Studies of Lewis antigens and H. pylori adhesion in CHO cell lines engineered to express Lewis b determinants

Glycobiology
Jonas LöflingJan Holgersson

Abstract

Many microbes bind and adhere via adhesins to host cell carbohydrates as an initial step for infection. Therefore, cell lines expressing Lewis b (Le(b)) determinants were generated as a potential model system for Helicobacter pylori colonization and infection, and their expression of blood group Lewis determinants was characterized. CHO-K1 cells were stably transfected with selected glycosyltransferase cDNAs, and two Le(b) positive clones, 1C5 and 2C2, were identified. Expression of Lewis (Le(a), Le(b), Le(x), and Le(y)) determinants was analyzed by flow cytometry of intact cells, SDS-PAGE/Western blot of solubilized glycoproteins, and thin layer chromatography immunostaining of isolated glycolipids (GL). Binding of H. pylori to cells was examined by microscopy and quantified. Flow cytometry showed that 1C5 and 2C2 were Le(a) and Le(b) positive. 1C5 expressed Le(b) on O-linked, but not N-linked, glycans and only weakly on GLs. In contrast, 2C2 expressed Le(b) on N-, O-glycans, and GLs. Furthermore, both clones expressed Le(a) on N- and O-glycans but not on GLs. 2C2, but not 1C5, stained positively for Le(y) on N-linked glycans and GLs. Both clones, as well as the parental CHO-K1 cells, expressed Le(x) on GLs. A Le(b)-binding H....Continue Reading

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Citations

May 21, 2011·The Journal of Biological Chemistry·Nozomi IshijimaHitomi Mimuro
Aug 16, 2013·BMC Microbiology·Nelly Romani VestmanIngegerd Johansson
Nov 19, 2013·Immunology Letters·Karien BloemYvette van Kooyk
Mar 10, 2011·Journal of Physics. Condensed Matter : an Institute of Physics Journal·S MichalikL K Varga
Jan 9, 2010·Analytical Chemistry·Joseph Sherma

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