Antigenic analysis of coccidioidin and spherulin determined by two-dimensional immunoelectrophoresis

Huppert, M.; Spratt, N S; Vukovich, Katherine; Sun, Steven; Rice, E H

doi:10.1128/iai.20.2.541-551.1978

Cited by 55 publications

(56 citation statements)

References 29 publications

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“…Comparison of the intermediate-gel IEP plates in Fig. 4C and D demonstrated that the rabbit antiserum raised against the OG-SF of SOW contained antibodies against antigens 2 and CS of the reference system (4,14,19).…”

Section: Resultsmentioning

confidence: 94%

“…56,1988 on August 5, 2020 by guest http://iai.asm.org/ Downloaded from was analyzed in our earlier investigations (4, 5) by twodimensional immunoelectrophoresis (IEP). The coccidioidin (CDN)-anti-CDN reference system used for identification of specific antigens was originally developed by Huppert and co-workers (14,19) and has been reproduced in our laboratory with anticoccidioidin obtained from newly immunized burros. The procedure used for IEP examination in this study involved preparation of an intermediate gel (22) composed of 1 part rabbit serum (either preimmune serum or antiserum raised against OG-SF) to 1 part 2% agarose.…”

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confidence: 99%

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Isolation and morphology of an immunoreactive outer wall fraction produced by spherules of Coccidioides immitis

et al. 1988

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A previously undescribed, immunoreactive, membranous spherule outer wall (SOW) fraction produced by Coccidioides immitis (strains 634 and 735) grown in culture was isolated. Both this fraction and intact spherules were reactive with sera from coccidioidomycosis patients, as demonstrated by immunofluorescence microscopy. The serological activity of SOW was also demonstrated by its reactivity with human anti-C. immitis tube precipitin in a standardized immunodiffusion assay. Extraction of SOW with the nonionic detergent N-octyl-P-D-glucopyranoside (OG) permitted the isolation of an OG-soluble fraction which was reactive in the immunodiffusion assay. Rabbit antisera raised against the OG-soluble fraction were used in immunofluorescence and immunoelectron-microscopic studies of the parasitic cycle to confirm that the immunoreactive components of the solubilized fraction of SOW were associated with the inner and outer layers of the spherule wall as well as with distinct cytoplasmic organelles observed in thin sections of spherules. The immunoreactivity of SOW with sera from patients suggested that infected individuals are exposed to this surface wall material isolated from in vitro-grown spherules.decanted onto sterile Whatman no. 1 filter paper and subjected to aspirator-vacuum filtration. The filtrate was centrifuged (10,000 x g, 15 min, 4°C), and the supernatant was 2686 on August 5, 2020 by guest http://iai.asm.org/ Downloaded from 479-485.

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Section: Resultsmentioning

confidence: 94%

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confidence: 99%

Isolation and morphology of an immunoreactive outer wall fraction produced by spherules of Coccidioides immitis

et al. 1988

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“…Quantitative and q been reported in stu INFECT. IMMUN. etween Histoplasma and mycelia and yeast (or spherule) phases of these etween Histoplasma and three dimorphic fungi (10,18,19). Recently, trast to blastomycin KCB-Huppert et al (19) demonstrated 26 protein-1) did not elicit cross-reac-containing bands in coccidioidin (XV) using two--reactive subjects.…”

Section: Resultsmentioning

confidence: 99%

“…Antigens. Antigens from the saprophytic and parasitic (tissue) phases of C. immitis, H. capsulatum, and B. dermatitidis differ qualitatively and quantitatively (10,18,19). For this reason, LT assays were performed with antigens derived from mycelial and yeast (or spherule) phases of the three fungi.…”

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Cross-Reactivity Between Antigens of Coccidioides immitis, Histoplasma capsulatum , and Blastomyces dermatitidis in Lymphocyte Transformation Assays

Cox

1979

Infect Immun

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The cross-reactivity of four Coccidioides antigens, three Histoplasma antigens, and two Blastomyces antigens were determined in lymphocyte transformation assays of 11 coccidioidin-reactive, histoplasmin-nonreactive subjects (group I), 13 coccidioidin-nonreactive, histoplasmin-reactive persons (group II), and 13 subjects who were skin test negative to both antigens (group III). Mycelial and yeast (or spherule)-phase antigens of the three fungi were included. Significant crossreactivity was obtained with both coccidioidins, spherulin, and the alkali-soluble, water-soluble cell wall antigen of C. immitis, to the extent that the responses of histoplasmin-reactive persons were not statistically different (P > 0.05) from those of coccidioidin-reactive persons. In contrast, optimal dilutions of Histoplasma mycelial and yeast-phase lysates effectively distinguished (P < 0.01) responses of histoplasminand coccidioidin-reactive persons. The alkali-soluble cell wall antigen of H. capsulatum showed extensive cross-reactivity at most concentrations and was markedly stimulatory to lymphocytes of skin test-negative persons. Blastomycin elicited significant cross-reactions in histoplasmin-sensitive subjects and to a lesser extent in coccidioidin-sensitive subjects. The alkali-soluble cell wall antigen cross-reacted in cultures of histoplasmin-reactive persons but not in those of coccidioidin-reactive persons. All antigens effectively distinguished (P < 0.001) homologous responses of skin test-positive persons (groups I and II) from those of skin test-negative persons (group III). The extensive cross-reactivity in lymphocyte transformation assays in the absence of cross-reactivity in skin tests suggests that these two immune responses may be mediated by different T lymphocyte populations, may be elicited by different antigenic components, or both.

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“…MATERIALS AND METHODS Antigen preparations. Coccidioidin was prepared as described previously (9). Briefly, 12 isolates of C. immitis were grown separately in 2% glucose-1% yeast extract dialysate broth on a rotary shaker at 30 to 330C.…”

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Common antigens among systemic disease fungi analyzed by two-dimensional immunoelectrophoresis

Huppert¹,

Adler²,

Rice³

et al. 1979

Infect Immun

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The interpretation of immunological results in systemic mycoses has been complicated by cross-reactions among specimens from patients with blastomycosis, coccidioidomycosis, and histoplasmosis. The fungal preparations used in these tests evidently contained one or more antigens in common. Two-dimensional immunoelectrophoresis techniques were used to determine the number of antigens contained in several soluble extracts from Blastomyces dermatitidis and Histoplasma capsulatum that were common with those demonstrable in a coccidiodin-anticoccidioidin reference system. A total of 12 and 10 common antigens were found in preparations from B. dermatitidis and H. capsulatum, respectively. In addition, the crude preparations from each species of fungus exhibited some qualitative and quantitative differences in composition of antigens. Use of two-dimensional immunoelectrophoresis for standardization of fungus extracts, for monitoring separation of species-specific antigens, and for testing humoral antibody response should add further refinement to correlations with clinical disease.

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Antigenic analysis of coccidioidin and spherulin determined by two-dimensional immunoelectrophoresis

Cited by 55 publications

References 29 publications

Isolation and morphology of an immunoreactive outer wall fraction produced by spherules of Coccidioides immitis

Isolation and morphology of an immunoreactive outer wall fraction produced by spherules of Coccidioides immitis

Cross-Reactivity Between Antigens of Coccidioides immitis, Histoplasma capsulatum , and Blastomyces dermatitidis in Lymphocyte Transformation Assays

Common antigens among systemic disease fungi analyzed by two-dimensional immunoelectrophoresis

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