[Results of cultural vaginal smear studies as a proof of pathogenous Blastomyces. Laboratory diagnosis of Blastomyces infection of the female genitalia].
Explore the source record for details and available documents.
Bacteriological Techniques↗
SEARCH · Search PubMed
Results for “Blastomyces”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
[Comparative studies on the occurrence of blastomyces on healthy and sick skin. 2. Epidemiology of propagation of blastomyces on healthy and sick skin].
Explore the source record for details and available documents.
Blastomyces↗
[Comparative studies on the occurrence of blastomyces on healthy and diseased skin. 1. The occurrence of blastomyces on healthy and diseased skin].
Explore the source record for details and available documents.
Blastomyces↗
[Infusion liquids as the cause of blastomyces septicemia in childhood. On the development, management and prevention of iatrogenic blastomyces septicemia].
Explore the source record for details and available documents.
Amphotericin B↗
Purification and characterization of the major antigen WI-1 from Blastomyces dermatitidis yeasts and immunological comparison with A antigen.
The lack of well-defined antigens from Blastomyces dermatitidis has hampered the ability to reliably diagnose human infection and study the immunobiology of blastomycosis. We recently discovered a novel surface protein on B. dermatitidis yeasts, designated WI-1, and demonstrated it to be a key antigenic target of humoral and cellular responses during infection. In the present article, we purified and characterized WI-1 and compared it immunologically with the only Blastomyces antigen commercially available, A antigen. WI-1 was purified by high-performance liquid chromatography over a DEAE-cellulose column. It eluted from the column at a point on the salt gradient corresponding to 460 to 490 mM NaCl, reflecting its acidic pI of approximately equal to 5.2. Purified WI-1 had a molecular mass of 120 kDa and contained a large amount of cysteine (85 residues) and aromatic amino acids but undetectable carbohydrate. In contrast, A antigen had a molecular mass of 135 kDa and contained 37% carbohydrate. Immunological comparison of the two antigens showed that, when radiolabeled, WI-1 was more reactive with anti-Blastomyces antisera than A antigen but did not cross-react with anti-Histoplasma antisera. Proteinase digestion of WI-1 eliminated its recognition by anti-WI-1 and anti-Blastomyces antisera. Proteinase treatment of A antigen had no effect on its recognition by anti-Blastomyces or anti-Histoplasma antisera, but periodate treatment abolished recognition by anti-Histoplasma antisera, indicating that the cross-reactive determinant(s) of A antigen is displayed on the accompanying carbohydrate. In further studies, anti-WI-1 antiserum reacted with A antigen and, conversely, anti-A antiserum and monoclonal antibodies (MAbs) reacted with WI-1, indicating a shared determinant on the two antigens. A recombinant 25-amino-acid repeat, recently cloned from WI-1 and found to be the major target of antibody recognition of WI-1, reacted strongly with anti-A antiserum and MAbs. In MAb competition tests, MAbs specific for the 25-residue repeat abolished binding of anti-A antiserum to A antigen. In antigen inhibition tests, the recombinant repeat abolished binding of anti-A antiserum to A antigen. These results demonstrate that the repeat is the major site of antibody recognition of both WI-1 and A antigen and that the recombinant, nonglycosylated peptide could replace either native antigen in formatting better diagnostic tests for blastomycosis. Moreover, they suggest that producing fungal protein antigens as nonglycosylated peptides in a procaryotic expression system may circumvent problems of antigen cross-reactivity that are due to posttranslational modification.
Animals↗
Molecular taxonomy and epidemiology of Blastomyces and Histoplasma species.
Cladistic analysis of partial 26S rRNA sequences was used to estimate evolutionary distances among species and varieties of the dimorphic onygenalean genera Blastomyces, Coccidioides, Emmonsia, Histoplasma and Paracoccidioides. With the exception of Coccidioides, all genera were closely related, with about 5% base differences and even less (1-2%) between Blastomyces and Emmonsia. These data were supported by a teleomorph in the same genus Ajellomyces. In a phylogenic study of a wide range of ascomycete orders and families, Coccidioides immitis was found to be closest to Aphanoascus fulvescens and Chrysosporium keratinophilum, and to have relative distances to the remaining dimorphic genera (family Onygenaceae) similar to those of the dermatophytes (family Arthrodermataceae). The sequencing data were confirmed by genomic comparisons. All dimorphic genera had a nuclear DNA base composition in the same range of 46.6-47.3% G + C. The DNA melting curves of Blastomyces and Histoplasma strains showed irregularities that were ascribed to the presence of AT-rich stretches in satellite DNA rather than in mitochondrial DNA. Derivative profiles proved to be highly reproducible within regional populations and coincided with differences in clinical behaviour of each species. Blastomyces dermatitidis generated two kinds of curves, corresponding to the geographically distinct serotypes 1 and 2. The African type (serotype 2) was characterized by a classical sigmoidal melting curve similar to that for all strains of Coccidioides, Emmonsia and Paracoccidioides. In contrast, the American type (serotype 1) contained satellite DNA (27% G + C). A rRNA base difference of 1.5% was observed between geographical types, a value slightly higher than that noted between Histoplasma capsulatum and its variety farciminosum (0.9%). All three H. capsulatum varieties presented irregularities in their DNA melting curves. The molecular data support the recognition of two of them as agents of blastomycosis and the assignment of more than one species and two varieties to the genus Emmonsia.
Animals↗
In vivo and in vitro cell-mediated immune responses to a cell wall antigen of Blastomyces dermatitidis.
An alkali-soluble, water-soluble cell wall fraction of Blastomyces dermatitidis, designated B-ASWS, was evaluated as an antigen for detecting in vivo (skin tests) and in vitro migration inhibition factor (MIF) production and lymphocyte transformation (LT) responses in Blastomyces-infected guinea pigs. The biological activity of B-ASWS was compared with that of blastomycin KCB-26. The superiority of B-ASWS, in terms of its sensitivity and specificity, was evident in in vivo and in vitro assays. Skin tests responses were obtained in 21 of the 24 Blastomyces-infected guinea pigs, whereas only one of the 14 Histoplasma-infected guinea pigs were significantly greater than those obtained using cell populations from Histoplasma-infected or noninfected guinea pigs. The con-MIF and LT in peritoneal exudate cells and lymph node cells of homologuosly infected animals. In each biological system, the response of the Blastomyces-infected guinea pigs were significantly greater than those obtained using cell populations from Histoplasma-infected or non-infected guinea pigs. The contrasting efficacy of B-ASWS as compared with blastomycin KCB-26, suggests that the cell wall antigen will be a useful tool for detecting cell-mediated immune responses in blastomycosis.
Animals↗
Intracellular growth and phagocytosis of Blastomyces dermatitidis by monocyte-derived macrophages from previously infected and normal subjects.
Blastomyces dermatitidis evokes responses of human cellular immunity typical of other intracellular fungal pathogens. Differences in growth rates of intracellular Blastomyces yeast and the differences in amounts of yeast phagocytized by macrophages were determined for macrophages derived from peripheral blood monocytes from 11 persons with treated blastomycosis and 11 normal, healthy persons. Cellular immunity was examined by lymphocyte uptake of [3H]thymidine in response to a specific antigen of Blastomyces yeast. Yeast were more readily phagocytized by macrophages from the previously treated donors when compared with those from the normal donors; the yeast were confirmed to be intracellular by transmission electron microscopy. Likewise, a decrease in growth rates of yeast was demonstrable in cultures of macrophages from previously treated donors as compared with normal donors. This greater efficiency of phagocytosis and growth inhibition of B. dermatitidis reflects another mechanism of human cellular immunity to this fungal infection.
Blastomyces↗
Passive transfer of delayed hypersensitivity to Blastomyces dermatitidis between mice.
This study further characterized the delayed hypersensitivity state induced in animals by Blastomyces dermatitidis exposure. Passive transfer of delayed hypersensitivity by transfer of cells and inhibition of migration of peritoneal exudate cells were studied, using sensitized mice of two inbred strains. Donor mice were subcutaneously inoculated with viable B. dermatitidis yeast cells. After 15 days, spleen cells or serum from these animals were injected intravenously into normal recipients of the same strain. After 24 h these mice were footpad tested with killed B. dermatitidis yeast cell antigen. Mice receiving spleen cells from sensitized animals had a significant increase in footpad thickness 24 to 48 h after testing. Those receiving only serum remained negative. Migration of peritoneal exudate cells from blastomyces-sensitive donor mice was inhibited by presence of blastomycin but not by mycobacterial antigen. Neither blastomyces-sensitive nor control animals reacted to footpad or migration inhibition testing with mycobacterial antigen.
Animals↗
Growth inhibition of Blastomyces dermatitidis in alveolar and peripheral macrophages from patients with blastomycosis.
Blastomyces dermatitidis evokes a pyogranulomatous disorder with organisms frequently found inside giant cells. Macrophages from bronchoalveolar lavage fluid and peripheral blood in monolayer cell cultures were challenged with live yeast organisms to examine phagocytosis and intracellular growth. A greater number of macrophages from patients recovering from blastomycosis had phagocytized Blastomyces compared with macrophages from healthy control donors. No differences were detected within the groups between alveolar and peripheral macrophages. Intracellular growth of the fungus was reduced in cultures of both cell types from patients compared with those from control subjects. Supernatants from specific Blastomyces antigen-stimulated lymphocyte cultures were collected, and treatment with the supernatant to control donors' macrophages resulted in increased phagocytosis and inhibition of intracellular growth. Antigen-induced lymphocyte stimulation as a correlate of cellular immunity is qualitatively related to alveolar or peripheral macrophage phagocytosis and growth inhibition of this fungus.
Blastomyces↗
Identification of Blastomyces dermatitidis in the stool of a dog with acute pulmonary blastomycosis.
We report the identification of Blastomyces dermatitidis by microscopic examination of a direct faecal smear from a dog with pulmonary blastomycosis. A simultaneously obtained faecal culture grew Blastomyces dermatitidis. The fungus was also cultured from a transtracheal sample from this same dog. This report suggests that yeast-phase cells of B. dermatitidis may be recovered in the stool of dogs with pulmonary blastomycosis following transit through the gastrointestinal tract of swallowed infected sputum. Implications regarding the ecology of Blastomyces dermatitidis are discussed.
Animals↗
Non-specific incorporation of nucleic acid precursors in Blastomyces dermatitidis and Histoplasma capsulatum.
Incorporation of thymidine, thymidine monophosphate (TMP), thymidine triphosphate (TTP), uridine and orotic acid into DNA, RNA and protein in Blastomyces dermatitidis and Histoplasma capsulatum was studied utilizing a specific acid hydrolysis technique developed for these fungi. Thymidine was incorporated to the greatest extent (approximately 0.5% of added label) followed by uridine, orotic acid, TMP and TTP. In Blastomyces, uridine and orotic acid labeled primarily RNA. TMP and TTP labeled RNA, DNA and protein at nearly the same level. In Histoplasma RNA was labeled poorly by any of these precursors. TMP and TTP labeled DNA predominately and protein to a slightly lower level. Deoxyadenosine or uridine media supplements of 250 micrograms/ml did not enhance incorporation. All precursors tested were found to be nonspecific in that RNA, DNA and protein were labeled. All data indicate that neither RNA nor DNA synthesis can be specifically measured in whole cells or acid precipitates by any of these precursors. Specific radiometric monitoring with these isotopes therefore requires the separation of these macromolecules.
Animals↗
Antibodies to antigens of Histoplasma, Blastomyces and Candida in HIV patients and carriers in Nigeria.
Serum samples from 60 subjects with confirmed HIV-1 infection including 28 AIDS patients and 32 carriers were examined by immunodiffusion for precipitating antibodies to antigens of Histoplasma, Blastomyces and Candida. Seven of the subjects, four patients and three carriers, showed antibodies to histoplasmin prepared from mycelial cultural filtrate of Histoplasma capsulatum var. capsulatum and H. capsulatum var. duboisii although without any clinical signs of classical or African histoplasmosis. Another eight subjects comprising five patients and three carriers demonstrated antibodies to yeast cell antigen of Candida albicans; three of the patients had oral lesions clinically suggestive of Candida infection. None of the serum samples revealed antibodies to "A" antigen (yeast cell antigen) of Blastomyces dermatitidis.
AIDS-Related Opportunistic Infections↗
Effect of immunosuppressants on dogs exposed two and one-half years previously to Blastomyces dermatitidis.
Observation of 33 dogs exposed to Blastomyces dermatitidis over a 2 1/2 year period is reported. During the first 1 1/2 years six dogs died and 23 (77%) developed skin lesions. Clinical evidence of disease in all the animals then subsided and remained so through another year of observation. Nine animals were sacrificed after 120 weeks and several tissues were cultured from each one. The remaining 18 were given immunosuppressants--azathioprine, prednisone, and cyclophosphamide--for eight weeks prior to autopsy. About one-half of the dogs were found to be infected with Histoplasma capsulatum and all were negative for B. dermatitidis by culture. The dogs were thought to have been inadvertently exposed to H. capsulatum during the first eight weeks of observation from a nearby soil plot before being moved to a more remote shelter. Skin testing with histoplasmin and blastomycin and complement-fixation (CF) tests, performed at various intervals after exposure to B. dermatitidis, appeared to reflect the course of disease with the number of reactors to the H. capsulatum antigens remaining relatively high while the number of reactors to B. dermatitidis antigens fell with the passage of time. The blastomycin skin test results in these dogs, however, persisted far longer than positive Blastomyces CF results. These data suggest that most dogs exposed to B. dermatitidis exhibit clinical symptoms of disease after about 12 weeks, persisting for about one year. The majority of animals recover spontaneously and completely. With H. capsulatum, however, the infection can be subclinical for at least a two-year period.
Animals↗
Comparative studies on the detection of delayed dermal hypersensitivity in experimental animals with lysate and filtrate antigens of Blastomyces dermatitidis.
Yeast cell and mycelial lysate and filtrate antigens prepared in our laboratory from two strains of Blastomyces dermatitidis (canine isolate T-58 and human isolate Le) were evaluated with respect to the detection of delayed dermal hypersensitivity in hairless guinea pigs previously immunized with killed whole yeast cells from two strains of Blastomyces dermatitidis (T-58 and Le) and Histoplasma capsulatum (strain G-217A). The optimal potential reactivity (reactivity minus cross-reactivity) with regard to eliciting a dermal response in animals sensitized with B. dermatitidis was achieved with the yeast phase lysate and filtrate antigens prepared from both T-58 and Le isolates (mean axes of induration values ranging from 14.0 to 15.9 mm). In contrast, the mycelial phase reagents exhibited lower potential reactivity (mean axes of induration values ranging from 5.3 to 10.5 mm).
Animals↗
Cell wall studies of Histoplasma capsulatum and Blastomyces dermatitidis using autologous and heterologous enzymes.
Enzymes capable of hydrolyzing cell walls of Blastomyces dermatitidis and chemotypes I and II of Histoplasma capsulatum were prepared in the laboratory or obtained from commercial sources. They included chitinases, beta-1,3-glucanases, beta-1,6-glucanase, and Pronase. Monosaccharides and disaccharides of glucose released from the cell walls by the enzymes were determined qualitatively by paper and gas-liquid chromatography, and monosaccharides were quantitated by the latter technique as well. An enzyme system isolated from Streptomyces sp. containing both chitinase and glucanase released maximum amounts of glucose and N-acetylglucosamine from the cell walls of H. capsulatum chemotype I. A chitinase preparation, free of glucanase, from Serratia marcescens released only chitobiose and N-acetylglucosamine from chemotype I cell walls, but the total quantity of N-acetylglucosamine released was about 60% less than that released by the Streptomyces system. A beta-1,3-glucanase from Bacillus circulans hydrolyzed the cell walls of H. capsulatum chemotype I, but a beta-1,6-glucanase failed to release glucose from the same walls. Autolytic enzymes, viz., beta-1,3-glucanases and several glycosidases were detected as constitutive enzymes in both yeast and mycelial phases of B. dermatitidis and H. capsulatum chemotypes I and II. No difference in the amount of activity was found between cell sap and culture filtrate preparations. The beta-glucanases prepared from the Histoplasma and Blastomyces strains were active on the cell walls of the yeast phases of H. capsulatum chemotypes I and II, releasing laminaribiose and glucose, but were essentially inactive on the cell walls of B. dermatitidis. Chitinase, beta-1,6-glucanase, alpha-glucanase, and alpha-glucosidase activities were absent from these fungal enzyme preparations.
Acetylglucosamine↗
Use of an alkali-soluble water-soluble extract of Blastomyces dermatitidis yeast-phase cell walls and isoelectrically focused components in peripheral lymphocyte transformations.
An alkali-soluble water-soluble extract of Blastomyces dermatitidis yeast-phase cell walls was tested for its ability to elicit a response in lymphocytes isolated from the peripheral blood of Blastomyces-infected guinea pigs. Sequential preparations of the antigen were reproducible and specific in the in vitro lymphocyte transformation assay. Cross-reactivity of the antigen was not evident in lymphocyte transformation assays on lymphocytes obtained from Histoplasma-infected guinea pigs or from animals sensitized with complete Freund adjuvant. Fractionation of the antigen was accomplished on an isoelectric-focusing column, using a sucrose density gradient support. Components were assayed for activity in skin testing and lymphocyte transformation. Comparison of column fractions to the whole antigen showed greater response to the whole antigen in in vivo and in vitro assays.
Animals↗
WI-1, a novel 120-kilodalton surface protein on Blastomyces dermatitidis yeast cells, is a target antigen of cell-mediated immunity in human blastomycosis.
A large body of experimental data has demonstrated the central role of T cells in acquired resistance to the dimorphic fungus Blastomyces dermatitidis. We examined the human T-cell response to WI-1, a 120-kDa B. dermatitidis yeast cell surface protein recently shown to be an immunodominant antigen of the B-cell response in infected humans. Peripheral blood lymphocytes from 10 blastomycosis patients studied proliferated in response to WI-1 (mean, 19,431 cpm) and to the standard, crude cell wall antigen, Blastomyces alkali- and water-soluble antigen (B-ASWS) (mean, 19,131 cpm); lymphocytes from 10 histoplasmosis patients and 10 normal control subjects did not respond to WI-1. WI-1 stimulation of patient lymphocytes and rechallenge with WI-1 or B-ASWS showed that the antigens share immunodominant epitopes. Of 100 WI-1-responsive T-cell clones derived from peripheral blood, 10 were studied in detail to assess the phenotype, function, and ligands recognized. The clones exhibit the CD3+ CD4+ phenotype of helper T cells; 2 of 10 clones (and 21% of antigen-stimulated peripheral blood lymphocytes) use the V beta 8 T-cell receptor gene element to respond to WI-1. All the clones proliferate in response to both WI-1 and B-ASWS but not other fungal antigens, and some mediate potent cytolytic effects on WI-1- and B-ASWS-labeled targets. WI-1 recognition requires antigen processing and presentation of epitopes in association with HLA-DR (to noncytolytic clones) and HLA-DP (to cytolytic clones). From these findings, we conclude that CD4+ T cells with regulatory and cytolytic properties are involved in the development of acquired resistance of B. dermatitidis, that the cells are directed against WI-1, and that the manner of display of WI-1 peptide epitopes in conjunction with major histocompatibility complex class II may influence the profile of the immune response.
Antigens, Fungal↗