Preliminary characterization of exosome-like vesicles isolated from the supernatant of Paracoccidioides brasiliensis cultures (2006)
- Authors:
- Autor USP: TABORDA, CARLOS PELLESCHI - ICB
- Unidade: ICB
- Assunto: MICROBIOLOGIA
- Language: Inglês
- Abstract: Paracoccidioides brasiliensis is the dimorphic fungus responsible for paracoccidioidomycosis (PCM) in humans. In some organisms, vesicular structures, known as exosomes, are derived from the exocytic fusion of multivesicular endosomes with the cell surface. Exosomes have been recognized as important structures related with virulence of microorganisms and modulation of the host's immunity. Previous studies have supported the view that exosomes perform an acellular mode of communication and intercellular transfer of effector molecules. Exosomes have originally been isolated from several types of mammalian cells and more recently from pathogenic microorganisms like Trypanosoma cruzi (Torrecilhas et al., submitted). Here we describe the isolation of exosome-like vesicular structures for the first time in pathogenic dimorphic fungi. Supernatant fluids from P. brasiliensis B-339 yeast cells cultivated in minimum medium enriched with fetal calf serum were filtered and concentrated in an Amicon system. Half of the material was ultracentrifuged (100,000g) and analyzed by electron microscopy, where we could observe membranous vesicles sizing from 20 to 100 nm, not seen in preparations with medium alone. The remaining material was fractionated by gel-filtration chromatography. The eluted fractions were analyzed by ELISA with a reactive serum from a PCM patient. We observed that the highmolecular- weight peak fractions corresponding to the elution of vesicular structures werereactive with the PCM serum, suggesting the presence of P. brasiliensis antigens. Controls with sera from healthy individuals and fractionated medium alone were not reactive. In order to test their immunological relevance, P. brasiliensis ultracentrifuged vesicles were further fractionated in a sucrose gradient. We observed that gradient fractions 7 and 9 were the most active in the induction of nitric oxide by J774 cultured macrophages, while induction of IL-10 and IL-12 was negligible in our experimental conditions. The finding of extracellular vesicles that potentially carry immunologically active components opens new perspectives to the study of the host-parasite relationship in PCM. We are currently performing a proteomic analysis of the vesicle fractions.
- Imprenta:
- Publisher: American Society for Microbiology
- Publisher place: Denver
- Date published: 2006
- Source:
- Título do periódico: Abstracts
- Conference titles: ASM Conference on Dimorphic Fungal Pathogens
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ABNT
MATSUO, A. L. et al. Preliminary characterization of exosome-like vesicles isolated from the supernatant of Paracoccidioides brasiliensis cultures. 2006, Anais.. Denver: American Society for Microbiology, 2006. . Acesso em: 30 abr. 2024. -
APA
Matsuo, A. L., Ganiko, L., Freymüller, E., Silva, C., Taborda, C. P., Almeida, I. C., & Puccia, R. (2006). Preliminary characterization of exosome-like vesicles isolated from the supernatant of Paracoccidioides brasiliensis cultures. In Abstracts. Denver: American Society for Microbiology. -
NLM
Matsuo AL, Ganiko L, Freymüller E, Silva C, Taborda CP, Almeida IC, Puccia R. Preliminary characterization of exosome-like vesicles isolated from the supernatant of Paracoccidioides brasiliensis cultures. Abstracts. 2006 ;[citado 2024 abr. 30 ] -
Vancouver
Matsuo AL, Ganiko L, Freymüller E, Silva C, Taborda CP, Almeida IC, Puccia R. Preliminary characterization of exosome-like vesicles isolated from the supernatant of Paracoccidioides brasiliensis cultures. Abstracts. 2006 ;[citado 2024 abr. 30 ] - Therapeutic DNA vaccine encoding peptide p10 against experimental paracoccidioidomycosis
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