Antigenic, chemical, and structural properties of cell walls of Histoplasma capsulatum yeast-form chemotypes 1 and 2 after serial enzymatic hydrolysis.

Cell walls of Histoplasma capsulatum yeast-form chemotypes 1 (chem 1) and 2 (chem 2) treated sequentially with several polysaccharolytic enzymes and Pronase yielded soluble, nondialyzable polysaccharides at each step, which were analyzed for monosaccharides, protein composition, and serological activity. Polysaccharide recovered after digestion of chem 1 walls with beta(1-->3)-glucanase contained glucose > mannose > glucosamine > galactose. This fraction (chem 1 betaG(1)) was analyzed by polyacrylamide gel electrophoresis and contained a component having an apparent molecular weight of 120,000. The chem 1 betaG(1) fraction was reactive in immunodiffusion (ID), producing an immune precipitate not identical to the H and M factors of histoplasmin. In a side-by-side ID comparison with extracts of chem 2, the chem 1 betaG(1) antigen contained an additional determinant not found in chem 2 extracts when tested with goat antiserum to H. capsulatum. Therefore, the chem 1 antigen gave preliminary ID evidence of antigenic group specificity. A chemical difference observed was the absence of glucosamine from chem 2 polysaccharide. In complement fixation (CF) tests, 9 of 17 sera from human histoplasmosis patients reacted with chem 1 betaG(1), but some cross-reactivity with sera of patients with other systemic mycoses occurred. The immunoelectrophoretic patterns of chem 1 wall-derived polysaccharides showed a marked shift in mobility after Pronase digestion, implying the presence of covalent peptides. The ultrastructural appearance and serological activity of intact walls and enzyme-resistant mural cores were also studied. The surface of the mural cores of both chemotypes was perforated and frayed. In shadow-cast preparations both fibrillar and globular areas persisted in the mural cores. The CF end point serum dilutions showed an increase after alpha- and beta-glucanase extractions of chem 2 walls and fourfold reduction after Pronase digestion. The mural cores of both chemotypes were still reactive in CF tests and retained some ability to bind fluorescent antibody. The chem 1 mural core reacted with specific fluorescein-labeled H. capsulatum antiglobulins produced by adsorption with Blastomyces dermatitidis, thus indicating at least partial retention of H. capsulatum-specific factors. The presence of galactose, mannose, and glucose was detected in the mural cores as well as enriched levels of amino sugar, despite exposure to chitinase.

Duke Scholars

Author Sara Elizabeth Miller Pathology