Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia for the study of host-pathogen...

This protocol describes the setup, maintenance, and characteristics of models of oral and vaginal candidiasis based on well-established three-dimensional organotypic tissues of human oral and vaginal mucosa. Infection experiments are highly reproducible and can be used for the direct analysis of pathogen/epithelial cell interactions. Using the models, the several stages of infection by wild-type Candida albicans strains, the consequence of gene disruption of putative virulence factors in mutant cells, and the evaluation of the host immune response can be evaluated by histologic, biochemical, and molecular methods. As such, the models provide clear answers regarding protein and gene expression that are not complicated by nonepithelial factors. To study the impact of several host components, the mucosal infection models can be supplemented with immune cells, saliva, and probiotic bacteria, which might be relevant for host defense. It requires at least 3 days to be established and can be maintained thereafter for 2 to 4 days.