Abstract
We have previously cloned the cDNA fragment that encodes the complement fixation antigen of Coccidioides immitis. The recombinant protein was highly sensitive in detecting CF antibody in sera from patients with coccidioidomycosis but was not specific to C. immitis, as evidenced by its reactivity with sera from patients with histoplasmosis and, to lesser extent, blastomycosis. We undertook this study to determine if the epitope(s) that reacts with CF antibody is the same or differs from the epitopes that are shared with Histoplasma capsulatum and Blastomyces dermatitidis. PCR-generated CF/chitinase cDNA fragments were cloned and examined for their reactivity in enzyme-linked immunosorbent assays using sera from patients with coccidioidomycosis, histoplasmosis, or blastomycosis. A peptide domain comprised of amino acid residues 20 through 310 was shown to express an epitope(s) that is specific to anti-Coccidioides CF antibody. The peptide detected serum antibody in 21 (95%) of 22 patients with active coccidioidomycosis and was without reactivity with sera from 20 patients with histoplasmosis, 15 patients with blastomycosis, and 14 healthy subjects. Antibody titers to the recombinant peptide directly correlated with CF antibody titers (P < 0.01), and preadsorption of reference CF antiserum with the peptide ablated the reactivity of the antiserum in the immunodiffusion assay for CF antibody. The delineation of a recombinant peptide that has both sensitivity and specificity will provide a valuable tool for detecting CF antibody and for evaluating the role of CF antibody in the host response to C. immitis.
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Selected References
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- Beaman L. V., Pappagianis D., Benjamini E. Mechanisms of resistance to infection with Coccidioides immitis in mice. Infect Immun. 1979 Mar;23(3):681–685. doi: 10.1128/iai.23.3.681-685.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowen A. R., Chen-Wu J. L., Momany M., Young R., Szaniszlo P. J., Robbins P. W. Classification of fungal chitin synthases. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):519–523. doi: 10.1073/pnas.89.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowman B. H., Taylor J. W., White T. J. Molecular evolution of the fungi: human pathogens. Mol Biol Evol. 1992 Sep;9(5):893–904. doi: 10.1093/oxfordjournals.molbev.a040766. [DOI] [PubMed] [Google Scholar]
- Cox R. A. Antigenic structure of Coccidioides immitis. Immunol Ser. 1989;47:133–170. [PubMed] [Google Scholar]
- Cox R. A., Britt L. A., Michael R. A. Isolation of Coccidioides immitis F antigen by immunoaffinity chromatography with monospecific antiserum. Infect Immun. 1987 Jan;55(1):227–232. doi: 10.1128/iai.55.1.227-232.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dolan M. J., Cox R. A. Production and characterization of a monoclonal antibody to the complement fixation antigen of Coccidioides immitis. Infect Immun. 1991 Jun;59(6):2175–2180. doi: 10.1128/iai.59.6.2175-2180.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Emini E. A., Hughes J. V., Perlow D. S., Boger J. Induction of hepatitis A virus-neutralizing antibody by a virus-specific synthetic peptide. J Virol. 1985 Sep;55(3):836–839. doi: 10.1128/jvi.55.3.836-839.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galgiani J. N. Coccidioidomycosis. West J Med. 1993 Aug;159(2):153–171. [PMC free article] [PubMed] [Google Scholar]
- Jameson B. A., Wolf H. The antigenic index: a novel algorithm for predicting antigenic determinants. Comput Appl Biosci. 1988 Mar;4(1):181–186. doi: 10.1093/bioinformatics/4.1.181. [DOI] [PubMed] [Google Scholar]
- Johnson S. M., Pappagianis D. The coccidioidal complement fixation and immunodiffusion-complement fixation antigen is a chitinase. Infect Immun. 1992 Jul;60(7):2588–2592. doi: 10.1128/iai.60.7.2588-2592.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson S. M., Zimmermann C. R., Pappagianis D. Amino-terminal sequence analysis of the Coccidioides immitis chitinase/immunodiffusion-complement fixation protein. Infect Immun. 1993 Jul;61(7):3090–3092. doi: 10.1128/iai.61.7.3090-3092.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson S. M., Zimmermann C. R., Pappagianis D. Use of a recombinant Coccidioides immitis complement fixation antigen-chitinase in conventional serological assays. J Clin Microbiol. 1996 Dec;34(12):3160–3164. doi: 10.1128/jcm.34.12.3160-3164.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klein B. S., Jones J. M. Isolation, purification, and radiolabeling of a novel 120-kD surface protein on Blastomyces dermatitidis yeasts to detect antibody in infected patients. J Clin Invest. 1990 Jan;85(1):152–161. doi: 10.1172/JCI114406. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kong Y. M., Savage D. C., Levine H. B. Enhancement of immune responses in mice by a booster injection of Coccidioides spherules. J Immunol. 1965 Dec;95(6):1048–1056. [PubMed] [Google Scholar]
- Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
- Pan S., Sigler L., Cole G. T. Evidence for a phylogenetic connection between Coccidioides immitis and Uncinocarpus reesii (Onygenaceae). Microbiology. 1994 Jun;140(Pt 6):1481–1494. doi: 10.1099/00221287-140-6-1481. [DOI] [PubMed] [Google Scholar]
- Pappagianis D., Lindsay S., Beall S., Williams P. Ethnic background and the clinical course of coccidioidomycosis. Am Rev Respir Dis. 1979 Oct;120(4):959–961. doi: 10.1164/arrd.1979.120.4.959. [DOI] [PubMed] [Google Scholar]
- Pappagianis D., Zimmer B. L. Serology of coccidioidomycosis. Clin Microbiol Rev. 1990 Jul;3(3):247–268. doi: 10.1128/cmr.3.3.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SMITH C. E., SAITO M. T., BEARD R. R., KEPP R. M., CLARK R. W., EDDIE B. U. Serological tests in the diagnosis and prognosis of coccidioidomycosis. Am J Hyg. 1950 Jul;52(1):1–21. doi: 10.1093/oxfordjournals.aje.a119404. [DOI] [PubMed] [Google Scholar]
- SMITH C. E., SAITO M. T., SIMONS S. A. Pattern of 39,500 serologic tests in coccidioidomycosis. J Am Med Assoc. 1956 Feb 18;160(7):546–552. doi: 10.1001/jama.1956.02960420026008. [DOI] [PubMed] [Google Scholar]
- Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
- Sternberg S. The emerging fungal threat. Science. 1994 Dec 9;266(5191):1632–1634. doi: 10.1126/science.7702654. [DOI] [PubMed] [Google Scholar]
- Van Regenmortel M. H. Structural and functional approaches to the study of protein antigenicity. Immunol Today. 1989 Aug;10(8):266–272. doi: 10.1016/0167-5699(89)90140-0. [DOI] [PubMed] [Google Scholar]
- Yang C., Zhu Y., Magee D. M., Cox R. A. Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen. Infect Immun. 1996 Jun;64(6):1992–1997. doi: 10.1128/iai.64.6.1992-1997.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yang M. C., Magee D. M., Kaufman L., Zhu Y., Cox R. A. Recombinant Coccidioides immitis complement-fixing antigen: detection of an epitope shared by C. immitis, Histoplasma capsulatum, and Blastomyces dermatitidis. Clin Diagn Lab Immunol. 1997 Jan;4(1):19–22. doi: 10.1128/cdli.4.1.19-22.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zimmermann C. R., Johnson S. M., Martens G. W., White A. G., Pappagianis D. Cloning and expression of the complement fixation antigen-chitinase of Coccidioides immitis. Infect Immun. 1996 Dec;64(12):4967–4975. doi: 10.1128/iai.64.12.4967-4975.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
