Three-dimensional organotypic models of human colonic epithelium to study the early stages of enteric salmonellosis

Kerstin Höner zu Bentrup, Rajee Ramamurthy, C. Mark Ott, Kamal Emami, Mayra Nelman-Gonzalez, James W. Wilson, Emily G. Richter, Thomas J. Goodwin, J. Stephen Alexander, Duane L. Pierson, Neal Pellis, Kent L. Buchanan, Cheryl Nickerson

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

In vitro cell culture models used to study how Salmonella initiates disease at the intestinal epithelium would benefit from the recognition that organs and tissues function in a three-dimensional (3-D) environment and that this spatial context is necessary for development of cultures that more realistically resemble in vivo tissues/organs. Our aim was to establish and characterize biologically meaningful 3-D models of human colonic epithelium and apply them to study the early stages of enteric salmonellosis. The human colonic cell line HT-29 was cultured in 3-D and characterized by immunohistochemistry, histology, and scanning electron microscopy. Wild-type Salmonella typhimurium and an isogenic SPI-1 type three secretion system (TTSS) mutant derivative (invA) were used to compare the interactions with 3-D cells and monolayers in adherence/invasion, tissue pathology, and cytokine expression studies. The results showed that 3-D culture enhanced many characteristics normally associated with fully differentiated, functional intestinal epithelia in vivo, including better organization of junctional, extracellular matrix, and brush-border proteins, and highly localized mucin production. Wild-type Salmonella demonstrated increased adherence, but significantly lower invasion for 3-D cells. Interestingly, the SPI-I TTSS mutant showed wild-type ability to invade into the 3-D cells but did not cause significant structural changes to these cells. Moreover, 3-D cells produced less interleukin-8 before and after Salmonella infection. These results suggest that 3-D cultures of human colonic epithelium provide valuable alternative models to study human enteric salmonellosis with potential for novel insight into Salmonella pathogenesis.

Original languageEnglish (US)
Pages (from-to)1813-1825
Number of pages13
JournalMicrobes and Infection
Volume8
Issue number7
DOIs
StatePublished - Jun 2006
Externally publishedYes

Fingerprint

Somatostatin-Secreting Cells
Salmonella Infections
Epithelium
Salmonella
Intestinal Mucosa
Mucins
Salmonella typhimurium
Microvilli
Interleukin-8
Electron Scanning Microscopy
Extracellular Matrix
Histology
Cell Culture Techniques
Immunohistochemistry
Pathology
Cytokines
Cell Line
Proteins
Type I Secretion Systems

Keywords

  • 3-D cell culture
  • HT-29
  • Organotypic model
  • Salmonella typhimurium
  • SPI-I

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Three-dimensional organotypic models of human colonic epithelium to study the early stages of enteric salmonellosis. / Höner zu Bentrup, Kerstin; Ramamurthy, Rajee; Ott, C. Mark; Emami, Kamal; Nelman-Gonzalez, Mayra; Wilson, James W.; Richter, Emily G.; Goodwin, Thomas J.; Alexander, J. Stephen; Pierson, Duane L.; Pellis, Neal; Buchanan, Kent L.; Nickerson, Cheryl.

In: Microbes and Infection, Vol. 8, No. 7, 06.2006, p. 1813-1825.

Research output: Contribution to journalArticle

Höner zu Bentrup, K, Ramamurthy, R, Ott, CM, Emami, K, Nelman-Gonzalez, M, Wilson, JW, Richter, EG, Goodwin, TJ, Alexander, JS, Pierson, DL, Pellis, N, Buchanan, KL & Nickerson, C 2006, 'Three-dimensional organotypic models of human colonic epithelium to study the early stages of enteric salmonellosis', Microbes and Infection, vol. 8, no. 7, pp. 1813-1825. https://doi.org/10.1016/j.micinf.2006.02.020
Höner zu Bentrup, Kerstin ; Ramamurthy, Rajee ; Ott, C. Mark ; Emami, Kamal ; Nelman-Gonzalez, Mayra ; Wilson, James W. ; Richter, Emily G. ; Goodwin, Thomas J. ; Alexander, J. Stephen ; Pierson, Duane L. ; Pellis, Neal ; Buchanan, Kent L. ; Nickerson, Cheryl. / Three-dimensional organotypic models of human colonic epithelium to study the early stages of enteric salmonellosis. In: Microbes and Infection. 2006 ; Vol. 8, No. 7. pp. 1813-1825.
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AU - Höner zu Bentrup, Kerstin

AU - Ramamurthy, Rajee

AU - Ott, C. Mark

AU - Emami, Kamal

AU - Nelman-Gonzalez, Mayra

AU - Wilson, James W.

AU - Richter, Emily G.

AU - Goodwin, Thomas J.

AU - Alexander, J. Stephen

AU - Pierson, Duane L.

AU - Pellis, Neal

AU - Buchanan, Kent L.

AU - Nickerson, Cheryl

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N2 - In vitro cell culture models used to study how Salmonella initiates disease at the intestinal epithelium would benefit from the recognition that organs and tissues function in a three-dimensional (3-D) environment and that this spatial context is necessary for development of cultures that more realistically resemble in vivo tissues/organs. Our aim was to establish and characterize biologically meaningful 3-D models of human colonic epithelium and apply them to study the early stages of enteric salmonellosis. The human colonic cell line HT-29 was cultured in 3-D and characterized by immunohistochemistry, histology, and scanning electron microscopy. Wild-type Salmonella typhimurium and an isogenic SPI-1 type three secretion system (TTSS) mutant derivative (invA) were used to compare the interactions with 3-D cells and monolayers in adherence/invasion, tissue pathology, and cytokine expression studies. The results showed that 3-D culture enhanced many characteristics normally associated with fully differentiated, functional intestinal epithelia in vivo, including better organization of junctional, extracellular matrix, and brush-border proteins, and highly localized mucin production. Wild-type Salmonella demonstrated increased adherence, but significantly lower invasion for 3-D cells. Interestingly, the SPI-I TTSS mutant showed wild-type ability to invade into the 3-D cells but did not cause significant structural changes to these cells. Moreover, 3-D cells produced less interleukin-8 before and after Salmonella infection. These results suggest that 3-D cultures of human colonic epithelium provide valuable alternative models to study human enteric salmonellosis with potential for novel insight into Salmonella pathogenesis.

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