Interaction of murine macrophage-membrane proteins with components of the pathogenic fungus Histoplasma capsulatum
Por:
Taylor M.L., Duarte-Escalante E., Reyes-Montes M.R., Elizondo N., Maldonado G., Zenteno E.
Publicada:
1 ene 1998
Resumen:
The interaction of macrophage-membrane proteins and histoplasmin, a crude antigen of the pathogenic fungus Histoplasma capsulatum, was studied using murine peritoneal macrophages. Membrane proteins were purified via membrane attachment to polycationic beads and solubilized in Tris- HCl/SDS/DTT/glycerol for protein extraction; afterwards they were adsorbed or not with H. capsulatum yeast or lectin binding-enriched by affinity chromatography. Membrane proteins and histoplasmin interactions were detected by ELISA and immunoblotting assays using anti-H. capsulatum human or mouse serum and biotinylated goat anti-human or anti-mouse IgG/streptavidin- peroxidase system to reveal the interaction. Results indicate that macrophage-membrane proteins and histoplasmin components interact in a dose- dependent reaction, and adsorption of macrophage-membrane proteins by yeast cells induces a critical decrease in the interaction. Macrophage-membrane glycoproteins with terminal D-galactosyl residues, purified by chromatography with Abrus precatorius lectin, bound to histoplasmin; and two bands of 68 kD and 180 kD of transferred membrane protein samples interacted with histoplasmin components, as revealed by immunoblot assays. Specificity for ?-galactoside residues on the macrophage-membrane was confirmed by galactose inhibition of the interaction between macrophage-membrane proteins and histoplasmin components, in competitive ELISA using sugars, as well as by enzymatic cleavage of the galactoside residues.
Filiaciones:
Taylor M.L.:
Dept. of Microbiol.-Parasitol., Faculty of Medicine, UNAM, Mexico
Lab. de Inmunología de Hongos, Depto. de Microbiol. y Parasitol., Ciudad Universitaria, México, D.F. 04510, Mexico
Duarte-Escalante E.:
Dept. of Microbiol.-Parasitol., Faculty of Medicine, UNAM, Mexico
Reyes-Montes M.R.:
Dept. of Microbiol.-Parasitol., Faculty of Medicine, UNAM, Mexico
Elizondo N.:
Dept. of Microbiol.-Parasitol., Faculty of Medicine, UNAM, Mexico
Maldonado G.:
Department of Bioquímica, Faculty of Medicine, UNAM, Mexico
Zenteno E.:
Department of Bioquímica, Faculty of Medicine, UNAM, Mexico
Department of Bioquímica, Inst. Nac. de Enferm. Respiratorias, Mexico
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