PT  - JOURNAL ARTICLE
AU  - Blessing O. Anonye
AU  - Jack Hassall
AU  - Jamie Patient
AU  - Usanee Detamornrat
AU  - Afnan M. Aladdad
AU  - Stephanie Schüller
AU  - Felicity R.A.J. Rose
AU  - Meera Unnikrishnan
TI  - Probing <em>Clostridium difficile</em> infection in innovative human gut cellular models
AID  - 10.1101/269035
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 269035
4099  - http://biorxiv.org/content/early/2019/02/08/269035.short
4100  - http://biorxiv.org/content/early/2019/02/08/269035.full
AB  - Interactions of anaerobic gut bacteria, such as Clostridium difficile, with the intestinal mucosa have been poorly studied due to challenges in culturing anaerobes with the oxygen-requiring gut epithelium. Although gut colonization by C. difficile is a key determinant of disease outcome, precise mechanisms of mucosal attachment and spread remain unclear. Here, using human gut epithelial monolayers co-cultured within dual environment chambers, we demonstrate that C. difficile adhesion to gut epithelial cells is accompanied by a gradual increase in bacterial numbers. Prolonged infection causes redistribution of actin and loss of epithelial integrity, accompanied by production of C. difficile spores, toxins and bacterial filaments. This 2-D dual chamber system was used to examine C. difficile interactions with the commensal Bacteroides dorei, and interestingly, C. difficile growth is significantly reduced in presence of B. dorei. Furthermore, in novel multilayer and 3-D gut models containing a myofibroblast layer, C. difficile adheres more efficiently to epithelial cells, as compared to the 2-D model, leading to a quicker destruction of the epithelium. Our study describes new controlled environment human gut models that enable host-anaerobe and pathogen-commensal interaction studies in vitro.