Estudo de genes de Candida albicans com função desconhecida quanto à formação de biofilme, características biológicas e interação patógeno- hospedeiro

• Autor(a) principal: Costa, Anna Carolina Borges Pereira da [UNESP]
• Data de Publicação: 2015
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/136649; http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/28-03-2016/000860821.pdf
• topic: Candida albicans; Biofilme; Antimicóticos; Patogenese

Candida albicans is an opportunistic fungi capable of causing superficial and systemic infections. Most of infections are mediated by biofilm formation which confers resistance to antifungal agents and immune system, but the mechanisms of biofilm development and pathogenicity were not thoroughly elucidated yet. In the presente study, 9 unknown function genes of C. albicans were selected among 34 mutant strains that presented altered phenotype for biofilm formation. The biofilms were formed on 96-well microtitle plates or on polystyrene disks and evaluated in different time intervals. Next, 4 complemented strains were constructed and evaluated for susceptibility to stressor agents, growth under nutrient limitation and filamentation tests. The biofilm architecture was analyzed by scanning electron microscopy (SEM). The biofilms were also assessed as the quantity of β-1,3-glucan and chitin. For the infection models, buccal epithelial cells (TR-146) were used for adherence, invasion and damage assays. The pathogenicity of the strains was evaluated in embrionated chicken eggs for 7 days, after inoculation of the strains. Planktonic cells and biofilms were submitted to antifungal tests with fluconazole, amphotericin B, and caspofungin. Quantitative polymerase chain reaction was performed to verify the expression of MRV8 and NDT80 genes in fungal cells in interaction with epithelial cells and MRV8, MRV1, and MRV6 genes in cells grown in biofilm. The results were submitted to the Student t test, ANOVA, Tukey's test, and Log-rank test (Mantel-Cox) (p < 0.05). Four complemented strains were constructed for the selected genes ORF19.823, ORF19.7170, ORF19.6847, and MRV8. The function of the ORF19.823 is still unknown, because the mutant strain did not show any significative phenotype for the tests performed. The mutant strain for the ORF19.7170 caused less damage on epithelial cells, but the result was not significant and the gene was dispensable...