Excretion-secretion products and proteases from live Sporothrix schenckii yeast phase: immunological detection and cleavage of human IgG.
Antigenic preparations from Sporothrix schenckii usually involve materials from mixed cultures of yeast and mycelia presenting cross-reactions with other deep mycoses. We have standardized pure yeast phase with high viability of the cells suitable to obtain specific excretion-secretion products without somatic contaminations. These excretion-secretion products were highly immunogenic and did not produce noticeable cross-reactions in either double immunodiffusion or Western blot. The antigenic preparation consists mainly of proteins with molecular weights between 40 and 70 kDa, some of them with proteolytic activity in mild acidic conditions. We also observed cathepsin-like activity at two days of culture and chymotrypsin-like activity at four days of culture consistent with the change in concentration of different secreted proteins. The proteases were able to cleave different subclasses of human IgG suggesting a sequential production of antigens and molecules that could interact and interfere with the immune response of the host.
Da Rosa
Daniel
D
Departamento de Parasitología y Micología, Universidad de la República, Montevideo, Uruguay.
Gezuele
Elbio
E
Calegari
Luis
L
Goñi
Fernando
F
D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology; D000946:Antigens, Fungal / Q000096:biosynthesis*; D002403:Cathepsins / Q000096:biosynthesis*; D002918:Chymotrypsin / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000096:biosynthesis*; D006801:Humans; D015151:Immunoblotting; D005779:Immunodiffusion; D007074:Immunoglobulin G / Q000276:immunology*; D008970:Molecular Weight; D011817:Rabbits; D013175:Sporothrix / Q000378:metabolism*
D000974:Antibodies, Antinuclear; D000946:Antigens, Fungal; D005656:Fungal Proteins; D007074:Immunoglobulin G; C035356:SS-A antibodies; D002403:Cathepsins; D002918:Chymotrypsin