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Journal of Clinical Microbiology, November 2001, p. 3830-3837, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3830-3837.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Characterization of AFMP1: a Novel Target for Serodiagnosis of Aspergillosis

Kwok-Yung Yuen,1,2,* Che-Man Chan,1 King-Man Chan,1 Patrick C. Y. Woo,1 Xiao-Yan Che,1 Andy S. P. Leung,1 and Liang Cao1

Department of Microbiology, The University of Hong Kong, Queen Mary Hospital,1 and HKU-Pasteur Research Centre,2 Hong Kong

Received 31 January 2001/Returned for modification 21 July 2001/Accepted 29 July 2001

We cloned the AFMP1 gene, which encodes the first antigenic cell wall galactomannoprotein in Aspergillus fumigatus. AFMP1 codes for a protein, Afmp1p, of 284 amino acid residues, with a few sequence features that are present in Mp1p, the antigenic cell wall mannoprotein in Penicillium marneffei that we described previously, as well as several other cell wall proteins of Saccharomyces cerevisiae and Candida albicans. It contains a serine- and threonine-rich region for O glycosylation, a signal peptide, and a putative glycosylphosphatidyl inositol attachment signal sequence. Specific anti-Afmp1p antibody was generated with recombinant Afmp1p protein purified from Escherichia coli to allow further characterization of Afmp1p. Afmp1p has a high affinity for Galanthus nivalis agglutinin, a characteristic indicative of a mannoprotein. Furthermore, it was recognized by a rat monoclonal antibody against the galactofuran side chain of galactomannan, indicating that it is a galactomannoprotein. Ultrastructural analysis by immunogold staining indicated that Afmp1p is present in the cell walls of the hyphae and conidia of A. fumigatus. Finally, it was observed that patients with aspergilloma and invasive aspergillosis due to A. fumigatus develop a specific antibody response against Afmp1p. This suggested that the recombinant protein and its antibody may be useful for serodiagnosis in patients with aspergilloma or invasive aspergillosis, and the protein may represent a good cell surface target for host humoral immunity.

* Corresponding author. Mailing address: Department of Microbiology, The University of Hong Kong, University Pathology Building, Queen Mary Hospital, Hong Kong. Phone: (852) 28554892. Fax: (852) 28551241. E-mail: microgen{at}hkucc.hku.hk.

Journal of Clinical Microbiology, November 2001, p. 3830-3837, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3830-3837.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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