Entamoeba histolytica interactions with polarized human intestinal Caco-2 epithelial cells.
To model the initial pathogenic effects of Entamoeba histolytica trophozoites on intestinal epithelial cells, the interactions of E. histolytica HM1-IMSS trophozoites with polarized human intestinal Caco-2 cell monolayers grown on permeabilized filters were examined. Trophozoites, when incubated with the apical surface of the monolayers at 37 degrees C, induced a rapid decrease in transepithelial resistance over 15 to 60 min. The transmonolayer resistance response was not associated with changes in short-circuit current but was associated with an increase in mannitol flux, suggesting that the drop in resistance reflected a nonselective increase in epithelial permeability rather than stimulation of electrogenic ion transport. This response preceded the earliest detection of morphologic disruption of monolayer integrity by light or electron microscopy. Apical injury to the monolayer was detected by ultrastructural studies which revealed a loss of brush border in regions of contact between epithelial cells and amebas and by chromium release assays where a small increase in the apical release of 51Cr from the monolayer (6% over background) was observed. The transmonolayer resistance response was inhibited when the temperature was reduced to 4 degrees C and by addition of cytochalasin D (1 microgram/ml) to the medium at concentrations that did not directly affect transmonolayer resistance. Application of amebic lysates or medium conditioned by coincubation of amebas with Caco-2 monolayers failed to lower transmonolayer resistance, suggesting that this effect was not mediated by soluble amebic cytotoxins. Polarized Caco-2 monolayers grown on permeable filters provide a useful model for studying the initial interactions of E. histolytica trophozoites with intestinal epithelial cells.
Li
E
E
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Stenson
W F
WF
Kunz-Jenkins
C
C
Swanson
P E
PE
Duncan
R
R
Stanley
S L
SL
D000818:Animals; D001692:Biological Transport; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D002463:Cell Membrane Permeability; D016764:Cell Polarity; D015638:Cytochalasin D / Q000494:pharmacology; D004594:Electrophysiology; D004748:Entamoeba histolytica / Q000254:growth & development*; D004847:Epithelial Cells; D004848:Epithelium / Q000469:parasitology; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000469:parasitology*; D007785:Lactose / Q000494:pharmacology; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008353:Mannitol / Q000378:metabolism; D010539:Permeability
D015638:Cytochalasin D; D008353:Mannitol; D007930:Leucine; D007785:Lactose; C024974:E 64