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Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions
Published on: May 16, 2020
[Preparation and Biological Characterization of Limulus Factor G-activating Substance of Aspergillus spp.]
Yuichiro Kurone1,2, Ken-Ichi Ishibashi1, Daisuke Yamanaka1
1Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences.
Abstract:
Aspergillus is a medically important fungal genus that causes a life-threatening infection known as aspergillosis in immunocompromised patients. β-1,3-Glucan is detected in the plasma of patients with aspergillosis and appears to be useful for the diagnosis of aspergillosis. In this study, we cultured Aspergillus spp. in a chemically defined liquid medium and prepared an Aspergillus water-soluble fraction (ASWS) from the culture supernatants. ASWS was found to be primarily composed of polysaccharides and proteins. Nuclear magnetic resonance analysis suggested that ASWS is a complex carbohydrate, consisting of α-1,3-glucan, β-1,3-glucan, galactomannan, and protein. The ASWS from Aspergillus fumigatus showed limulus factor G activity, whereas zymolyase-treated ASWS did not. ASWS was eliminated from the blood more rapidly than Aspergillus solubilized cell wall β-glucan. We analyzed the reactivity of human immunoglobulin towards ASWS by an enzyme-linked immunosorbent assay. Anti-ASWS antibodies were detected in human sera, with titers differing among individuals. This study demonstrated that the ASWS corresponds to the limulus factor G-activating substance found in the blood of patients with aspergillosis.
Insights
This study identifies an Aspergillus water-soluble fraction (ASWS) as a key component in diagnosing aspergillosis. ASWS, a complex carbohydrate, activates limulus factor G and is detected in patients with this serious fungal infection.
Area of Science:
- Medical Mycology
- Immunology
Background:
- Aspergillus species cause life-threatening aspergillosis in immunocompromised individuals.
- β-1,3-Glucan in plasma aids aspergillosis diagnosis.
- A limulus factor G-activating substance is found in patients with aspergillosis.
Purpose of the Study:
- To characterize the Aspergillus water-soluble fraction (ASWS).
- To investigate the diagnostic potential of ASWS in aspergillosis.
Main Methods:
- Culturing Aspergillus spp. and preparing ASWS.
- Nuclear magnetic resonance (NMR) for ASWS composition analysis.
- Limulus factor G assay and enzyme-linked immunosorbent assay (ELISA) for antibody detection.
Main Results:
- ASWS is a complex carbohydrate containing α-1,3-glucan, β-1,3-glucan, galactomannan, and protein.
- ASWS from Aspergillus fumigatus exhibited limulus factor G activity.
- Anti-ASWS antibodies were detected in human sera, indicating immune response.
Conclusions:
- ASWS is the limulus factor G-activating substance implicated in aspergillosis diagnosis.
- ASWS shows potential as a biomarker for invasive fungal infections.
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