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Fluconazole alters the polysaccharide capsule of Cryptococcus gattii and leads to distinct behaviors in murine Cryptococcosis.

Santos JR, Holanda RA, Frases S, Bravim M, Araujo Gde S, Santos PC, Costa MC, Ribeiro MJ, Ferreira GF, Baltazar LM, Miranda AS, Oliveira DB, Santos CM, Fontes AC, Gouveia LF, Resende-Stoianoff MA, Abrahão JS, Teixeira AL, Paixão TA, Souza DG, Santos DA - PLoS ONE (2014)

Bottom Line: L27/01F presented lower antiphagocytic properties and reduced survival inside macrophages.The L27/01F did not affect the central nervous system, while the effect in brain caused by L27/01 strain began after only 12 hours.The behavioral alterations were affected by L27/01, but no effects were detected after infection with L27/01F.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Minas Gerais, Brazil.

ABSTRACT
Cryptococcus gattii is an emergent human pathogen. Fluconazole is commonly used for treatment of cryptococcosis, but the emergence of less susceptible strains to this azole is a global problem and also the data regarding fluconazole-resistant cryptococcosis are scarce. We evaluate the influence of fluconazole on murine cryptococcosis and whether this azole alters the polysaccharide (PS) from cryptococcal cells. L27/01 strain of C. gattii was cultivated in high fluconazole concentrations and developed decreased drug susceptibility. This phenotype was named L27/01F, that was less virulent than L27/01 in mice. The physical, structural and electrophoretic properties of the PS capsule of L27/01F were altered by fluconazole. L27/01F presented lower antiphagocytic properties and reduced survival inside macrophages. The L27/01F did not affect the central nervous system, while the effect in brain caused by L27/01 strain began after only 12 hours. Mice infected with L27/01F presented lower production of the pro-inflammatory cytokines, with increased cellular recruitment in the lungs and severe pulmonary disease. The behavioral alterations were affected by L27/01, but no effects were detected after infection with L27/01F. Our results suggest that stress to fluconazole alters the capsule of C. gattii and influences the clinical manifestations of cryptococcosis.

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L27/01F increases recruitment of host cells to the lungs, but decreases cytokine production.L27/01 and L27/01F strains elicit different inflammatory response in C57/BL6 mice. Mice (n = 8) were inoculated i.t. with 106 yeast cells of L27/01 or L27/01F strains. After 1, 7, 15 or 30 days post infection, the mice were euthanized and mononuclear leukocytes (A) and neutrophils (B) in bronchoalveolar lavage fluid (BALF) were quantified. Neutrophilic infiltrate in the lungs was also determined as MPO activity and expressed as relative number of neutrophils (C). Levels of CXCL1/KC (D), IFN-γ (E), TNF-α (F) and IL-10 (G) are expressed in pg per 100 mg of lung. White bars refer to not infected (NI) animals, black bars to animals infected with the L27/01 strain, and grey bars to animals infected with the L27/01F strain. The results are presented as the mean ± SEM. Each experiment was performed in triplicate. *P<0.05 was considered to be significant.
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pone-0112669-g006: L27/01F increases recruitment of host cells to the lungs, but decreases cytokine production.L27/01 and L27/01F strains elicit different inflammatory response in C57/BL6 mice. Mice (n = 8) were inoculated i.t. with 106 yeast cells of L27/01 or L27/01F strains. After 1, 7, 15 or 30 days post infection, the mice were euthanized and mononuclear leukocytes (A) and neutrophils (B) in bronchoalveolar lavage fluid (BALF) were quantified. Neutrophilic infiltrate in the lungs was also determined as MPO activity and expressed as relative number of neutrophils (C). Levels of CXCL1/KC (D), IFN-γ (E), TNF-α (F) and IL-10 (G) are expressed in pg per 100 mg of lung. White bars refer to not infected (NI) animals, black bars to animals infected with the L27/01 strain, and grey bars to animals infected with the L27/01F strain. The results are presented as the mean ± SEM. Each experiment was performed in triplicate. *P<0.05 was considered to be significant.

Mentions: Mononuclear leukocytes and neutrophils were more recruited to the lungs by the L27/01F strain than the L27/01 strain (p<0.05) (Figure 6A–B). The MPO activity in the lungs (Figure 6C) confirmed higher neutrophil influx with the F strain (p<0.05).


Fluconazole alters the polysaccharide capsule of Cryptococcus gattii and leads to distinct behaviors in murine Cryptococcosis.

Santos JR, Holanda RA, Frases S, Bravim M, Araujo Gde S, Santos PC, Costa MC, Ribeiro MJ, Ferreira GF, Baltazar LM, Miranda AS, Oliveira DB, Santos CM, Fontes AC, Gouveia LF, Resende-Stoianoff MA, Abrahão JS, Teixeira AL, Paixão TA, Souza DG, Santos DA - PLoS ONE (2014)

L27/01F increases recruitment of host cells to the lungs, but decreases cytokine production.L27/01 and L27/01F strains elicit different inflammatory response in C57/BL6 mice. Mice (n = 8) were inoculated i.t. with 106 yeast cells of L27/01 or L27/01F strains. After 1, 7, 15 or 30 days post infection, the mice were euthanized and mononuclear leukocytes (A) and neutrophils (B) in bronchoalveolar lavage fluid (BALF) were quantified. Neutrophilic infiltrate in the lungs was also determined as MPO activity and expressed as relative number of neutrophils (C). Levels of CXCL1/KC (D), IFN-γ (E), TNF-α (F) and IL-10 (G) are expressed in pg per 100 mg of lung. White bars refer to not infected (NI) animals, black bars to animals infected with the L27/01 strain, and grey bars to animals infected with the L27/01F strain. The results are presented as the mean ± SEM. Each experiment was performed in triplicate. *P<0.05 was considered to be significant.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231059&req=5

pone-0112669-g006: L27/01F increases recruitment of host cells to the lungs, but decreases cytokine production.L27/01 and L27/01F strains elicit different inflammatory response in C57/BL6 mice. Mice (n = 8) were inoculated i.t. with 106 yeast cells of L27/01 or L27/01F strains. After 1, 7, 15 or 30 days post infection, the mice were euthanized and mononuclear leukocytes (A) and neutrophils (B) in bronchoalveolar lavage fluid (BALF) were quantified. Neutrophilic infiltrate in the lungs was also determined as MPO activity and expressed as relative number of neutrophils (C). Levels of CXCL1/KC (D), IFN-γ (E), TNF-α (F) and IL-10 (G) are expressed in pg per 100 mg of lung. White bars refer to not infected (NI) animals, black bars to animals infected with the L27/01 strain, and grey bars to animals infected with the L27/01F strain. The results are presented as the mean ± SEM. Each experiment was performed in triplicate. *P<0.05 was considered to be significant.
Mentions: Mononuclear leukocytes and neutrophils were more recruited to the lungs by the L27/01F strain than the L27/01 strain (p<0.05) (Figure 6A–B). The MPO activity in the lungs (Figure 6C) confirmed higher neutrophil influx with the F strain (p<0.05).

Bottom Line: L27/01F presented lower antiphagocytic properties and reduced survival inside macrophages.The L27/01F did not affect the central nervous system, while the effect in brain caused by L27/01 strain began after only 12 hours.The behavioral alterations were affected by L27/01, but no effects were detected after infection with L27/01F.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Minas Gerais, Brazil.

ABSTRACT
Cryptococcus gattii is an emergent human pathogen. Fluconazole is commonly used for treatment of cryptococcosis, but the emergence of less susceptible strains to this azole is a global problem and also the data regarding fluconazole-resistant cryptococcosis are scarce. We evaluate the influence of fluconazole on murine cryptococcosis and whether this azole alters the polysaccharide (PS) from cryptococcal cells. L27/01 strain of C. gattii was cultivated in high fluconazole concentrations and developed decreased drug susceptibility. This phenotype was named L27/01F, that was less virulent than L27/01 in mice. The physical, structural and electrophoretic properties of the PS capsule of L27/01F were altered by fluconazole. L27/01F presented lower antiphagocytic properties and reduced survival inside macrophages. The L27/01F did not affect the central nervous system, while the effect in brain caused by L27/01 strain began after only 12 hours. Mice infected with L27/01F presented lower production of the pro-inflammatory cytokines, with increased cellular recruitment in the lungs and severe pulmonary disease. The behavioral alterations were affected by L27/01, but no effects were detected after infection with L27/01F. Our results suggest that stress to fluconazole alters the capsule of C. gattii and influences the clinical manifestations of cryptococcosis.

Show MeSH
Related in: MedlinePlus