CELL ASSOCIATION AND INVASION OF CACO-2 CELLS BY CAMPYLOBACTER-JEJUNI

Adherence and invasion studies were conducted in monolayers of Caco-2 cells. Three-day-old monolayers were inoculated with Campylobacter jejuni 81-176 at a bacterium/cell ratio of 1,000:1. Saturation studies demonstrated time- and dose-dependent saturation curves for C. jejuni cell association and invasion into Caco-2 cells. Electron microscopy revealed intracellular C.jejuni located within membrane-bound vacuoles. Cell association and invasion were inhibited by 0.3 and 0.5 M concentrations of various sugars, including D-glucose, D-mannose, and D-fucose. However, there was no inhibition with the corresponding L-sugars, indicating physiological specificity. The inhibition of cell association with phloridzin was less pronounced. There was no inhibition of bacterial entry with monodansylcadaverine or g-strophanthin, indicating that it was unlikely that coated-pit formation is important in the invasion of C.jejuni into Caco-2 cells. Furthermore, there was no inhibition with cytochalasin D, vincristine, or vinblastine. Inhibition of cell association was demonstrated at 4 degrees C. Significantly decreased cell association and invasion were seen in potassium-depleted cells. Treatment of eels with bromelain also caused reduction in the number of C.jejuni binding to cells. A nonmotile aflagellate variant of C.jejuni also showed reduced invasion. The results of this study are consistent with energy-dependent invasion mechanisms. The results do not support an endocytic method of invasion for C.jejuni into Caco-2 cells.