C L Wells1, R P Jechorek, S L Erlandsen. 1. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455-0374.
Abstract
OBJECTIVE: To clarify the effect of bile salts on internalization of enteric bacteria by intestinal epithelial cells. DESIGN: Randomized study. SETTING: Research laboratory. SUBJECTS: Cultured human intestinal epithelial cells, namely HT-29 cells. INTERVENTIONS: The effect of bile was studied by adding bile during the time period in which bacterial cells were permitted to interact with enterocytes. In subsequent experiments, bile was added to the culture medium used to grow bacteria, and bacterial cells were washed before adding bacteria to enterocytes. Three different concentrations of three different bile preparations were tested. MEASUREMENTS AND MAIN RESULTS: Salmonella typhimurium and Proteus mirabilis were each incubated with HT-29 cells for 1 hr; the numbers of internalized bacteria were subsequently quantified following enterocyte lysis. The presence of bile during bacteria-enterocyte incubation had no effect on the numbers of internalized bacteria. However, if S. typhimurium or P. mirabilis were grown in the presence of bile, these washed bacterial cells were generally internalized by HT-29 cells in significantly fewer numbers, compared with bacterial cells grown in medium without bile supplementation. Enterocyte viability and morphologic ultrastructure did not appear to be affected by the presence of bile itself, or by the interaction with bacterial cells that had been cultivated in unsupplemented medium or in bile-supplemented medium. CONCLUSIONS: Exposure to bile during bacterial growth resulted in bacterial cells with decreased invasiveness for cultured intestinal epithelial cells. This observation is consistent with previous in vivo studies of obstructive jaundice, where the absence of bile in the intestinal lumen, not bile duct ligation, appeared to facilitate bacterial translocation in obstructed animals. Thus, the presence of bile in the intestinal lumen may decrease bacterial translocation by a mechanism that involves decreased epithelial internalization of enteric bacteria.
OBJECTIVE: To clarify the effect of bile salts on internalization of enteric bacteria by intestinal epithelial cells. DESIGN: Randomized study. SETTING: Research laboratory. SUBJECTS: Cultured human intestinal epithelial cells, namely HT-29 cells. INTERVENTIONS: The effect of bile was studied by adding bile during the time period in which bacterial cells were permitted to interact with enterocytes. In subsequent experiments, bile was added to the culture medium used to grow bacteria, and bacterial cells were washed before adding bacteria to enterocytes. Three different concentrations of three different bile preparations were tested. MEASUREMENTS AND MAIN RESULTS:Salmonella typhimurium and Proteus mirabilis were each incubated with HT-29 cells for 1 hr; the numbers of internalized bacteria were subsequently quantified following enterocyte lysis. The presence of bile during bacteria-enterocyte incubation had no effect on the numbers of internalized bacteria. However, if S. typhimurium or P. mirabilis were grown in the presence of bile, these washed bacterial cells were generally internalized by HT-29 cells in significantly fewer numbers, compared with bacterial cells grown in medium without bile supplementation. Enterocyte viability and morphologic ultrastructure did not appear to be affected by the presence of bile itself, or by the interaction with bacterial cells that had been cultivated in unsupplemented medium or in bile-supplemented medium. CONCLUSIONS: Exposure to bile during bacterial growth resulted in bacterial cells with decreased invasiveness for cultured intestinal epithelial cells. This observation is consistent with previous in vivo studies of obstructive jaundice, where the absence of bile in the intestinal lumen, not bile duct ligation, appeared to facilitate bacterial translocation in obstructed animals. Thus, the presence of bile in the intestinal lumen may decrease bacterial translocation by a mechanism that involves decreased epithelial internalization of enteric bacteria.
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