Binding of epithelial cells to lectin-coated surfaces
- 91 Downloads
Epithelial cells may relate to their basement membrane substrates via lectin-like interactions. In a model system for study of this type of interaction, lectin-coated bacteriological plastic petri dishes were presented as substrates for epithelial cell adhesion. Of 21 lectins tested by mixed agglutination against two epithelial cell types, Madin-Darby canine kidney (MDCK), and human embryonic kidney cells (HEK), nine gave less than 5% rosettes and 12 gave 5 to 50% rosettes. Wheat germ agglutinin (WGA) andGeodia cydonium lectin gave the highest percentage of rosettes. Wheat germ agglutinin was readily adsorbed to plastic surfaces and maintained specificity in binding interactions. Both MDCK and HEK cells attached as well to WGA coated petri dishes as to conventional tissue culture dishes. Furthermore, both spread over the lectin-coated surfaces. The MDCK cells grew to confluence and could be subcultured and maintained indefinitely on such surfaces, although WGA in solution was toxic to the cells in concentrations as low as 0.1 to 1.0 µg/ml. Cell attachment to WGA coated dishes was blocked by cycloheximide only if the cells had been preincubated with the inhibitor for several hours. Cell attachment was not inhibited by pretreatment of cells with neuraminidase. Precoating cells with WGA blocked binding to both WGA-coated surfaces and untreated tissue culture dishes. Cells attached to WGA-coated dishes could not be readily dislodged by trypsin-EDTA for the first 2 h after subculture. By 4 h, attachment was again trypsin sensitive, suggesting that the cells synthesized a trypsin-sensitive material that was laid down between the cell surface and the WGA-coated dish. Regeneration of trypsin sensitivity was not blocked by cycloheximide.
Key wordsepithelial cells lectin attachment
Unable to display preview. Download preview PDF.
- 10.Kleinman, H. K.; Hewitt, A. T.; Murray, J. C.; Liotta, L. A.; Rennard, S. I.; Pennypacker, J. P.; McGoodwin, E. B.; Martin, G. R.; Fishman, P. H. Cellular and metabolic specificity in the interaction of adhesion proteins with collagen and with cells. J. Supramol. Struct. 11: 69–78; 1979.PubMedCrossRefGoogle Scholar
- 17.Wu, A. M. T.; Kabat, E. A.; Gruczo, F. G.; Poretz, R. D. Immunochemical studies on the reactivities and combining sites of thed-galacto pyranose and 2-acetamido-2-deoxy-d-galacto pyranose specific lectin purified fromSophra japonica seeds. Arch. Biochem. Biophys. 209: 191–208; 1981.PubMedCrossRefGoogle Scholar
- 19.Kaladas, P. M.; Kabat, E. A.; Kimia, A.; Ersson, B. The specificity of the combining sites of the lectin fromVicia villosa seeds which react with cytotoxic lymphoblasts. Mol. Immunol. In press.Google Scholar
- 20.Wood, C.; Kabat, E. A.; Ebisu, S.; Goldstein, I. J. An immunochemical study of the combining sites of the second lectin isolated fromBandeiraea simplicifolia (BSII). Ann. Immunol. (Inst. Pasteur) 129C: 143–158; 1978.Google Scholar
- 23.Moreno, C.; Kabat, E. A. Studies on human antibodies. VIII. Properties and association constants of human antibodies to blood group A substance purified with insoluble specific adsorbants and fractionally eluted with mono- and oligosaccharide. J. Exp. Med. 129: 871–896; 1969.PubMedCrossRefGoogle Scholar
- 38.Phillips, S. G.; Miller, O. J.; Kabat, E. A. ABH blood group antigens on cultured epithelial cells. Danes, B. S.; Douglas, W. H. J.; Cox, R. P. eds. In vitro epithelia and birth defects. Volume XVI (2) New York: Alan R. Liss, Inc.; 1980; 339–344.Google Scholar