Delayed hypersensitivity responses of experimental animals to histoplasmin from the yeast and mycelial phases of Histoplasma capsulatum

Abstract

Controlled yeast lysate (CYL) and controlled mycelial lysate (CML) histoplasmins were produced from Histoplasma capsulatum grown in a nutritionally lean, chemically defined medium. The lysates were assayed for skin-test activity in guinea pigs sensitized by infection with the homologous organism. In some studies, nonliving vaccine preparations were employed also. Inter-lot biological variation was animal, and 20 lots of the CYL reagent elicited strong dermal reactions with high specificity. Further, CYL reagents were nonreactive in guinea pigs infected with Coccidioides immitis, whereas the commercial Food and Drug Administration preparations cross-reacted to some degree. The CML histoplasnins were generally less reactive than the CYL preparations and exhibited somewhat more inter-lot variation in sensitivity and specificity. No correlation between potency and protein:polysaccharide ratios were observed with either reagent. An intradermal test with the CYL reagent did not induce significant changes in the complement-fixing titer of sensitized guinea pigs. Such changes in sensitized animals were elicited by a skin test with commercial histoplasmin.

Detection of Antibodies in Serum Specimens from Dogs with Blastomycosis with Lysate Antigens Prepared from Four <i>Blastomyces dermatitidis</i> Dog Isolates: Individual Antigens vs Antigen Combinations

“…Yeast phase lysate reagents (T-58, dog Tennessee, T-66, dog Tennessee, WI-R, dog Wisconsin and WI-J, dog Wisconsin) were prepared by a method similar to one that was previously used for the production of antigen from Histoplasma capsulatum [29,30] and modified in our laboratory for B. dermatitidis lysate antigen production [18]. The yeast phase cells were grown for 7 days at 37˚C in a chemically defined medium in an incubator shaker, harvested by centrifugation (700 × g; 5 min), followed by washing with distilled water, resuspended in distilled water and then allowed to lyse for 7 days at 37 C in water with shaking.…”

Comparison of enzyme immunoassay and immunodiffusion for the detection of canine blastomycosis

“…The above study led us to begin an evaluation of different methods for the preparation of B. dermatitidis antigens in an attempt to develop a reagent with sensitivity as well as specificity. A controlled yeast lysate antigen of H. capsulatum had previously been shown to be useful for the ELISA detection of anti-H. capsulatum antibodies [11] and also for the detection of delayed dermal hypersensitivity in animals and humans [18,19]. These results prompt-ed us to produce yeast phase lysate antigens from human, dog and soil isolates of B. dermatitidis by a method similar to the one used for the preparation of the H. capsulatum lysate reagent.…”

“…Yeast cell lysate antigens were prepared from human (K-Le) and dog (T-58) isolates of B. dermatitidis by a method similar to that used for the preparation of a controlled yeast phase lysate antigen from Histoplasma capsulatum [ 18,19]. The human isolate was obtained from Dr N. Goodman (University of Kentucky Medical Center, Lexington, KY) and the dog isolate from Dr A. M.…”

Comparison of Detection of Antibodies with Yeast Lysate Antigens Prepared from Two Isolates of <i>Blastomyces dermatitidis</i> by Two Different Methods: Sensitivity and Specificity Evaluations

“…Lysate Antigen Preparation Mycelial phase cultures of two B. dermatitidis isolates, B5896, a human isolate from Mountain Iron Minnesota and 597, a human isolate from Eagle River Wisconsin [28,29], were converted to yeast cells by culturing at 37˚C on brain heart infusion agar. Yeast phase lysate reagents were prepared by a method similar to one that was previously used for the production of antigen from Histoplasma capsulatum [30][31][32] and modified in our laboratory for B. dermatitidis lysate antigen production [19]. The yeast phase cells were grown for 7 days at 37˚C in a chemically defined medium in an incubator shaker, harvested by centrifugation (700 × g; 5 min), followed by washing with distilled water, resuspended in distilled water and then allowed to lyse for 1 or 7 days at 37˚C in water with shaking.…”