Surface hydrophobicity changes of two Candida albicans serotype B mnn4delta mutants.
ABSTRACT Cell surface hydrophobicity (CSH) of Candida species enhances virulence by promoting adhesion to host tissues. Biochemical analysis of yeast cell walls has demonstrated that the most significant differences between hydrophobic and hydrophilic yeasts are found in the acid-labile fraction of Candida albicans phosphomannoprotein, suggesting that this fraction is important in the regulation of the CSH phenotype. The acid-labile fraction of C. albicans is unique among fungi, in that it is composed of an extended polymer of beta-1,2-mannose linked to the acid-stable region of the N-glycan by a phosphodiester bond. C. albicans serotype A and B strains both contain a beta-1,2-mannose acid-labile moiety, but only serotype A strains contain additional beta-1,2-mannose in the acid-stable region. A knockout of the C. albicans homolog of the Saccharomyces cerevisiae MNN4 gene was generated in two serotype B C. albicans patient isolates by using homologous gene replacement techniques, with the anticipation that they would be deficient in the acid-labile fraction and, therefore, demonstrate perturbed CSH. The resulting mnn4delta-deficient derivative has no detectable phosphate-linked beta-1,2-mannose in its cell wall, and hydrophobicity is increased significantly under conditions that promote the hydrophilic phenotype. The mnn4delta mutant also demonstrates an unanticipated perturbation in the acid-stable mannan fraction. The present study reports the first genetic knockout constructed in a serotype B C. albicans strain and represents an important step for dissecting the regulation of CSH.
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ABSTRACT: The Candida antigen CR3-RP (complement receptor 3-related protein) is supposed to be a 'mimicry' protein because of its ability to bind antibody directed against the alpha subunit of the mammalian CR3 (CD11b/CD18). This study aimed to (i) investigate the specific humoral isotypic response to immunization with CR3-RP in vivo in a rabbit animal model, and (ii) determine the role of CR3-RP in the adherence of Candida albicans in vitro using the model systems of buccal epithelial cells (BECs) and biofilm formation. The synthetic C. albicans peptide DINGGGATLPQ corresponding to 11 amino-acids of the CR3-RP sequence DINGGGATLPQALXQITGVIT, determined by N-terminal sequencing, was used for immunization of rabbits to obtain polyclonal anti-CR3-PR serum and for subsequent characterization of the humoral isotypic response of rabbits. A significant increase of IgG, IgA and IgM anti-CR3-RP specific antibodies was observed after the third (P<0.01) and the fourth (P<0.001) immunization doses. The elevation of IgA levels suggested peptide immunomodulation of the IgA1 subclass, presumably in coincidence with Candida epithelial adherence. Blocking CR3-RP with polyclonal anti-CR3-RP serum reduced the ability of Candida to adhere to BECs, in comparison with the control, by up to 35 % (P<0.001), and reduced biofilm formation by 28 % (P<0.001), including changes in biofilm thickness and integrity detected by confocal laser scanning microscopy. These properties of CR3-RP suggest that it has potential for future vaccine development.Journal of Medical Microbiology 12/2008; 57(Pt 12):1466-72. DOI:10.1099/jmm.0.2008/001479-0 · 2.27 Impact Factor
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ABSTRACT: Glycosyltransferases are specific enzymes that catalyse the transfer of monosaccharide moieties to biological substrates, including proteins, lipids and carbohydrates. These enzymes are present from prokaryotes to humans, and their glycoconjugate products are often vital for survival of the organism. Many glycosyltransferases found in fungal pathogens such as Cryptococcus neoformans do not exist in mammalian systems, making them attractive potential targets for selectively toxic agents. In this article, we present the features of this diverse class of enzymes, and review the fungal glycosyltransferases that are involved in synthesis of the cell wall, the cryptococcal capsule, glycoproteins and glycolipids. We specifically focus on enzymes that have been identified or studied in C. neoformans, and we consider future directions for research on glycosyltransferases in the context of this opportunistic pathogen.FEMS Yeast Research 07/2006; 6(4):499-512. DOI:10.1111/j.1567-1364.2006.00054.x · 2.44 Impact Factor
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ABSTRACT: The aim of the present paper is to evaluate the effect of the high molecular weight chitosan (HMWC) and of sodium alginate (NaAL) on surface hydrophobicity of Candida albicans and on adhesion of the yeast to epithelial cells and fibroblasts of different proceeding. For this study, a collection strain and seven isolates of C. albicans from saliva (patients with denture stomatitis) were grown in Sabouraud glucose agar supplemented with HMWC or NaAL or in absence of them (control). Hydrophobicity was determined by adhesion to hydrocarbons method using two organic media (xylene and chloroform). For adhesion experiments, aqueous suspensions of yeasts were contacted with solutions of biopolymers and different cells (rat and human fibroblasts and epithelial cells Hep-2). The quantification of adhesion was made by optical microscopy. Results: a decrease in hydrophobicity was observed in the presence of HMWC (44%) and of NaAL (82%) when chloroform was employed as organic medium, meanwhile the decreases were of 30% with HMWC and 19% with NaAL in the presence of xylene. Adhesion of C. albicans to epithelial cells and human fibroblasts decreased significantly with both biopolymers. In the case of rat fibroblasts, a decrease was observed only with NaAL. None of experiments showed significant differences associated to fibroblast type. Conclusions: biopolymers showed effectiveness in reducing hydrophobicity and adhesion of C. albicans to cells, which are important virulence factors related to colonization of the soft tissues of host or acrylic surfaces present in the oral system. Objetivo: Evaluar el efecto del quitosán de alto peso molecular (QAPM) y del alginato de sodio (NaAL) sobre la hidrofobicidad superficial de Candida albicans y la adhesión de esta levadura a células epiteliales y fibroblastos de distinto origen. Diseño del estudio: Para el estudio de la hidrofobicidad, las levaduras (n=7) se hicieron crecer en agar glucosado de Sabouraud suplementado con QAPM o NaAL o en ausencia de los mismos (controles). La determinación de la hidrofobicidad se realizó por el método de adhesión a hidrocarburos utilizando dos solventes orgánicos (xileno y cloroformo). En los estudios de adhesión, las levaduras se pusieron en contacto con soluciones de biopolímeros y luego se enfrentaron a diferentes células (fibroblastos humanos y de rata y células epiteliales Hep-2). La cuantificación se realizó por microscopía óptica. Resultados: Se observó una disminución del 44% de la hidrofobicidad en presencia de QAPM y del 82%, con NaAL, o del 30% con QAPM y 19% con NaAL, cuando los solventes orgánicos empleados fueron cloroformo o xileno, respectivamente. La adhesión de C. albicans a células epiteliales y fibroblastos humanos disminuyó significativamente con ambos biopolímeros. En el caso de los fibroblastos de encía de rata, sólo se observó una disminución con NaAL. En ninguno de los experimentos se observaron diferencias significativas en asociación al tipo de fibroblasto empleado. Conclusiones: Los biopolímeros resultaron efectivos en la reducción de la hidrofobicidad y la adhesión de C. albicans a células, las cuales son importantes factores de virulencia relacionados con la colonización de los tejidos blandos del hospedador o superficies acrílicas presentes en el sistema estomatognático.Medicina oral, patologia oral y cirugia bucal 01/2006; · 1.10 Impact Factor