T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis

James P. Nataro, Susan Hicks, Alan D. Phillips, Pablo A. Vial, Cynthia Louise Sears

Research output: Contribution to journalArticle

Abstract

Enteroaggregative Escherichia coli (EAEC) is an important cause of persistent diarrhea in many developing parts of the world, yet the pathogenetic mechanisms of EAEC diarrhea are unknown. Experiments with animal models suggest that EAEC strains damage the intestinal mucosa, and a putative cytotoxin has been described. To characterize the mucosal effects of EAEC, we studied strain 042, which we have shown to cause diarrhea in adult volunteers. Strain 042 was incubated in an in vitro organ culture model with biopsy-derived normal intestinal mucosa from pediatric patients. Strain 042 adhered strongly to samples of jejunal, ileal, and colonic mucosa. In addition, scanning electron microscopic examination of in vitro-infected intestinal biopsies revealed cytotoxic effects marked by exfoliation of mucosal epithelial cells. To develop an in vitro model to study these effects, we incubated 042 with polarized monolayers of the human intestinal epithelial cell lines Caco-2 and T84. Strain 042 adhered strongly to T84 cells but not to Caco-2 cells. T84 cells infected with 042 displayed marked toxic effects, most prominently in areas where bacteria were adhering. The apical membrane of damaged cells exhibited vesiculation and shedding of microvilli. The cytoplasm of affected cells displayed subnuclear vacuolization, and in some cases, nuclei of affected cells became separated from the surrounding cytoplasm. Severely affected cells ruptured, releasing their nuclei. Vacuolated remnant cells were seen throughout the monolayer. Strain 042 was not internalized by T84 cells. We concluded that EAEC strain 042 alters intestinal cell morphology, ultimately leading to cell death. Although the factor(s) required for this effect remains to be elucidated, T84 cells may serve as a valuable model in EAEC pathogenesis studies.

Original languageEnglish (US)
Pages (from-to)4761-4768
Number of pages8
JournalInfection and Immunity
Volume64
Issue number11
StatePublished - 1996

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Cell Culture Techniques
Escherichia coli
Diarrhea
Intestinal Mucosa
Cytoplasm
Epithelial Cells
Biopsy
Caco-2 Cells
Poisons
Organ Culture Techniques
Cytotoxins
Microvilli
Cell Nucleus
Volunteers
Mucous Membrane
Cell Death
Animal Models
Cell Membrane
Electrons
Pediatrics

ASJC Scopus subject areas

  • Immunology

Cite this

Nataro, J. P., Hicks, S., Phillips, A. D., Vial, P. A., & Sears, C. L. (1996). T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis. Infection and Immunity, 64(11), 4761-4768.

T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis. / Nataro, James P.; Hicks, Susan; Phillips, Alan D.; Vial, Pablo A.; Sears, Cynthia Louise.

In: Infection and Immunity, Vol. 64, No. 11, 1996, p. 4761-4768.

Research output: Contribution to journalArticle

Nataro, JP, Hicks, S, Phillips, AD, Vial, PA & Sears, CL 1996, 'T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis', Infection and Immunity, vol. 64, no. 11, pp. 4761-4768.
Nataro, James P. ; Hicks, Susan ; Phillips, Alan D. ; Vial, Pablo A. ; Sears, Cynthia Louise. / T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis. In: Infection and Immunity. 1996 ; Vol. 64, No. 11. pp. 4761-4768.
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