i(3)so3-galactosylceramide and Stomach-Neoplasms

Helicobacter pylori adhere to Kato III and Hela S3 cells in monolayer cultures. To explore whether cell surface glycoconjugates on these two cell lines mediate binding of H. pylori, various carbohydrates, glycoproteins, and glycolipids were tested to inhibit H.pylori cell adhesion. The adhesion was measured (i) with a urease-based assay and (ii) by cells stained with fluorescein. Sodium periodate and sialidase treatment (but not alpha- or beta-galactosidase, heparitinase,lysozyme, or trypsin) inhibited H. pylori binding to both cell lines. Sulfatides and sulfated glycoconjugates (50 microg/ml) but not heparin or a number of simple carbohydrates inhibited binding (1 mg/ml). The two H.pylori strains studied (CCUG 17874 and strain 25) showed high binding of soluble 125I-labeled heparin and other sulfated carbohydrate compounds.

Topics: Bacterial Adhesion; Carbohydrate Metabolism; Glycoconjugates; HeLa Cells; Helicobacter pylori; Heparin; Humans; Neuraminidase; Periodic Acid; Stomach Neoplasms; Sulfoglycosphingolipids; Tumor Cells, Cultured

Immunohistochemical staining using a specific monoclonal antibody against sulfatide was performed to examine cellular localization of sulfatides in normal human gastric mucosa and in various types of gastric carcinoma. In normal gastric mucosa without Helicobacter pylori infection, both epithelial and glandular cells were densely stained with anti-sulfatide antibody. Sulfatide staining was more abundant in the apical area of normal epithelium compared with cytosol or basolateral areas. This tendency was stronger in the antrum than the gastric body and was more intensified in glandular cells of the pyloric glands. The levels of sulfatide expression were decreased in papillary and well-differentiated adenocarcinomas compared with normal mucosa, were markedly attenuated in moderately differentiated adenocarcinomas, and were very low in poorly differentiated adenocarcinomas. The polarity of the stains was preserved in gastric cancers that exhibited differentiated tissue organization. The levels of sulfatide expression were highly variable in signet ring carcinoma cells. The cancer cells that expressed sulfatides did not show any polarity of staining.

Topics: Adenocarcinoma; Adenocarcinoma, Papillary; Adolescent; Adult; Aged; Aged, 80 and over; Carcinoma, Signet Ring Cell; Female; Gastric Mucosa; Humans; Immunohistochemistry; Male; Middle Aged; Stomach Neoplasms; Sulfoglycosphingolipids

We have demonstrated that clinical isolates of Helicobacter pylori preferentially bind to sulfatides (I3SO3-GalCer) and GM3 gangliosides (II3NeuAcLacCer), two predominant acidic glycosphingolipids in the human gastric mucosa, on thin-layer chromatography plates. However, it has not yet been clarified that these glycospingolipids truly serve as adhesion receptors for H. pylori in live cells. In this study, we used a gastric cancer cell line, KATO III, as a cellular model of H. pylori adhesion and examined the role of sulfatides in attachment. The adhesion of H. pylori (i.e., a standard strain of H. pylori, NCTC 11637) to KATO III cells and the effects of various substances on this adhesion were monitored and semiquantitated by flow cytometric analysis. Sulfated glycoconjugates, such as heparin and gastric mucin, significantly inhibited H. pylori adhesion to KATO III cells. Membrane preparations from KATO III cells strongly inhibited this adhesion. In the membrane preparations, sulfatides were present as a major acidic glycosphinoglipid. With the exception of sulfatides, no distinct adhesion of H. pylori to glycospingolipids from KATO III cells were observed. Moreover, H. pylori did not bind to any membrane proteins of KATO III cells. Finally, a monoclonal anti-sulfatide antibody markedly reduced H. pylori adhesion to KATO III cells. These results suggest that sulfatides, and possibly related sulfated compounds, serve as a major receptor for cell adhesion by H. pylori.

Topics: Bacterial Adhesion; Glycosphingolipids; Helicobacter pylori; Humans; Stomach Neoplasms; Sulfoglycosphingolipids; Tumor Cells, Cultured