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Influence of fluconazole in immune response to infection by Candida albicans on in vitro model of oral epithelium


In the last years, factors such as increased incidence of invasive fungal diseases, as well as the emergence of drug resistant isolates, have increased the interest of such drugs influence not only on the pathogens, but also on the host immune response. Considering the limited knowledge about the influence of salivary concentrations of antifungal therapy used for the oral candidiasis treatment on the interaction between pathogen and host, the aim of this study is to evaluate the in vitro immune response of human oral epithelium infected with Candida albicans biofilm and, separately, the interaction between C. albicans and macrophages, both in the presence of plasma and saliva fluconazole concentration, respectively. Two clinical isolates (P01, P34) and a reference strain (ATCC 90028) of C. albicans will be used. The epithelium will be inoculated with a standardized C. albicans suspension (2x106 cels./mL) and the RPMI 1640 with 2.56 mg/mL of fluconazole, will be added. After 24 hours, the assessment of interleukins and secretory aspartyl proteases (SAPs) gene expression will be performed by quantitative PCR (qPCR) using specific primer pairs. Quantification of epithelial damage will be assessed by detecting lactate dehydrogenase (LDH) release. The interaction between C. albicans and macrophages will be performed inside cell culture plates. Phagocytosis and intracellular killing of C. albicans will be made by plating and CFU counting, and also it will be quantified the nitric oxide and interleukins release by macrophages. Complementary analyzes of tissue and the biofilm structure will be performed by laser confocal microscopy, in order to verify the fungal invasion and biovolume, structure and roughness of the biofilm. At the end of the study, data will be analyzed according to their distribution and appropriate statistical tests will be applied. (AU)

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(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BERTOLINI, M. M.; XU, H.; SOBUE, T.; NOBILE, C. J.; DEL BEL CURY, A. A.; DONGARI-BAGTZOGLOU, A. Candida-streptococcal mucosal biofilms display distinct structural and virulence characteristics depending on growth conditions and hyphal morphotypes. Molecular Oral Microbiology, v. 30, n. 4, p. 307-322, AUG 2015. Web of Science Citations: 18.

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