Extraction of serologic and delayed hypersensitivity antigens from spherules of Coccidioides immitis

Galgiani, John N.; Dugger, Kris O.; Ampel, Neil M.; Sun, Sung H.; Law, John H.

doi:10.1016/0732-8893(88)90075-2

Cited by 23 publications

(22 citation statements)

References 37 publications

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“…We have shown that toluene-induced autolysis at 350C of mature spherules in vitro also yields antigens active in the IDTP and IDCF reactions. By exposing first-generation spherules grown in vitro to toluene at 40TC, Galgiani et al (39) produced an antigen reactive by IDTP but lacking IDCF activity. Antigens useful in serologic tests for coccidioidomycosis are available from a number of commercial sources.…”

Section: Antibodymentioning

confidence: 99%

Serology of coccidioidomycosis

1990

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Serologic tests have assisted in the diagnosis and prognosis of coccidioidomycosis for a half-century. The causative agent, Coccidioides immitis, is a dimorphic fungus existing in a hyphal form with arthroconidia in nature and in the usual culture. The arthroconidia represent the inhaled infective forms which in vivo and under special laboratory conditions form spherules which endosporulate. The culture filtrate/autolysate (coccidioidin) from the hyphal phase has provided antigens of suitable reliability for currently used serologic tests. These tests are primarily to determine the two major antibody responses: the early immunoglobulin M (IgM) response is useful in the diagnosis of acute primary coccidioidomycosis. Later, IgG is produced and usually outlasts the IgM, persisting in chronic coccidioidomycosis. The IgM is detectable by tube precipitin, a corresponding immunodiffusion, or latex particle agglutination tests. The pertinent antigen(s) is heat stable and pronase resistant and appears to be largely carbohydrate, mainly mannose with some 3-O-methyl mannose. The IgG detectable in the serum and other body fluids by complement fixation and a corresponding immuno-diffusion is useful in diagnosis, and its quantitation provides an indicator of progression of disease (increasing titer) or regression (decreasing titer). The pertinent antigen appears to be a heat-labile, pronase-sensitive protein which in an unreduced form has a molecular weight of 110,000. A third very useful serologic procedure is the exoantigen test for identification of putative cultures of C. immitis.

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Section: Antibodymentioning

confidence: 99%

Serology of coccidioidomycosis

1990

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“…rAg2/PRA (1) was used with either an oil emulsion of monophosphoryl lipid A (MPL-SE) for subcutaneous injection or aqueous MPL (MPL-AF) for intranasal administration (Corixa, Inc., Hamilton, Mont.). Whole-spherule vaccine was prepared from 96-h growths of arthroconidia in liquid Converse medium, resulting in mature endosporulated spherules (11,12). After exposure to 5% formalin, washed spherules were lyophilized.…”

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confidence: 99%

Protection of Mice againstCoccidioides immitisIntranasal Infection by Vaccination with Recombinant Antigen 2/PRA

et al. 2002

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Subcutaneous vaccination with recombinant antigen 2/PRA (rAg2/PRA) protected BALB/c mice against intranasal infection with Coccidioides immitis. Subcutaneously vaccinated C57BL/6 mice and intranasally vaccinated BALB/c mice were protected against larger numbers of infecting spores. Weight loss correlated with lethality, but histologic appearance did not. These studies support rAg2/PRA vaccination to prevent coccidioidomycosis.

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“…For whole spherules, it seems most plausible that surface components react in some way with hypochlorous acid to render it ineffective. For the spherule extracts, gas chromatographic analysis has shown them to contain glucose, mannose, N-acetyl-glucosamine, O-methylmannose and galactosamine [ 17]. All of these sugars are prominent constituents of the spherule wall [ 17].…”

Section: Changes In Oxidant Killing Related To Larger Inoculum Sizementioning

confidence: 99%

“…To prepare coccidioidal substances for these studies, mature spherules were extracted by exposure to 3% toluene as described previously [17]. Further fractionation was accomplished by ion exchange chromatography using DEAE Biogel (Bio-Rad, Richmond, CA) at pH 7.0 and eluted with a sodium chloride gradient of 0-0-1-0 i.…”

Section: Microbial Killing Assaymentioning

confidence: 99%

Killing ofCoccidioides immitisby hypochlorous acid or monochloramine

Galgiani¹

1990

Med Mycol

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To identify possible explanations for the resistance of Coccidioides immitis to killing by human neutrophils, its susceptibility to typical oxidants generated during the neutrophil respiratory burst was compared to the sensitivity of other microbes. When microbial suspensions were exposed to hypochlorous acid, arthroconidia or spherules of C. immitis were killed more slowly than yeast cells of Candida (Torulopsis) glabmta or Staphylococcus aureus. In contrast, exposure to the more lipophilic oxidant, monochloramine, produced equivalent rates of killing for spherules and yeast cells of C. glabrata, and arthroconidia were killed more rapidly. When larger microbial concentrations were used, significantly greater concentrations of hypochlorous acid were required to kill equivalent percentages of spherules as compared to yeast cells of C. glabrata. Mixing studies with either whole spherules or extracts of spherules demonstrated that these substances could also block hypochlorous acid killing of S. aureus. These studies suggest possible mechanisms whereby C. immitis might resist oxidative phagocytic killing. Coccidioidomycosis, when contracted by otherwise healthy persons, is typically a self-limited infection [13, 23]. The critical host defense responsible for resolution of the illness is believed to be T lymphocyte-dependent cytokine-mediated activation of tissue macrophages analogous to control mechanisms identified in experimental studies of murine infection [2-5]. However, various clinical observations stronglysuggest that control may not require lethal eradication of the fungus in humans. These include the high frequency of recovery of Coccidioides imrnitis from surgically resected quiescent pulmonary nodules [30], chronic asymptomatic excretion of C. immitis in certain patients from pulmonary cavities or prostatic foci [9, 40], and the recrudescence of infection in immunocompromized patients [6, 10, 38] many years after the initial exposure and often far from the endemic area. Furthermore, we and others have noted the striking resistance of C. immitis to the normally microbicidal effect of human neutrophils [ 15, 16, 19]. The reasons for this failure of neutrophil killing have not been readily apparent since other observations have indicated that neutrophils are

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Extraction of serologic and delayed hypersensitivity antigens from spherules of Coccidioides immitis

Cited by 23 publications

References 37 publications

Serology of coccidioidomycosis

Serology of coccidioidomycosis

Protection of Mice againstCoccidioides immitisIntranasal Infection by Vaccination with Recombinant Antigen 2/PRA

Killing ofCoccidioides immitisby hypochlorous acid or monochloramine

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