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Use of recombinant mitogillin for improved serodiagnosis of Aspergillus fumigatus-associated diseases.

Abstract
During human infection, Aspergillus fumigatus secretes a 18-kDa protein that can be detected as an immunodominant antigen in the urine of infected patients. Recently, this protein was shown to be mitogillin, a ribotoxin that cleaves a single phosphodiester bond of the 29S rRNA of eukaryotic ribosomes. We proved the immunogenic capacity of mitogillin in a rabbit animal model, indicating its usefulness as an antigen for serological diagnosis of invasive aspergillosis. The mitogillin gene from A. fumigatus was transferred from plasmid pMIT+ to expression vector pQE30 and expressed in Escherichia coli as a fusion protein. Purified recombinant mitogillin was recognized by serum immunoglobulin G (IgG) of polyclonal rabbit sera that were obtained by immunization with purified native mitogillin. Consequently, we developed an enzyme-linked immunosorbent assay for detection of IgG, IgM, and IgA antibodies to recombinant mitogillin. In serum samples of patients suffering from aspergilloma (AO; n = 32), invasive pulmonary aspergillosis (IPA; n = 42), or invasive disseminated aspergillosis (IDA; n = 40), a good correlation of production of IgG antibody against mitogillin and clinical disease was observed (for patients with AO, 100% [32 of 32] were positive; for patients with IPA, 64% [31 of 42] were positive; for patients with IDA, 60% [24 of 40] were positive). In contrast, positive titers for serum IgG and IgM antibodies against mitogillin were found in only 1.3% of the serum samples of healthy volunteers and positive titers for IgA antibody were found in only 1.0% of the serum samples of healthy volunteers (n = 307; specificity = 95.4%). These results indicate that recombinant mitogillin expressed in E. coli can be used for improvement of the serodiagnosis of A. fumigatus-associated diseases.
AuthorsM Weig, M Frosch, K Tintelnot, A Haas, U Gross, B Linsmeier, J Heesemann
JournalJournal of clinical microbiology (J Clin Microbiol) Vol. 39 Issue 5 Pg. 1721-30 (May 2001) ISSN: 0095-1137 [Print] United States
PMID11325981 (Publication Type: Evaluation Study, Journal Article)
Chemical References
  • ASPF1 protein, Aspergillus fumigatus
  • Allergens
  • Antibodies, Fungal
  • Antigens, Fungal
  • Antigens, Plant
  • Fungal Proteins
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin M
  • Recombinant Proteins
  • MITF protein, Aspergillus restrictus
  • Ribonucleases
Topics
  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Allergens
  • Animals
  • Antibodies, Fungal (blood)
  • Antigens, Fungal (immunology)
  • Antigens, Plant
  • Aspergillosis (diagnosis)
  • Aspergillus fumigatus (immunology)
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Fungal Proteins (genetics, immunology)
  • Humans
  • Immunoglobulin A (blood)
  • Immunoglobulin G (blood)
  • Immunoglobulin M (blood)
  • Male
  • Middle Aged
  • Rabbits
  • Recombinant Proteins (immunology)
  • Reproducibility of Results
  • Ribonucleases

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