Comparative serological and cutaneous reactivity of candidal cytoplasmic proteins and mannan separated by affinity for concanavalin A

Abstract

Yeast-form Candida albicans cells were disrupted for 1.5 min in a Braun homogenizer and centrifuged at 100,000 X g. The supernatant was concentrated by ammonium sulfate precipitation and then dialyzed. The resulting material (650 mg), containing 81.2% protein and 11.5% carbohydrate, was subjected to affinity chromatography on concanavalin A (Con A) linked to agarose. A protein fraction was eluted from the column with buffer, and a fraction containing mannan was eluted with 0.2 M alpha-methyl mannoside. The candidal soluble proteins had 19 components which were resolvable by polyacrylamide gel electrophoresis. The material with affinity for Con A contained mannan and 17% complexed protein. Antigenic differences between the soluble proteins and the mannan-protein complex were shown by lines of intersection in immunodiffusion. The soluble proteins devoid of mannan reacted in immunoelectrophoresis with sera from infected rabbits and patients with chronic candidiasis. These same sera also reacted with a mannan-protein complex eluted from the Con A column with alpha-methyl mannoside. The comparative ability of candidal proteins and cell wall-derived mannan to elicit skin test reactions in guinea pigs sensitized by infection or with formaldehyde-killed yeast was studied. Candidal proteins at a 10-mug dose elicited positive reactions at 6 and 21 days after sensitization. The reactions persisted for 48 h and showed minimal tendency to an arthus response, which was marked when mannan-containing antigens were used. The antigenicity of cell wall-derived mannans and candidal soluble proteins devoid of mannan was compared in immunodiffusion tests of sera from 39 patients with neoplastic disease. Of these patients with documented candidiasis, 13 of 20 reacted to one or more mannan antigens, and 3 of 20 reacted to candidal soluble proteins. In contrast, of those patients who were uninfected or had superficial Candida spp. infections, 5 of 19 reacted to candidal soluble proteins, and 16 of 19 reacted to one or more mannan antigens.