Proteases of Sporothrix schenckii: Cytopathological effects on a host-cell model |
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Authors: | Myrna Sabanero López Lérida L Flores Villavicencio Karla Soto Arredondo Gloria Barbosa Sabanero Julio César Villagómez-Castro Gustavo Cruz Jiménez Gerardo Sandoval Bernal Haydee Torres Guerrero |
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Institution: | 1. Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta S/N, Col. Noria Alta, Guanajuato, Guanajuato 36000, Mexico;2. Departamento de Ciencias Médicas, División de Ciencias de la Salud, Campus León, Universidad de Guanajuato, 20 de Enero 292, Col. Obregón, León, Guanajuato 37320, Mexico;3. Departamento de Farmacia, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta S/N, Col. Noria Alta, Guanajuato, Guanajuato 36000, Mexico;4. Laboratorio de Micología Básica, Unidad de Medicina Experimental, Facultad de Medicina, UNAM, Dr. Balmis No. 148, Col. Doctores, Del. Cuauhtémoc, Mexico, DF, Mexico |
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Abstract: | BackgroundSporotrichosis is a fungal infection caused by the Sporothrix schenckii complex. The adhesion of the fungus to the host tissue has been considered the key step in the colonization and invasion, but little is known about the early events in the host–parasite interaction.AimsTo evaluate the proteolytic activity of S. schenckii on epithelial cells.MethodsThe proteolytic system (at pH 5 and 7) was evaluated using azocoll and zymograms. The host–parasite interaction and epithelial cell response were also analyzed by examining the microfilament cytoskeleton using phalloidin-FITC and transmission electron microscopy. Finally, the metabolic activity was determined using an XTT assay.ResultsThe zymograms showed that S. schenckii yeast cells possess high intracellular and extracellular proteolytic activities (Mr ≥ 200, 116, 97, and 70 kDa) that are pH dependent and are inhibited by PMSF and E64, which act on serine and cysteine-type proteases. During the epithelial cell–protease interaction, the cells showed alterations in the microfilament distribution, as well as in the plasma membrane structure. Moreover, the metabolic activity of the epithelial cells decreased 60% without a protease inhibitor.ConclusionsOur data demonstrate the complexity of the cellular responses during the infection process. This process is somehow counteracted by the action of proteases inhibitors. Furthermore, the results provide critical information for understanding the nature of host–fungus interactions and for searching a new effective antifungal therapy, which includes protease inhibitors. |
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Keywords: | Proteases Epithelium Interactions Cytoskeleton Proteasas Epitelio Interacción Citoesqueleto |
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