Role of strain type, AGS cells and fetal calf serum in Helicobacter pylori adhesion and invasion assays.
In a human gastric biopsy specimen, 30% of adhering Helicobacter pylori strain AF4 (cagA and VacA positive) was associated with adhesion pedestals. In an AGS cell assay, only a few percent of this type I strain was found to be associated with adhesion pedestals. Nevertheless, a larger proportion of the type I strain was found to invade AGS cells (P < 0.03) and to attach with depressions in the AGS cell membrane (P < 0.03) than a type II strain (cagA and VacA negative). Incubation of AGS cells and H. pylori without adding fetal calf serum (FCS) to the culture medium increased actin accumulations (FITC-phalloidin stained) beneath adhering H. pylori, and decreased H. pylori invasion of AGS cells significantly (P < 0.01). However, no increase in the number of adhesion pedestals was observed by electron microscopy. Proteinase K treatment of FCS eliminated the H. pylori invasion promoting effect (P < 0.01). Our results suggest differences in the ability of H. pylori to induce adhesion pedestals in human gastric epithelial cells and in AGS cells, but a correlation between adhesion pedestal formation in vivo and H. pylori invasion in vitro can be speculated. In addition, H. pylori invasion into AGS cells was found to be mediated by proteins in FCS.
Petersen
A M
AM
Department of Gastrointestinal Infections and Laboratory for Electron Microscopy, Statens Serum Institsut, Copenhagen, Denmark.
Blom
J
J
Andersen
L P
LP
Krogfelt
K A
KA
D000818:Animals; D001422:Bacterial Adhesion; D001706:Biopsy; D001769:Blood; D002417:Cattle; D016895:Culture Media, Serum-Free; D004847:Epithelial Cells / Q000382:microbiology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique / Q000379:methods; D005753:Gastric Mucosa / Q000166:cytology / Q000382:microbiology* / Q000648:ultrastructure; D005839:Gentamicins; D016480:Helicobacter pylori / Q000145:classification / Q000472:pathogenicity / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D014407:Tumor Cells, Cultured; D014774:Virulence
D016895:Culture Media, Serum-Free; D005839:Gentamicins