scholarly journals Fluconazole Alters the Polysaccharide Capsule of Cryptococcus gattii and Leads to Distinct Behaviors in Murine Cryptococcosis

PLoS ONE ◽  
2014 ◽  
Vol 9 (11) ◽  
pp. e112669 ◽  
Author(s):  
Julliana Ribeiro Alves Santos ◽  
Rodrigo Assunção Holanda ◽  
Susana Frases ◽  
Mayara Bravim ◽  
Glauber de S. Araujo ◽  
...  
mSphere ◽  
2018 ◽  
Vol 3 (2) ◽  
Author(s):  
Julia Catarina Vieira Reuwsaat ◽  
Heryk Motta ◽  
Ane Wichine Acosta Garcia ◽  
Carolina Bettker Vasconcelos ◽  
Bárbara Machado Marques ◽  
...  

ABSTRACTThe yeast-like pathogenCryptococcus gattiiis an etiological agent of cryptococcosis. The major cryptococcal virulence factor is the polysaccharide capsule, which is composed of glucuronoxylomannan (GXM), galactoxylomannan (GalXM), and mannoproteins (MPs). The GXM and GalXM polysaccharides have been extensively characterized; however, there is little information about the role of mannoproteins in capsule assembly and their participation in yeast pathogenicity. The present study characterized the function of a predicted mannoprotein fromC. gattii, designated Krp1. Loss-of-function and gain-of-function mutants were generated, and phenotypes associated with the capsular architecture were evaluated. The null mutant cells were more sensitive to a cell wall stressor that disrupts beta-glucan synthesis. Also, these cells displayed increased GXM release to the culture supernatant than the wild-type strain did. The loss of Krp1 influenced cell-associated cryptococcal polysaccharide thickness and phagocytosis by J774.A1 macrophages in the early hours of interaction, but no difference in virulence in a murine model of cryptococcosis was observed. In addition, recombinant Krp1 was antigenic and differentially recognized by serum from an individual with cryptococcosis, but not with serum from an individual with candidiasis. Taken together, these results indicate thatC. gattiiKrp1 is important for the cell wall structure, thereby influencing capsule assembly, but is not essential for virulencein vivo.IMPORTANCECryptococcus gattiihas the ability to escape from the host’s immune system through poorly understood mechanisms and can lead to the death of healthy individuals. The role of mannoproteins inC. gattiipathogenicity is not completely understood. The present work characterized a protein, Kpr1, that is essential for the maintenance ofC. gattiimain virulence factor, the polysaccharide capsule. Our data contribute to the understanding of the role of Kpr1 in capsule structuring, mainly by modulating the distribution of glucans inC. gattiicell wall.


2019 ◽  
Vol 9 (4) ◽  
pp. 330-340
Author(s):  
Mitradev Pattoo ◽  
Vuyokazi Belewa ◽  
Benesh Munilal Somai

Background:In both the developed and developing world, the mortality rates of people afflicted with cryptococcosis are unacceptably high despite the availability of antifungal therapy. The disease is caused by Cryptococcus neoformans (predominantly in immunocompromised individuals) and by Cryptococcus gattii. Globally the disease is estimated to cause around 600,000 deaths annually. Antifungal therapy is available, but in the developing world, may be unaffordable to many people, there is an increasing threat of resistance to the available drugs and our repertoire of antifungal drugs is very limited. Consequently, more research has been focusing on the use of medicinal plants as therapeutic agents. The originality of the current study is that although Tulbaghia violacea is a well-documented medicinal plant, the chemical composition of aqueous extracts and their antifungal potential against pathogenic yeasts are unknown. This is the first study that evaluates the chemical constituents of aqueous T. violacea root, leaf, rhizome and tuber extracts and their corresponding antifungal activities against C. neoformans and C. gattii.Objectives:The study aimed to investigate the phytochemical composition and antifungal potential of Tulbaghia violacea root, leaf, rhizome and tuber extracts against Cryptococcus neoformans and Cryptococcus gattii.Methods:Roots, leaves, rhizomes and tubers were extracted with water only for 48 h at room temperature with continuous shaking. Extracts were filter sterilized, freeze-dried and, chemically analyzed for saponin, flavonol, phenolic and tannin content. Chemical constituents of each extract were also identified by GC-MS analysis. The Minimum Inhibitory Concentration (MIC) of suitably diluted extracts of each plant part were also performed against C. neoformans and C. gattii, yeast pathogens commonly associated with HIV/AIDS sufferers.Results:Phytochemical analysis showed different concentrations of saponins (between 1023 and 2896.73 µg/ml), phenolics (between 16.48 and 51.58 µg/ml) and tannins (between 122.30 and 543.07 µg/ml) present in the different extracts. No flavonols were detected. GC-MS analysis identified a complex mixture of phytochemicals composed predominantly of sulphide, pyran, furan and ketone containing compounds to be present in the different plant parts. All extracts were dominated by the presence of 4 H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, a pyran known to have antifungal properties. Although the root, leaf, rhizome and tuber extracts exhibited antifungal activities against both fungi, the rhizome and tuber extract were found to possess the lowest MIC’s of 1.25 mg/ml and 2.5 mg/ml against Cryptococcus neoformans and Cryptococcus gattii respectively.Conclusion:T. violacea extracts have a complex constituent of phytochemicals and each plant part exhibited a strong antifungal activity against C. neoformans and C. gattii. The rhizome and tuber extracts showed the highest antifungal activity against C. neoformans and C. gattii respectively. Thus, T. violacea aqueous extracts are strong candidates for further development into an antifungal chemotherapeutic agent.


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