Peptide antigen preparation was purified from cultured
Sporothrix schenckii mycelia by extraction with 0.1 N-HCl and picric acid precipitation. This preparation—tentatively termed sporothrix peptide antigen (SPA)—elicited a delayed-type hypersensitivity reaction in immunized guinea pigs. The minimum dose required to elicit a positive skin reaction was 5
μg. SPA also specifically inhibited the migration of peritoneal exudate cells in immunized guinea pigs. Chemical analyses showed that SPA contained above 90 percent protein and a minute quantity of polysaccharide and nucleic acid. In enzyme digestion studies, the antigenic activity of SPA was completely destroyed by treatment with pronase. Thus, the peptide moiety probably carried the entire antigenic activity. SPA showed a widely distributed pattern, in gel-filtration and electrophoresis studies. The estimated molecular weight was 5,000—800,000 daltons or more. SPA also was stable to heat and acid-alkali processing: heating at 120°C or treatment with 3N-HCl or 3N-NaOH resulted in no change in the antigenic activity. A clinical evaluation of SPA was carried out in 26 patients with sporotrichosis. SPA and conventional sporotrichin produced almost the same degree of skin reaction in most patients, but in only 4 cases, SPA alone showed a positive skin reaction. The specificity of SPA was, therefore, estimated to be almost the same as or slightly higher than that of the conventional sporotrichin. The high specificity, heat stability, high antigenic activity (per unit weight) and highly purified nature of SPA suggest that it may be effective for clinical skin testing and
in vitro examinations of cellular immunity such as migration inhibition or lymphoblast formation tests.
View full abstract