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

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.

journal_name

Microbes Infect

journal_title

Microbes and infection

authors

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 CA

doi

10.1016/j.micinf.2006.02.020

subject

Has Abstract

pub_date

2006-06-01 00:00:00

pages

1813-25

issue

7

eissn

1286-4579

issn

1769-714X

pii

S1286-4579(06)00143-2

journal_volume

8

pub_type

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