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Research Article Free access | 10.1172/JCI114406

Isolation, purification, and radiolabeling of a novel 120-kD surface protein on Blastomyces dermatitidis yeasts to detect antibody in infected patients.

B S Klein and J M Jones

Department of Pediatrics, University of Wisconsin Medical School, University of Wisconsin Hospital and Clinics 53792.

Find articles by Klein, B. in: PubMed | Google Scholar

Department of Pediatrics, University of Wisconsin Medical School, University of Wisconsin Hospital and Clinics 53792.

Find articles by Jones, J. in: PubMed | Google Scholar

Published January 1, 1990 - More info

Published in Volume 85, Issue 1 on January 1, 1990
J Clin Invest. 1990;85(1):152–161. https://doi.org/10.1172/JCI114406.
© 1990 The American Society for Clinical Investigation
Published January 1, 1990 - Version history
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Abstract

No well-defined Blastomyces-specific antigens are currently available. We used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting to identify immunologically active molecules in the cell wall of B. dermatitidis. A major immunoreactive 120-kD protein (WI-1) was present in all five strains studied and comprised 5% of the protein in the cell wall extract obtained after freezing and thawing yeast cells. WI-1 was recognized by serum from all 10 patients with blastomycosis but by none of those from 5 patients with histoplasmosis. It was purified by electroelution, radiolabeled with 125I, and incorporated into a radioimmunoassay (RIA) for serodiagnosis of blastomycosis. Antibody to WI-1 was detected in 58 (85%) of 68 patients with blastomycosis (geometric mean titer, 1:2,981), in two (3%) of 73 patients with histoplasmosis, coccidioidomycosis, sporotrichosis, or candidiasis (titers, 1:86 and 1:91) and in none of 44 healthy persons. WI-1 was shown to be a surface molecule abundant on B. dermatitidis yeasts that were indirectly stained with serum from a rabbit immunized with WI-1. Approximately 0.93 pg of WI-1 or 4.7 x 10(6) WI-1 molecules were found on the surface of an individual yeast using an antigen-inhibition RIA; none was found on Histoplasma capsulatum or Candida albicans yeasts. We conclude that WI-1 is a novel, immunologically active surface molecule on the invasive form of B. dermatitidis and that WI-1 can be used to reliably detect antibody and study the immunopathogenesis of blastomycosis.

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