Article Figures & Data
Figures
- Fig. 1.
Light microscopy of typical A. fumigatussporulating structures.
- 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.
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.
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.
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.
Role of the host innate immunity against A. fumigatus and its modulation by immunosuppressive agents used in BMT.
- Fig. 7.
Cytokines and their possible role in modulating cellular immunity in response to A. fumigatus in experimental murine aspergillosis.
Tables
- Table 1.
Selected features used to classify A. fumigatus to the species and strain levels
Feature Reference(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-PAGEf Localizationb Biochemical function Gene cloned Recombinant protein Reference(s) 12 ? ? + + 123 18c S RNase + + 21,126, 168, 343, 344, 349, 377, 424, 433, 434 19 IC Peroxisomal protein + + 123 19c(67)d S Superoxide dismutase − NAe 236, 267,268 20 S ? − NA 578 27 IC Superoxide dismutase + + 123, 125,256 28 S ? − NA 132 30 ? ? + + 123 33 S Serine 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 38 S Aspartic protease + + 424,521-523 40 S Metalloprotease + + 286, 424, 426,607 82 IC Metalloprotease + − 277 88c S Dipeptidyl peptidase + + 41, 240, 313 90c(350)d S Catalase + + 89,252, 382 93 IC ? − NA 243 94 S Dipeptidyl 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
Antigen Biological fluid Detection limit (ng/ml) Reference(s) Galactofuran-containing antigensa Serum, urine, BAL fluid 0.5–1 16,74, 557, 558, 628, 630, 638, 683, 684, 690 29, 18, 11 kDab Urine ?c 241 β(1-3)glucan Serum 10−2 421, 468,469, 744 - Table 4.
Reagents and assays used in the detection of galactofuran-containing antigens in serum of patient with IA
Assay Detection 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 antibody 2 205 MAb 1 628 Method of assay Latex 15 178, 242, 395, 690 Radioimmunoassay 7 639, 721 ELISA inhibition 5 345, 496, 573 ELISA sandwich 1 628 Ultrasound + latex + videomicroscopy 0.1 232 ↵a TCA, trichloroacetic acid; PBS, phosphate-buffered saline.
- Table 5.
Analysis of PCR results in patients with and without IA
Specimen Probe Total no. of patients No. positive Reference PCR+/IA+a PCR+/IA−b Urine 18-kDa ribotoxin 13c 1/1 1/12 519 BAL fluid rDNA 10d 3/3 2/7 617 BAL fluid 33-kDa alkaline protease 51 5/5 6/46 643 BAL fluid rRNA 25 6/6 0/19 407 BAL fluid rRNA 70 NAh 11/70 688 BAL fluid mtDNA 52 3/3 12/49 73 Serum mtDNA 23e 5/5 0/18 72 Serum rRNA 40f 14/20 0/20 733 Plasma rRNA 77 13/13 0/64 184 Plasma rRNA 8g 3/3 0/5 672 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
Category Role in vivo Molecule Reference(s) Adhesins Promotion 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 Pigments Inhibition of phagocytosis of conidia Dihydroxynaphthalene-melanin 284, 350, 666,667 Toxic molecules Host cell death RNase (18 kDa) 19, 189, 343, 344 Erythrocyte lysis Hemolysin (30 kDa) 179-181, 206, 207,737-740 Immunosuppression Secondary metabolites, e.g., gliotoxin 10, 129, 182, 632, 633,709 Enzymes Promotion 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 phagocytosis Catalases (350 kDa and unknown) 89 Superoxide dismutases (27 and 67 kDa) 125, 267, 268 Epithelial damage Phospholipase(s) 57
- 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
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