Some structures and processes of human epithelial cells involved in uptake of enterohemorrhagic Escherichia coli O157:H7 strains.
Several enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 isolated from patients with hemorrhagic colitis, ischemic colitis, or hemolytic uremic syndrome were all found to be able to invade certain human epithelial cell lines in vitro. Their ability to gain entry into epithelial cells was compared with those of known invasive Shigella flexneri and Salmonella typhi strains and the noninvasive E. coli strain HB101 in invasion assays utilizing gentamicin to kill extracellular bacteria. All EHEC strains under investigation were efficiently internalized into T24 bladder and HCT-8 ileocecal cells. In striking contrast to shigellae, the same EHEC strains were not taken up into human embryonic intestinal INT407 cells or HEp-2 cells any more than the noninvasive E. coli strain HB101. The mechanism(s) of EHEC internalization was characterized by comparing the invasion efficiencies in the absence and presence of a variety of inhibitors acting on structures and processes of prokaryotic or eukaryotic cells. Also, wild-type, plasmid-containing EHEC strains were compared with their plasmid-cured isogenic derivative strains to determine if plasmid genes affect invasion ability. Plasmid-cured EHEC invaded as well as wild-type EHEC, indicating that invasion ability is chromosomally encoded. Inhibition of bacterial protein synthesis by simultaneous addition of bacteria and chloramphenicol to the monolayer blocked EHEC uptake dramatically, suggesting the presence of an invasion protein(s) with a short half-life. Studies utilizing inhibitors which act on eukaryotic cells demonstrated a strong dependence on microfilaments in the process of uptake of all EHEC strains into both T24 and HCT-8 cells. In general, depolymerization of microtubules as well as inhibition of receptor-mediated endocytosis reduced the efficiency of EHEC invasion of T24 cells, whereas interference with endosome acidification reduced EHEC entry into only HCT-8 cells. Taxol-induced stabilization of microtubules did not inhibit internalization into T24 cells or into the HCT-8 cell line. In marked contrast, the ability of S. typhi Ty2 to invade either cell line was inhibited only by depolymerization of microfilaments. In addition to the cell line specificity of EHEC invasion, not all EHEC strains displayed uniform behavior in the presence of inhibitors, suggesting the existence of variant uptake pathways in different strains. Most importantly, previous reports of the inability of EHEC to invade INT407 or HEp-2 cell lines support the currently held belief that EHEC strains are noninvasive.(ABSTRACT TRUNCATED AT 400 WORDS)
Oelschlaeger
T A
TA
Department of Bacterial Diseases, Walter Reed Army Institute of Research, Washington, D.C. 20307-5100.
Barrett
T J
TJ
Kopecko
D J
DJ
D008841:Actin Cytoskeleton / Q000502:physiology; D001426:Bacterial Proteins / Q000096:biosynthesis; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D002701:Chloramphenicol / Q000494:pharmacology; D002966:Clathrin / Q000502:physiology; D003092:Colitis / Q000382:microbiology*; D003593:Cytoplasm / Q000382:microbiology; D004269:DNA, Bacterial / Q000096:biosynthesis; D004705:Endocytosis; D004848:Epithelium / Q000382:microbiology*; D004926:Escherichia coli / Q000254:growth & development / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology* / Q000473:pathology; D006463:Hemolytic-Uremic Syndrome / Q000382:microbiology*; D006470:Hemorrhage; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008870:Microtubules / Q000502:physiology; D009675:Novobiocin / Q000494:pharmacology; D012329:RNA, Bacterial / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000502:physiology; D012294:Rifamycins / Q000494:pharmacology; D012485:Salmonella typhi / Q000254:growth & development; D012763:Shigella flexneri / Q000254:growth & development
D001426:Bacterial Proteins; D002966:Clathrin; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D011956:Receptors, Cell Surface; D012294:Rifamycins; D009675:Novobiocin; D002701:Chloramphenicol; C008542:monodansylcadaverine; D002103:Cadaverine