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Published on: April 15, 2019
Differences in cell morphometry, cell wall topography and gp70 expression correlate with the virulence of Sporothrix
Rafaela A Castro1, Paula H Kubitschek-Barreira, Pedro A C Teixeira
1Laboratório de Micologia Celular e Proteômica, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro UERJ, Rio de Janeiro, Brazil.
Abstract:
Sporotrichosis is a chronic infectious disease affecting both humans and animals. For many years, this subcutaneous mycosis had been attributed to a single etiological agent; however, it is now known that this taxon consists of a complex of at least four pathogenic species, including Sporothrix schenckii and Sporothrix brasiliensis. Gp70 was previously shown to be an important antigen and adhesin expressed on the fungal cell surface and may have a key role in immunomodulation and host response. The aim of this work was to study the virulence, morphometry, cell surface topology and gp70 expression of clinical isolates of S. brasiliensis compared with two reference strains of S. schenckii. Several clinical isolates related to severe human cases or associated with the Brazilian zoonotic outbreak of sporotrichosis were genotyped and clustered as S. brasiliensis. Interestingly, in a murine subcutaneous model of sporotrichosis, these isolates showed a higher virulence profile compared with S. schenckii. A single S. brasiliensis isolate from an HIV-positive patient not only showed lower virulence but also presented differences in cell morphometry, cell wall topography and abundant gp70 expression compared with the virulent isolates. In contrast, the highly virulent S. brasiliensis isolates showed reduced levels of cell wall gp70. These observations were confirmed by the topographical location of the gp70 antigen using immunoelectromicroscopy in both species. In addition, the gp70 molecule was sequenced and identified using mass spectrometry, and the sequenced peptides were aligned into predicted proteins using Blastp with the S. schenckii and S. brasiliensis genomes.
Insights
Sporothrix brasiliensis exhibits higher virulence than Sporothrix schenckii in sporotrichosis models. Virulence correlates inversely with cell wall gp70 expression in S. brasiliensis.
Area of Science:
- Mycology
- Infectious Diseases
- Immunology
Background:
- Sporotrichosis is a chronic fungal infection caused by Sporothrix species.
- Sporothrix schenckii and Sporothrix brasiliensis are key pathogenic species.
- Gp70 is a fungal surface antigen involved in host-pathogen interactions.
Purpose of the Study:
- Compare virulence, morphometry, and gp70 expression of clinical S. brasiliensis isolates with S. schenckii.
- Investigate the role of gp70 in S. brasiliensis virulence.
- Characterize gp70 in different clinical isolates.
Main Methods:
- Genotyping of clinical isolates.
- Murine subcutaneous model of sporotrichosis.
- Immunoelectron microscopy for gp70 localization.
- Mass spectrometry and peptide sequencing of gp70.
- Bioinformatic analysis (Blastp).
Main Results:
- Clinical S. brasiliensis isolates showed higher virulence than S. schenckii in mice.
- A less virulent S. brasiliensis isolate (from an HIV+ patient) displayed altered morphometry, cell wall topography, and abundant gp70.
- Highly virulent S. brasiliensis isolates had reduced cell wall gp70 expression.
- Gp70 topographical location varied between virulent and less virulent isolates.
Conclusions:
- S. brasiliensis is generally more virulent than S. schenckii.
- Cell wall gp70 expression is inversely correlated with S. brasiliensis virulence.
- Gp70's location and abundance are key factors in sporotrichosis pathogenesis.
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