Utilização de métodos de análise no fracionamento de biomoléculas potencialmente imunossupressoras do fungo ambiental Trichoderma stromaticum

• Ano de defesa: 2011
• Autor(a) principal: Cintia Lima dos Santos
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Minas Gerais; UFMG
• Palavras-chave em Português: Imunossupressoras; Trichoderma stromaticum; Biomolécular; Engenharia nuclear
• Link de acesso: http://hdl.handle.net/1843/MBAM-8L8NKJ

Trichoderma stromaticum is used as biocontrol agent of the cacao witches broom pathogen Moniliophthora perniciosa. The collected data showed that T. stromaticum spores inhibit the defense mechanisms of innate and adaptative immunity of mice. This suggests that the spores have biologically active substances with immunosuppressor effects. The goal of this research was fractionated T. stromaticum fungi molecules that show immunosuppressor potential when functioning as phagocytes. T. stromaticum spores grown on potato dextrose agar at room temperature, for 7-10 days were collected and heat-treated at 100 ºC for 15 min (spores inactive) or not (active spores) to later processing and evaluation of the ultrastructure through Transmission Eletron Microscopy (TEM). The active spores were used to the yield of the gross extract and later fractioning through Fast Protein Liquid Chromatography (FPLC). The fractions resulting from the chromatography were used in assays of nitrite oxide by macrophages through the Griess Reaction colorimetric and immunogenicity of the spores and ExtTs. The results showed that inactive spore ultrastructure revealed intracellular disorder and an increase in the thickness of the wall. The fractioned of the crude extract generated a pools fraction called 2 e 4, which significantly inhibited NO yield by macrophages activated with IFN- + LPS. Small concentrations of spore T. stromaticum change significantly ultraestructrure of murine macrophages. Such changes suggest the possiblility of occurrence of the process of autophagy. The method of analysis allowed to select a fraction of T. stromaticum with immunosuppressor potencial that constitues target of great interest to future molecular characterization due to its possible therapeutical relevance. Until then, our study suggests that the use of T. stromaticum as a biopesticide in aerosol devices should be performed carefully to avoid health problems to plantation workers.