Immunoaffinity isolation and partial characterization of the Coccidioides immitis antigen detected by the tube precipitin and immunodiffusion-tube precipitin tests

Zimmer, Barbara; Pappagianis, Demosthenes

doi:10.1128/jcm.27.8.1759-1766.1989

Cited by 22 publications

(11 citation statements)

References 35 publications

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“…Earlier reports of C. immitis antigens capable of eliciting a TP antibody response in patients identified the reactive macromolecules by a wide range of molecular sizes (21 to 330 kDa) (2,11,12,37). The ConA-bound material of the F+L fraction was previously shown to contain both the 120and 110-kDa TP-Ags (17).…”

Section: Discussionmentioning

confidence: 99%

“…Numerous preparations of C. immitis antigens have been used for assays which detect TP antibody in patients including the mycelial culture filtrate, filtrate plus toluene lysate of the isolated mycelia (F+L fraction), autolysate of the mycelial phase and parasitic cells (spherules plus endospores), and alkali-soluble, water-soluble extracts of isolated mycelial, conidial, and spherule wall fractions (10,12,17,24,37). In a recent study it was shown that concanavalin A (ConA) separation of the spherule culture filtrate permitted isolation of an affinity-bound fraction rich in TP antigen and with relatively few contaminating antigens (12a).…”

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confidence: 99%

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Antigen complex of Coccidioides immitis which elicits a precipitin antibody response in patients

1991

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The occurrence in patients of elevated levels of immunoglobulin M (IgM) precipitin antibody to Coccidioides immitis antigens, which are commonly detected by the immunodiffusion-tube precipitin (TP) assay, is suggestive of primary nondisseminating coccidioidomycosis. We previously demonstrated that the concanavalin Abound mycelial culture filtrate plus lysate preparation is a source of at least two TP antibody-reactive antigens (TP-Ags), which were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as 120and 110-kDa fractions. Evidence is presented here that the crude filtrate plus lysate preparation contains additional lectin-bound, TP antibody-reactive fractions as well as a component which elicits a complement fixation antibody response in patients. The 120and 110-kDa fractions were isolated from the antigen complex and further characterized in this paper. Both TP-Ags are glycoproteins and have been shown by immunoelectron microscopy to be colocalized within cytoplasmic vesicles and the wall of spherules. Deglycosylation of these TP-Ags by sodium periodate treatment resulted in a loss in patients of 82 to 95% of IgM adsorption to the antigens as detected by the enzyme-linked immunosorbent assay (ELISA). Comparison of their carbohydrate compositions revealed that mannose and glucose are the predominant monosaccharides of both TP-Ags but only the 120-kDa fraction contained 3-O-methylmannose, a sugar which appears to be unique to C. immitis among the systemic fungal pathogens. We previously showed that 3-O-methylmannose is at least partly responsible for the reactivity of IgM antibody with the 120-kDa TP-Ag. Good correlation was shown between results of immunodiffusion-TP assays and ELISAs of IgM response to both the 120and 110-kDa fractions by using 70 serum samples from patients with proved coccidioidomycosis. However, only 2.8% (3 of 109) of the serum samples from patients with other mycoses and nonmycotic infections showed IgM adsorption to the 120-kDa TP-Ag as detected by the ELISA, while 21.1% (23 of 109) showed IgM adsorption to the 110-kDa TP-Ag. The 120-kDa TP-Ag is a potentially valuable serodiagnostic reagent for detection of specific IgM by ELISA in patients with primary coccidioidomycosis.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Antigen complex of Coccidioides immitis which elicits a precipitin antibody response in patients

1991

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“…The TP antibody has been shown to be of the immunoglobulin M (IgM) class (30). Several investigators have described precipitin-reactive fractions of C. immitis which were characterized as glycoproteins with molecular sizes ranging from 50 to 330 kilodaltons (kDa) (13,16,17,39,41). We have reported the isolation of an arthroconidialand mycelial culture filtrate plus lysate-derived 120-kDa glycoprotein of C. immitis by gel filtration chromatography which was reactive with antibody from patients with coccidioidomycosis in both the TP test and ID-TP assay (24).…”

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confidence: 99%

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

et al. 1990

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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.

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“…Both Ags contain 3-0-MM residues, which in the case of the 120-kDa TP-Ag have been shown to be at least partly responsible for reactivity of specific IgM antibody with the glycoprotein (6,7). An immunoaffinity-chromatography-isolated and partially characterized TP-Ag, with molecular sizes of 140 and 225 kDa under reducing and nonreducing conditions, respectively, was also reported to contain 3-0-MM (43). Periodate oxidation of the 3-glucosidase yielded a 47-kDa polypeptide detected by SDS-PAGE under reducing conditions.…”

Section: Discussionmentioning

confidence: 98%

A seroreactive 120-kilodalton beta-1,3-glucanase of Coccidioides immitis which may participate in spherule morphogenesis

Kruse

Cole

1992

Infect Immun

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A 13-glucosidase of Coccidioides immitis was identified in electrophoresis gel separations of the concanavalin Abound mycelial culture-filtrate-plus-lysate preparation. p-Nitrophenol-1-D-glucopyranoside was used as the substrate to visualize the enzymatically active fraction in nonreducing gels. The gel-isolated, chromatographically purified enzyme has an optimal pH of 8.0 and cleaves 0-1,3-glycosyl linkages. The alkaline 13-glucosidase was further characterized by a pI of 3.8 to 4.0, optimal activity at 37 to 40°C, and molecular size of 120 kDa as identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified 1-glucosidase is identical to a previously reported 120-kDa antigen (Ag) which reacts with immunoglobulin M (IgM) tube precipitin (TP) antibody in sera from patients with coccidioidomycosis. The TP-Ag was described as a valuable serodiagnostic reagent for detection of specific IgM in patients with early coccidioidal infections. The * Corresponding author. enzyme reacts with immunoglobulin M (IgM) precipitin antibody in sera from patients with coccidioidomycosis.

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Immunoaffinity isolation and partial characterization of the Coccidioides immitis antigen detected by the tube precipitin and immunodiffusion-tube precipitin tests

Cited by 22 publications

References 35 publications

Antigen complex of Coccidioides immitis which elicits a precipitin antibody response in patients

Antigen complex of Coccidioides immitis which elicits a precipitin antibody response in patients

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

A seroreactive 120-kilodalton beta-1,3-glucanase of Coccidioides immitis which may participate in spherule morphogenesis

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