Comparison of the Caco-2, HT-29 and the mucus-secreting HT29-MTX intestinal cell models to investigate Salmonella adhesion and invasion

J Microbiol Methods. 2013 Sep;94(3):274-9. doi: 10.1016/j.mimet.2013.06.027. Epub 2013 Jul 5.

Abstract

Human intestinal cell models are widely used to study host-enteric pathogen interactions, with different cell lines exhibiting specific characteristics and functions in the gut epithelium. In particular, the presence of mucus may play an important role in adhesion and invasion of pathogens. The aim of this study was to evaluate the suitability of the mucus-secreting HT29-MTX intestinal epithelial cell model to test adhesion and invasion of Salmonella strains and compare with data obtained with the more commonly used Caco-2 and HT-29 models. Adhesion of Salmonella to HT29-MTX cell model was significantly higher, likely due to high adhesiveness to mucins present in the native human mucus layer covering the whole cell surface, compared to the non- and low-mucus producing Caco-2 and HT-29 cell models, respectively. In addition, invasion percentages of some clinical Salmonella strains to HT29-MTX cultures were remarkably higher than to Caco-2 and HT-29 cells suggesting that these Salmonellae have subverted the mucus to enhance pathogenicity. The transepithelial electrical resistances of the infected HT29-MTX cell model decreased broadly and were highly correlated with invasion ability of the strain. Staining of S. Typhimurium-infected cell epithelium confirmed the higher invasion by Salmonella and subsequent disruption of tight junctions of HT29-MTX cell model compared with the Caco-2 and HT-29 cell models. Data from this study suggest that the HT29-MTX cell model, with more physiologically relevant characteristics with the mucus layer formation, could be better suited for studying cells-pathogens interactions.

Keywords: 4′,6′-diamidino-2-phenylindole; DAPI; DMEM; Dulbecco's Modified Eagle medium; FBS; HT29-MTX cells; Invasion; Mucus; RPMI; Roswell Park Memorial Institute 1640 medium; Salmonella; TER; TJs; TLRs; TRITC-phalloidin; Tight junctions; Toll-like receptors; fetal bovine serum; tetramethyl-rhodamine B isothiocyanate-phalloidin; tight junctions; transepithelial electrical resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Bacterial Adhesion / physiology*
  • Caco-2 Cells
  • Electric Impedance
  • HT29 Cells
  • Humans
  • Intestinal Mucosa / cytology*
  • Models, Biological*
  • Salmonella / physiology*
  • Tight Junctions