Identification of a Coccidioides immitis antigen 2 domain that expresses B-cell-reactive epitopes.
Antigen 2 (Ag2), a major immunoreactive component of Coccidioides immitis mycelium- and spherule-phase cell walls, was recently cloned in our laboratory and was shown to elicit T-cell responses in Coccidioides-immune mice. In this investigation, we evaluated recombinant Ag2 (rAg2) and PCR-generated Ag2 truncations for expression of B-cell-reactive epitopes in enzyme-linked immunosorbent and immunoblot assays with sera from patients with active coccidioidomycosis, a hyperimmune goat anti-Ag2 serum, and a murine anti-Ag2 monoclonal antibody that recognizes a conformational epitope. The results established that rAg2 expresses both linear and conformational B-cell-reactive epitopes which are localized to a domain comprised of amino acids 19 through 96 (designated A19-96). Truncations designed to identify epitopes within the A19-96 domain yielded fragments that either were nonreactive (A62-194, A19-61, and A49-79) or showed reduced reactivity (A19-79). Hence, A19-96 was the shortest domain expressing epitopes recognized by the panel of antibodies. The prevalence of antibodies to the A19-96 domain was evaluated in enzyme-linked immunosorbent assays of sera from 28 coccidioidomycosis patients. Antibody reactivity was detected in 79% of the patients' sera, and the level of antibody reactivity was directly correlated with disease severity. Whereas patients with pulmonary disease showed a mean response (A405) of 0.16 +/- 0.04, patients with disseminated coccidioidomycosis showed a mean response of 0.69 +/- 0.17 (P < 0.05). No reactivity was detected with sera from histoplasmosis or blastomycosis patients. The production of a recombinant peptide that expresses C. immitis-specific Ag2 epitopes provides a useful reagent for examining the role of anti-Ag2 antibodies in coccidioidomycosis.
Zhu
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Y
Department of Clinical Investigation, Texas Center for Infectious Disease, San Antonio 78223, USA.
Tryon
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Magee
D M
DM
Cox
R A
RA
D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood; D000946:Antigens, Fungal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D003045:Coccidioides / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D007114:Immunization; D051379:Mice
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D000939:Epitopes