Antigenic heterogeneity of an alkali-soluble, water-soluble cell wall extract of Coccidioides immitis

Cox, Rebecca A.; Britt, L A

doi:10.1128/iai.50.2.365-369.1985

Cited by 25 publications

(20 citation statements)

References 16 publications

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“…Earlier IEP studies of SCWF have revealed that at least some of the antigenic components of this fraction correspond to Ags in the anti-CDN reference system (6). CDN has been shown to be effective in eliciting delayed-type hypersensitivity responses, detecting IgE antibody to C. immitis, and reacting with anti-Coccidioides TP and CF antibodies (13), and has been used for many years as an immunodiagnostic Ag (17). Immunodiffusion studies of SCWF have shown that the conidial fraction is reactive with both TP and CF antibodies, which are the primary immunodiagnostic tests for this systemic mycosis (17).…”

Section: Discussionmentioning

confidence: 99%

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An immunoreactive, water-soluble conidial wall fraction of Coccidioides immitis

Cole¹,

Kirkland²,

Sun³

1987

Infect Immun

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Arthroconidia stripped of their outer, hydrophobic wail layer release an immunoreactive, water-soluble fraction (SCWF), the composition of which is examined in this paper. The immunogenicity of SCWF was determined by its reactivity with anti-Coccidioides immitis complement-fixing antibody and tube precipitin antibody in a standardized immunodiffusion assay as well as reactivity in a lymphocyte proliferation assay. SCWF was shown to be more immunoreactive for immune lymphocytes than selected cytosol and culture supernatant fractions of C. immitis. Rabbit antisera raised against SCWF were used in immunoelectronmicroscopic and immunofluorescence studies to confirm that the immunoreactive components of SCWF are primarily associated with the inner conidial wall. The antigenic composition of the conidial wall fraction was characterized by advancing-line immunoelectrophoresis with the previously established coccidioidin reference system. Protein composition was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Chromatographic fractionation of SCWF was performed on a Sephacryl S-300 preparative column. Selected fractions which showed significantly higher immunoreactivity and less complex antigenic composition than whole SCWF were characterized. Two heat-sensitive antigens of Sephacryl fraction 3a were identified. One or both of these antigens may correspond to the complement-fixing antigen, which is potentially important as an immunodiagnostic antigen. Fraction 4 contained a previously described wall-associated antigen (AgCS) which may also be of immunodiagnostic value. We conclude that the conidial envelope is a reservoir of immunoreactive macromolecules which may play significant roles in early stages of infection.

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Section: Discussionmentioning

confidence: 99%

“…16) and FR5 (not shown) revealed the absence of Ag2, a common component of SCWF and FRs 2, 3a, and 3b. Cox and co-workers have proposed that a heterogeneous alkali-soluble, water-soluble wall extract of C. immitis also contains Ag2 (13,14). A common antigen of both FRs 4 and 5 is AgCS (Fig.…”

Section: Discussionmentioning

confidence: 99%

An immunoreactive, water-soluble conidial wall fraction of Coccidioides immitis

Cole¹,

Kirkland²,

Sun³

1987

Infect Immun

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“…The development of a sensitive and specific immunoassay, such as an enzyme-linked immunosorbent assay, has been thwarted by the lack of a highly purified antigen. In previous studies from this laboratory, we isolated the CF antigen from coccidioidin (CDN) by using a combination of physiochemical procedures and produced a polyclonal goat anti-CF serum and a murine IgG1 monoclonal antibody (MAb) directed against a CF-specific epitope (4,8). The anti-CF antibodies proved to be effective ligands for isolating the antigen directly from CDN, but because of their high affinity, problems were encountered in recovering the antigen in a biologically active form (3).…”

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confidence: 99%

Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen

et al. 1996

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Detection of anti-Coccidioides complement-fixing (CF) antibody is a valuable diagnostic and prognostic aid in coccidioidomycosis. The CF antibody response is directed against a heat-labile antigen that has chitinase activity, hereafter referred to as the CF/chitinase protein. To identify and clone this immunoreactive enzyme, we constructed a Coccidioides immitis cDNA lambda ZAP expression library from spherule RNA and detected fusion peptides expressing CF epitopes by immunoscreening. A cDNA clone consisting of 1,623 bp was identified, sequenced, and found to contain a single open reading frame that encodes a protein of 47 kDa with 427 amino acids. Deduced amino acid sequence analyses showed that the cloned CF/chitinase cDNA contains a 35-amino-acid region, beginning at Ser-18 and ending at Arg-52, which has 92% homology with the reported N-terminal amino acid sequence of authentic CF/chitinase protein. The first 17 amino acids in the deduced sequence of the cloned cDNA are not present on the mature CF/chitinase protein, suggesting that it may be a signal peptide. Expression of the CF/chitinase cDNA insert by using the pGEX-4T-3 vector yields a fusion peptide that bears CF-specific epitopes and shows chitinase activity. The CF/chitinase clone will enable large-scale production of the recombinant CF antigen for use in immunoassays and facilitate studies on the role of chitinase in the morphogenesis of C. immitis.

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“…2fD-IEP. The techniques of 2D-IEP were performed according to methods detailed in earlier reports (3,4). Hyperimmune goat anti-CDN was the same antiserum used previously (3,4) and, before use in this study, was shown to be reactive with the F antigen, as assessed in the IDCF assay.…”

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confidence: 99%