Comparison of Coccidioides immitis arthrospore, mycelium, and spherule cell walls, and influence of growth medium on mycelial cell wall composition

Wheat, Robert W.; Tritschler, Catherine; Conant, Norman F.; Lowe, Edwin P.

doi:10.1128/iai.17.1.91-97.1977

Cited by 30 publications

(17 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Heat denaturation, usually associated with a loss of biological activity, generally breaks the ionic and covalent bonds that give proteins their secondary and tertiary struc- (32). The possible presence of cross-linked form of chitin-peptide in C. immitis cell walls was suggested by Wheat et al (34) from their finding that some glucosamine was lost during digestion with pronase. Electrophoresis and silver staining demonstrated 20 bands in the 28-h SE filtrate and 17 bands in the 120-h SE filtrate; mycelial F171 and SF had 5 and 4 bands, respectively.…”

Section: Discussionmentioning

confidence: 98%

Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis

Zimmer¹,

Pappagianis

1986

Infect Immun

View full text Add to dashboard Cite

The extracellular proteins produced by Coccidioides immitis during growth of the spherule-endospore-phase and mycelial-phase antigen (coccidioidin) were studied by polyacrylamide gel electrophoresis followed by immunoblot analysis to detect specific serologic function. Filtrates obtained from 28-and 120-h growth of the spherule-endospore phase were compared with each other and with coccidioidin by using negative, immunoglobulin M (IgM) precipitin-positive, or complement fixation-positive pooled and single human sera followed by peroxidase-labeled anti-human IgA, IgE, IgG, or IgM (heavy chain specific) or peroxidase-labeled concanavalin A to detect the reaction. A total of 35 bands was seen in the stained gels. Different patterns were noted among the two spherule-endospore preparations and unheated and heated coccidioidin. At least 15 electrophoretically separate antigens were detected with positive serum ranging in approximate molecular weight (Mr) from 100,000 to 18,000. Most were clustered between 45 and 60 kilodaltons (kDa). Common bands were noted at 48 and 18 kDa. At least one band at 48 kDa was strongly reactive with complement fixation-positive serum demonstrated by reaction with anti-IgG and anti-IgE. In contrast, doublet bands in the 50to 65-kDa area were highly reactive with IgM precipitin-positive serum detected by anti-IgM. IgM antibodies present in both positive sera reacted with a band at 46 kDa which was not reactive with IgG. Heating the antigens altered the reactivity of many of the antigens, including the 48-kDa band, but not the 46-kDa band. * Corresponding author. t Present address: American Micro/Scan, West Sacramento, CA 95691.

show abstract

Section: Discussionmentioning

confidence: 98%

Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis

Zimmer¹,

Pappagianis

1986

Infect Immun

View full text Add to dashboard Cite

show abstract

“…2C). Given the high content of chitin (5 to 22%) in mycelial Coccidioides immitis (29,30), the growth inhibition may have been due to blockage of septum formation as a result of reduced chitin synthesis. However, under the conditions of these assays, nikkomycin Z by itself is not fungicidal for most mycelial isolates of Coccidioides immitis at the concentrations tested and it does not readily affect the MLCs of azoles for most isolates.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Antifungal Activity of Nikkomycin Z in Combination with Fluconazole or Itraconazole

Rinaldi

1999

Antimicrob Agents Chemother

124

View full text Add to dashboard Cite

Nikkomycins are nucleoside-peptide antibiotics produced byStreptomyces species with antifungal activities through the inhibition of chitin synthesis. We investigated the antifungal activities of nikkomycin Z alone and in combination with fluconazole and itraconazole. Checkerboard synergy studies were carried out by a macrobroth dilution procedure with RPMI 1640 medium at pH 6.0. At least 10 strains of the following fungi were tested: Candida albicans, other Candida spp., Cryptococcus neoformans, Coccidioides immitis,Aspergillus spp., and dematiacious fungi (includingExophiala jeanselmei, Exophiala spinifera,Bipolaris spicifera, Wangiella dermatitidis,Ochroconis humicola, Phaeoannellomyces werneckii, and Cladophialophora bantiana), and 2 strains each of Fusarium, Scedosporium,Paecilomyces, Penicillium, andTrichoderma spp. A total of 110 isolates were examined. Inocula of fungal elements were standardized by hemacytometer counting or spectrophotometrically. MICs and minimum lethal concentrations (MLCs) were determined visually by comparison of growth in drug-treated tubes with growth in drug-free control tubes. Additive and synergistic interactions between nikkomycin and either fluconazole or itraconazole were observed against C. albicans, Candida parapsilosis, Cryptococcus neoformans, andCoccidioides immitis. Marked synergism was also observed between nikkomycin and itraconazole against Aspergillus fumigatus and Aspergillus flavus. No antagonistic interaction between the drugs was observed with any of the strains tested.

show abstract

“…Previous investigations have demonstrated that methylated mannose-containing polysaccharides occur in the walls of arthroconidia, spherules, and hyphae of C. immitis (3,31,(35)(36)(37). In addition, previous reports have indicated that the ID-TP antigen is predominantly polysaccharide (13, 17, 41), and partial to total loss of reactivity with IgM precipitin occurs when the antigen is subjected to periodate oxidation (13).…”

Section: 'mentioning

confidence: 99%

“…We also show that reactivity of the antigen with serum samples from patients with coccidioidomycosis is retained after pronase digestion or heating (100°C, 5 min) but is lost after periodate oxidation. Examination of the carbohydrate composition of the TP antibody-reactive antigen (TP-Ag) by gas chromatography-mass spectroscopy revealed 3-0-methylmannose (3-0-MM), a sugar that was previously shown to be a component of immunoreactive fractions of C. immitis (3,15,17,28,(35)(36)(37) and possibly unique to this pathogen among the systemic fungal pathogens (3,37). In this report, we also further examine the serologic reactivity of the 120-kDa TP-Ag and, in particular, the significance of 3-0-MM in precipitin binding to this macromolecule.…”

mentioning

confidence: 99%

Composition, serologic reactivity, and immunolocalization of a 120-kilodalton tube precipitin antigen of Coccidioides immitis

et al. 1990

Self Cite

View full text Add to dashboard Cite

Diagnosis of coccidioidomycosis largely depends on serologic tests. In this investigation, the enzyme-linked immunosorbent assay (ELISA) was used to detect patient immunoglobulin M (IgM) precipitin antibody binding to a 120-kilodalton (kDa) fraction previously isolated from an alkali-soluble, water-soluble extract of the arthroconidial wall and mycelial culture filtrate plus toluene lysate of Coccidioides immitis. Results of the serologic response to this tube precipitin antigen (TP-Ag) in the ELISA correlated well with results of immunodiffusion assays of 30 serum samples from patients. Immunoelectron microscopic examinations of arthroconidia and spherules were performed with patient IgM precipitin antibodies isolated from sera eluted over a solid-phase immunosorbent column containing the purified 120-kDa TP-Ag. The antibody probe located the 120-kDa TP-Ag on the walls of in vitro-grown arthroconidia and spherules. Pronase digestion and heating (100°C, 5 min) had no apparent effect on the activity of the 120-kDa TP-Ag, while periodate oxidation resulted in total loss of its immunodiffusion-TP activity. Analysis of the carbohydrate composition of the TP-Ag revealed xylose, 3-0-methylmannose (3-0-MM), mannose, galactose, and glucose. Competitive inhibition ELISAs were used to demonstrate that 3-0-MM is largely responsible for the reactivity of IgM precipitin antibodies with the 120-kDa TP-Ag. Synthetic 3-0-MM may be a useful probe for detection of anti-Coccidioides precipitin antibodies in the ELISA.

show abstract

Comparison of Coccidioides immitis arthrospore, mycelium, and spherule cell walls, and influence of growth medium on mycelial cell wall composition

Cited by 30 publications

References 23 publications

Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis

Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis

In Vitro Antifungal Activity of Nikkomycin Z in Combination with Fluconazole or Itraconazole

Composition, serologic reactivity, and immunolocalization of a 120-kilodalton tube precipitin antigen of Coccidioides immitis

Contact Info

Product

Resources

About