
P
P
van Meer
G
G
Matlin
K S
KS
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001667:Binding, Competitive / Q000276:immunology; D015415:Biomarkers; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D004285:Dogs; D006017:Glycolipids / Q000276:immunology*; D007686:Kidney Tubules, Distal / Q000166:cytology*; D007700:Kinetics; D007797:Laminin / Q000378:metabolism*; D019108:Tight Junctions / Q000502:physiology*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D006017:Glycolipids; D007797:Laminin; D003094:Collagen