Fluconazole Alters the Polysaccharide Capsule of Cryptococcus gattii and Leads to Distinct Behaviors in Murine Cryptococcosis
Figure 2
L27/01 developed reduced susceptibility to fluconazole with overexpression of the MDR1 gene, but the parental strain is more virulent than L27/01F in mice.
Pulsed field gel electrophoresis (PFGE) patterns of L27/01F and L27/01 and strains on 1% PFGE certified agarose (lanes 2 and 3, respectively). Lane 1 show PFGE Size Marker (SM), 0.225–2.2 Mb S. cerevisiae chromosomal DNA Saccharomyces cerevisiae used as size standard (A). MDR1 gene expression in L27/01 and L27/01F strains (B). Ergosterol levels of L27/01 and L27/01F strains (C). Urease and Laccase activities of L27/01 and L27/01F strains: one enzymatic unit was calculated per 105 and 108 cells of C. gattii for urease and laccase activity, respectively, after 24 hours (D). Eight mice per group were inoculated by the intratracheal route with 106 cells of L27/01 or L27/01F. Animals administered with PBS represent the non-infected (control) group. Animals were monitored daily. Survival curve (E). Weight variation (Score) (F). Each 1 (one) gram received or lost corresponds to 10 points added or subtracted, respectively. *The vertical dotted line indicates a significant difference from the appointed day. CFU/g of lungs (G) from animals infected with 1×106 cells of L27/01 or L27/01F strains treated i.p. with fluconazole at 10 mg/kg/day (n = 6). NI: not infected group. NT: non treated group. Black bars refer to L27/01 (▪) and grey bars refer to L27/01F (▴). Data represent the mean of three independent experiments in triplicate. *P<0.05 was considered to be significant.