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Aspergillus fumigatus and Aspergillosis

Jean-Paul Latgé
Jean-Paul Latgé
Laboratoire des Aspergillus, Institut Pasteur, 75015 Paris, France
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DOI: 10.1128/CMR.12.2.310
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  • Fig. 1.
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    Fig. 1.

    Light microscopy of typical A. fumigatussporulating structures.

  • Fig. 2.
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    Fig. 2.

    Schematic representation of the λ3.9 probe of A. fumigatus used for molecular studies and typical hybridization patterns obtained with EcoRI-digested total DNA probed with the entire SalI-SalI fragment (A) andEcoRI fragments of the repeated sequence (B to E). The repeated element Afut1 (squares) is an inactive retroelement of 6.9 kb bounded by two LTRs (▸) and with sequences homologous to reverse transcriptase (RT), RNase H, and endonuclease (endo) encoded by the pol genes of retrotransposons.

  • Fig. 3.
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    Fig. 3.

    Schematic representation of steps involved in the development of a diagnostic test for the detection of antigen in the biological fluids of patients with IA, using GM as an example.

  • Fig. 4.
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    Fig. 4.

    Strategies currently available to disrupt genes inA. fumigatus (A) and to create a bank of A. fumigatus mutants by using the restriction enzyme-mediated integration strategy (B). E1 and E2 are endonucleases favoring a single-copy integration of the marker in the A. fumigatusgenome. wt, wild type; OMP, orotidine-5′-phosphate; 5FOA, 5-fluoroorotic acid.

  • Fig. 5.
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    Fig. 5.

    Effect of different immunosuppressive drugs and routes of infection on the number of conidia required to induce IA in outbred Swiss mice. Data from references 94, 110, 111, 116, 171, 224,264, 423, 540, 581, 612, 614, 641, and649.

  • Fig. 6.
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    Fig. 6.

    Role of the host innate immunity against A. fumigatus and its modulation by immunosuppressive agents used in BMT.

  • Fig. 7.
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    Fig. 7.

    Cytokines and their possible role in modulating cellular immunity in response to A. fumigatus in experimental murine aspergillosis.

Tables

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  • Table 1.

    Selected features used to classify A. fumigatus to the species and strain levels

    FeatureReference(s)
    Species characterizationa
     Culture and morphology 341, 518, 577,595
     Secondary metabolites 200
     Specific unique and repeated DNA sequencesb 105, 211, 218, 502,582
     DNA-DNA reassociation 501
    Strain characterization
     Microsatellite patterns 34
     Hybridization patterns of endonuclease-digested DNA with AFUT1 133,217
    • ↵a Isoenzyme patterns (369, 400, 543,554), ethidium-bromide visualized RFLPs (82, 144), and IGS probes (514, 618) can be used to rank strains at a subspecies level.

    • ↵b Used directly for comparative analysis of sequence data obtained with unique nuclear (211) or ITS1 and ITS2 ribosomal (105, 210, 502) genes or indirectly in Southern hybridization experiments (218, 582).

  • Table 2.

    Purified antigenic proteinsa ofA. fumigatus reported in the literature

    Mol mass (kDa) from SDS-PAGEfLocalizationbBiochemical functionGene clonedRecombinant proteinReference(s)
    12??++ 123
    18c SRNase++ 21,126, 168, 343, 344, 349, 377, 424, 433, 434
    19ICPeroxisomal protein++ 123
    19c(67)d SSuperoxide dismutase−NAe 236, 267,268
    20S?−NA 578
    27ICSuperoxide dismutase++ 123, 125,256
    28S?−NA 132
    30??++ 123
    33SSerine protease++ 287, 424, 428,435, 436, 520
    34??++ 123
    36 (70?)d S?++ 26, 27,87, 88, 90, 197, 239, 333, 338, 380, 504, 648
    38SAspartic protease++ 424,521-523
    40SMetalloprotease++ 286, 424, 426,607
    82ICMetalloprotease+− 277
    88c SDipeptidyl peptidase++ 41, 240, 313
    90c(350)d SCatalase++ 89,252, 382
    93IC?−NA 243
    94SDipeptidyl peptidase++ 42
    • ↵a Antigens reacting with antibodies from immunocompetent patients with aspergillosis have been purified by biochemical or molecular biology strategies.

    • ↵b S, secreted (including possible cell wall localization); IC, intracellular.

    • ↵c Most discriminant antigens.

    • ↵d The molecular mass of the native protein is indicated in parentheses.

    • ↵e NA, not applicable.

    • ↵f SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

  • Table 3.

    Molecules detected in biological fluids of patients with IA due to A. fumigatus

    AntigenBiological fluidDetection limit (ng/ml)Reference(s)
    Galactofuran-containing antigensa Serum, urine, BAL fluid0.5–1 16,74, 557, 558, 628, 630, 638, 683, 684, 690
    29, 18, 11 kDab Urine?c 241
    β(1-3)glucanSerum10−2 421, 468,469, 744
    • ↵a Glycoprotein and polysaccharide.

    • ↵b Plus other minor antigens.

    • ↵c Unknown; detection by immunoblotting.

  • Table 4.

    Reagents and assays used in the detection of galactofuran-containing antigens in serum of patient with IA

    AssayDetection limit (ng/ml)Reference(s)
    Pretreatment of seruma
     Perchloric acid, room temperature 603
     Citric acid, boiling 12, 294, 639,719-721
     TCA, boiling 496,573
     PBS, boiling 177, 557,728
     EDTA, boiling 178, 205, 389, 395,628
    Type of Detector Antibody
     Polyclonal antibody2 205
     MAb1 628
    Method of assay
     Latex15 178, 242, 395, 690
     Radioimmunoassay7 639, 721
     ELISA inhibition5 345, 496, 573
     ELISA sandwich1 628
     Ultrasound + latex + videomicroscopy0.1 232
    • ↵a TCA, trichloroacetic acid; PBS, phosphate-buffered saline.

  • Table 5.

    Analysis of PCR results in patients with and without IA

    SpecimenProbeTotal no. of patientsNo. positiveReference
    PCR+/IA+aPCR+/IA−b
    Urine18-kDa ribotoxin13c 1/11/12 519
    BAL fluidrDNA10d 3/32/7 617
    BAL fluid33-kDa alkaline protease515/56/46 643
    BAL fluidrRNA256/60/19 407
    BAL fluidrRNA70NAh 11/70 688
    BAL fluidmtDNA523/312/49 73
    SerummtDNA23e 5/50/18 72
    SerumrRNA40f 14/200/20 733
    PlasmarRNA7713/130/64 184
    PlasmarRNA8g 3/30/5 672
    • a Proven or highly probable disease based on clinical data.

    • b Healthy donor or patient without any clinical signs of IA.

    • ↵c Another urine sample was PCR positive, but no clinical data were given for the patient.

    • ↵d Thirteen other patients were analyzed, but clinical data were insufficient to classify.

    • ↵e Nineteen patients were positive for GM.

    • ↵f Twelve patients were positive for GM by the latex agglutination test.

    • ↵g A total of 3 of 189 blood samples from 103 control patients gave false-positive results.

    • ↵h NA, not applicable.

  • Table 6.

    A. fumigatus molecules with a putative role in virulence

    CategoryRole in vivoMoleculeReference(s)
    AdhesinsPromotion of interactions of host proteins and cells with A. fumigatus Complement receptor (54–58 kDa) 626
    Laminin receptor (72 kDa) 664
    Hydrophobins (14 and 16 kDa) 484, 649
    PigmentsInhibition of phagocytosis of conidiaDihydroxynaphthalene-melanin 284, 350, 666,667
    Toxic moleculesHost cell deathRNase (18 kDa) 19, 189, 343, 344
    Erythrocyte lysisHemolysin (30 kDa) 179-181, 206, 207,737-740
    ImmunosuppressionSecondary metabolites, e.g., gliotoxin 10, 129, 182, 632, 633,709
    EnzymesPromotion of lung matrix colonization and/or degradation of humoral factors Serine protease (33 kDa) 427, 428, 641
    Aspartic protease (38 kDa) 521, 522
    Metalloprotease (40 kDa) 286, 426
    Dipeptidylpeptidases (88 and 94 kDa) 41,42
    Antioxidants during phagocytosisCatalases (350 kDa and unknown) 89
    Superoxide dismutases (27 and 67 kDa) 125, 267, 268
    Epithelial damagePhospholipase(s) 57
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Aspergillus fumigatus and Aspergillosis
Jean-Paul Latgé
Clinical Microbiology Reviews Apr 1999, 12 (2) 310-350; DOI: 10.1128/CMR.12.2.310

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Aspergillus fumigatus and Aspergillosis
Jean-Paul Latgé
Clinical Microbiology Reviews Apr 1999, 12 (2) 310-350; DOI: 10.1128/CMR.12.2.310
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  • Top
  • Article
    • SUMMARY
    • TAXONOMY OF A. FUMIGATUS
    • CLINICAL SYMPTOMS AND DIAGNOSIS OF RESPIRATORY ASPERGILLOSIS
    • ANTIGENS AND LABORATORY DIAGNOSIS
    • ARE THERE VIRULENCE FACTORS IN A. FUMIGATUS?
    • HOST DEFENSE MECHANISMS AGAINST A. FUMIGATUS
    • MOLECULAR EPIDEMIOLOGY AND PROPHYLAXIS OF INVASIVE ASPERGILLOGIS
    • TREATMENT OF ASPERGILLOSIS
    • CONCLUSION
    • ACKNOWLEDGMENTS
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

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