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

• Published in: Microbes and infection Vol. 8; no. 7; pp. 1813 - 1825
• Main Authors: 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 A.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier SAS 01.06.2006; Amsterdam Elsevier; Paris
• Subjects: 3-D cell culture; Bacterial Adhesion; Bacterial diseases; Bacterial diseases of the digestive system and abdomen; Bacteriology; Biological and medical sciences; Cell Culture Techniques; Colon - cytology; Colon - microbiology; Cytoplasm - microbiology; Fundamental and applied biological sciences. Psychology; HT-29; HT29 Cells; Human bacterial diseases; Humans; Immunohistochemistry; Infectious diseases; Interleukin-8 - biosynthesis; Intestinal Mucosa - cytology; Intestinal Mucosa - microbiology; Medical sciences; Microbiology; Microscopy, Electron, Scanning; Miscellaneous; Organoids - chemistry; Organoids - cytology; Organoids - microbiology; Organoids - ultrastructure; Organotypic model; Salmonella typhimurium; Salmonella typhimurium - growth & development; Salmonella typhimurium - pathogenicity; Salmonella typhimurium - physiology; SPI-I; Organotypic model; HT-29; Salmonella typhimurium; 3-D cell culture; SPI-I; Human; Cell culture; Digestive system; Gut; Early stage; HT-29: Salmonella typhimurium; Epithelium; Infection; Bacteriosis; Bacteria; Models; Salmonellosis; Colon; Enterobacteriaceae
• ISSN: 1286-4579; 1769-714X
• DOI: 10.1016/j.micinf.2006.02.020

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.