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Updated: Aug 15, 2026

Static Adhesion Assay for the Study of Integrin Activation in T Lymphocytes
Published on: June 13, 2014
Integrin alpha 5 beta 1 expression is required for inhibition of keratinocyte migration by ganglioside GT1b
C C Sung1, E A O'Toole, B J Lannutti
1Department of Pediatrics, Northwestern University Medical School, Chicago, Illinois 60614, USA.
Abstract:
Polysialoganglioside GT1b, a keratinocyte membrane glycosphingolipid, inhibits normal keratinocyte adhesion and migration on a fibronectin matrix. The specificity of the inhibition for cells plated on a fibronectin matrix and competition of GT1b inhibition with peptide RGDS suggest that GT1b abrogates the alpha 5 beta 1/fibronectin interaction. We examined the effects of GT1b on the adhesion and migration of keratinocyte-derived cell lines and correlated GT1b responsiveness and alpha 5 beta 1 integrin expression. GT1b (5 nM) significantly inhibited migration of normal human keratinocytes, immortalized keratinocytes, and squamous cell carcinoma SCC12F2 cells on fibronectin, but not on collagen I. Concentrations as high as 5 microM had no effect on SCC13 or HaCaT cells. Likewise, GT1b inhibited fibronectin-dependent cell adhesion of normal human keratinocytes, immortalized keratinocytes, and SCC12F2 cells, but had no effect on SCC13 or HaCaT cells. Flow cytometric and Western immunoblot analysis of integrin expression showed significantly decreased alpha 5 and beta 1 integrin expression in SCC13 and HaCaT cells compared to normal keratinocytes, immortalized keratinocytes, and SCC12F2 cells. Incubation with TGF-beta 1 increased alpha 5 beta 1 integrin expression and induced responsiveness to GT1b in HaCaT cells. These data imply that GT1b "response" requires sufficient expression of alpha 5 beta 1 and further suggest that the mechanism of the inhibitory effect of GT1b involves GT1b/alpha 5 beta 1 interaction.
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