Pathogenic Delivery: The Biological Roles of Cryptococcal Extracellular Vesicles

Oliveira, Haroldo César de; Castelli, Rafael F; Reis, Flavia C G; Rizzo, Juliana; Rodrigues, Márcio L.

doi:10.3390/pathogens9090754

Cited by 13 publications

(18 citation statements)

References 87 publications

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“…The presence of these moonlighting proteins in vesicles produced by other fungi has already been demonstrated for Histoplasma capsulatum [ 119 ], Cryptococcus neoformans [ 118 ], and Saccharomyces cerevisiae [ 120 ]. These moonlighting proteins are usually present in increased levels in the fungus cell wall during interaction with host cells, suggesting they may be involved in host–parasite interactions and virulence [ 95 , 116 , 121 , 122 , 123 ]. Some of these proteins have already been demonstrated to be crucial vaccine candidates in pathogenic fungi (e.g., glyceraldehyde-3-phosphate dehydrogenase, enolase, 14-3-3 protein, and fructose-1,6-bisphosphate aldolase), since they are highly expressed and have low identity with homolog proteins in the human host [ 124 , 125 ].…”

Section: Discussionmentioning

confidence: 99%

Immunoproteomic Analysis Reveals Novel Candidate Antigens for the Diagnosis of Paracoccidioidomycosis Due to Paracoccidioides lutzii

Rodrigues

Kubitschek-Barreira

Pinheiro

et al. 2020

JoF

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Paracoccidioidomycosis (PCM) is a life-threatening systemic infection caused by the fungal pathogen Paracoccidioides brasiliensis and related species. Whole-genome sequencing and stage-specific proteomic analysis of Paracoccidioides offer the opportunity to profile humoral immune responses against P. lutzii and P. brasiliensis s. str. infection using innovative screening approaches. Here, an immunoproteomic approach was used to identify PCM-associated antigens that elicit immune responses by combining 2-D electrophoresis of P. lutzii and P. brasiliensis proteomes, immunological detection using a gold-standard serum, and mass spectrometry analysis. A total of 16 and 25 highly immunoreactive proteins were identified in P. lutzii and P. brasiliensis, respectively, and 29 were shown to be the novel antigens for Paracoccidioides species, including seven uncharacterized proteins. Among the panel of proteins identified, most are involved in metabolic pathways, carbon metabolism, and biosynthesis of secondary metabolites in both immunoproteomes. Remarkably, six isoforms of the surface-associated enolase in the range of 54 kDa were identified as the major antigens in human PCM due to P. lutzii. These novel immunoproteomes of Paracoccidioides will be employed to develop a sensitive and affordable point-of-care diagnostic assay and an effective vaccine to identify infected hosts and prevent infection and development of human PCM. These findings provide a unique opportunity for the refinement of diagnostic tools of this important neglected systemic mycosis, which is usually associated with poverty.

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Section: Discussionmentioning

confidence: 99%

Immunoproteomic Analysis Reveals Novel Candidate Antigens for the Diagnosis of Paracoccidioidomycosis Due to Paracoccidioides lutzii

Rodrigues

Kubitschek-Barreira

Pinheiro

et al. 2020

JoF

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“…coming from inside the mother cell during budding. Besides being essential for budding, the presence of these SRs could be used, for example, as outlets for extracellular vesicles, whose mechanisms of passage through the cell wall are still unclear (de Oliveira et al, 2020). Our results show extracellular vesicles in the periplasmic space surrounding the SRs (Figure 5D).…”

Section: Discussionmentioning

confidence: 67%

“…We conjecture that the formation of these specialized regions, originated from the opening of mother cell walls and showing separated cell wall layers with thick tips delimiting the region, is related to a mechanical protruding force coming from inside the mother cell during budding. Besides being essential for budding, the presence of these SRs could be used, for example, as outlets for extracellular vesicles, whose mechanisms of passage through the cell wall are still unclear ( de Oliveira et al, 2020 ). Our results show extracellular vesicles in the periplasmic space surrounding the SRs ( Figure 5D ).…”

Section: Discussionmentioning

confidence: 99%

“…After production, Cryptococcus spp. PS can be either secreted to the extracellular milieu through vesicles ( Rodrigues et al, 2007 ; de Oliveira et al, 2020 ) or transported to the cell wall, where it forms the physical structure of the capsule in situ . Depending on its fate, PS acquires different physicochemical and rheological properties ( Pontes and Frases, 2015 ; Araújo et al, 2019 ; Zaragoza, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

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Ultrastructural Study of Cryptococcus neoformans Surface During Budding Events

et al. 2021

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Cryptococcus neoformans is a fungal pathogen that causes life-threatening infections in immunocompromised individuals. It is surrounded by three concentric structures that separate the cell from the extracellular space: the plasma membrane, the cell wall and the polysaccharide (PS) capsule. Although several studies have revealed the chemical composition of these structures, little is known about their ultrastructural organization and remodeling during C. neoformans budding events. Here, by combining the latest and most accurate light and electron microscopy techniques, we describe the morphological remodeling that occurs among the capsule, cell wall and plasma membrane during budding in C. neoformans. Our results show that the cell wall deforms to generate a specialized region at one of the cell’s poles. This region subsequently begins to break into layers that are slightly separated from each other and with thick tips. We also observe a reorganization of the capsular PS around the specialized regions. While daughter cells present their PS fibers aligned in the direction of budding, mother cells show a similar pattern but in the opposite direction. Also, daughter cells form multilamellar membrane structures covering the continuous opening between both cells. Together, our findings provide compelling ultrastructural evidence for C. neoformans surface remodeling during budding, which may have important implications for future studies exploring these remodeled specialized regions as drug-targets against cryptococcosis.

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“…EVs are likely to participate in cell wall remodeling ( Longo et al., 2014 ; Nimrichter et al., 2016 ; Zhao et al., 2019 ; reviewed in Rizzo et al., 2020 ) as well as in the synthesis of Candida biofilms ( Zarnowski et al., 2018 ; Zarnowski et al., 2021 ). They participate in the synthesis of the main virulence factor in Cryptococcus , which is the high molecular weight glucuronoxylomannose component of the capsule ( Rodrigues et al., 2008 ; Reis et al., 2019 ; reviewed in de Oliveira et al, 2020 ; Rizzo et al., 2020 ). The presence of peptides has been recently reported in Cryptococcus EVs, and one of them proved to protect against infection in G. mellonella ( Reis et al., 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Extracellular Vesicles From Paracoccidioides brasiliensis Can Induce the Expression of Fungal Virulence Traits In Vitro and Enhance Infection in Mice

Octaviano

Abrantes

Puccia

2022

Front. Cell. Infect. Microbiol.

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Extracellular vesicles (EVs) are cellular components involved in cargo delivery to the extracellular environment, including the fungal cell wall. Their importance in cell–cell communication, cell wall remodeling, and fungal virulence is starting to be better explored. In the human pathogenic Paracoccidioides spp., our group has pioneered the description of the EV secretome, carbohydrate cargo, surface oligosaccharide ligands, lipid, and RNA content. Presently, we studied the role of fungal EVs in the context of the virulent/attenuated model of the P. brasiliensis Pb18 isolate, which consists of variants transiently displaying higher (vPb18) or attenuated (aPb18) virulence capacity. In this model, the virulence traits can be recovered through passages of aPb18 in mice. Here, we have been able to revert the aPb18 sensitivity to growth under oxidative and nitrosative stress upon previous co-incubation with vEVs from virulent vPb18. That was probably due to the expression of antioxidant molecules, considering that we observed increased gene expression of the alternative oxidase AOX and peroxiredoxins HYR1 and PRX1, in addition to higher catalase activity. We showed that aEVs from aPb18 stimulated macrophages of the RAW 264.7 and bone marrow-derived types to express high levels of inflammatory mediators, specifically, TNF-α, IL-6, MCP-1, and NO. In our experimental conditions, subcutaneous treatment with EVs (three doses, 7-day intervals) before vPb18 challenge exacerbated murine PCM, as concluded by higher colony-forming units in the lungs after 30 days of infection and histopathology analysis. That effect was largely pronounced after treatment with aEVs, probably because the lung TNF-α, IFN-γ, IL-6, and MCP-1 concentrations were specially increased in aEV-treated when compared with vEV-treated mice. Our present studies were performed with EVs isolated from yeast cell washes of confluent cultures in Ham’s F-12 defined medium. Under these conditions, vEVs and aEVs have similar sizes but probably distinct cargo, considering that vEVs tended to aggregate upon storage at 4°C and −20°C. Additionally, aEVs have decreased amounts of carbohydrate and protein. Our work brings important contribution to the understanding of the role of fungal EVs in cell–cell communication and on the effect of EVs in fungal infection, which clearly depends on the experimental conditions because EVs are complex and dynamic structures.

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Pathogenic Delivery: The Biological Roles of Cryptococcal Extracellular Vesicles

Cited by 13 publications

References 87 publications

Immunoproteomic Analysis Reveals Novel Candidate Antigens for the Diagnosis of Paracoccidioidomycosis Due to Paracoccidioides lutzii

Immunoproteomic Analysis Reveals Novel Candidate Antigens for the Diagnosis of Paracoccidioidomycosis Due to Paracoccidioides lutzii

Ultrastructural Study of Cryptococcus neoformans Surface During Budding Events

Extracellular Vesicles From Paracoccidioides brasiliensis Can Induce the Expression of Fungal Virulence Traits In Vitro and Enhance Infection in Mice

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