Vaginal Epithelium Model

The reconstituted human vaginal epithelium (RHVE) resembles the multilayer human vaginal mucosa and is commercialized by Skinethic Laboratory (Nice, France). It is based on a cell line which was obtained by culturing transformed human keratinocytes of the cell line A431 derived from a vulval epidermoid carcinoma (Rosdy et al., 1986). The RHVE model was valid for evaluating the phenotype of mutants in the agglutinin-like sequence (ALS) family in C. albicans. The expression of ALSs proteins in the in vitro RHVE was compared with in vivo expression in a murine vaginitis model (Cheng et al., 2005). The RHVE model has been also used to study the epithelial cytokine response induced by C. albicans and the role of the Secreted Aspartyl Proteinases (Saps), important virulence factors of this fungus. Saps cause tissue damage and the different damaging potential of each Sap correlates with an epithelium-induced pro-inflammatory cytokine response, which could be crucial in the control of the vaginal infection by C. albicans (Schaller et al., 2005). Pro-inflammatory and other chemoattractive cytokines generated by RHVE cells were detected and quantified by quantitative reverse transcription-PCR and fluorescence-activated cell sorter (FACS) analyses.

The expression of pro-inflammatory cytokines, chemokines, and ß-defensins-2 was also measured using another immortalized vaginal epithelial cell line, PK E6/ E7 (Pivarcsi et al., 2005). The expression of inflammatory mediators and defensins induced in the presence of compounds from different pathogenic microorganisms, among them heat-killed C. albicans, was quantified by Q-RT-PCR, flow cytometry and ELISA (Pivarcsi et al., 2005).

Anti- Candida activity of vaginal epithelial cells was identified as a possible factor of local immunity in vulvovaginal candidiasis by using different cell models (Barousse et al., 2001; Barousse et al., 2005; Steele et al., 1999a, b). Co-inhabitants at the vaginal site of infection are bacteria of the genus Lactobacillus spp. which constitute the protective part of the microbiota while Gardnerella vaginalis is the most common bacteria associated with bacterial vaginosis. Little is known about the impact of these - and other - members of the microbial community on the virulence of C. albicans. A further development of the vaginal in vitro system could certainly be the addition of members of the microbiota to model the natural micro-environment.

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