Identification and evaluation of a major cytotoxin of A. fumigatus
Abstract
Aspergillus fumigatus is a highly pathogenic fungus causing a wide spectrum of diseases in immunocompromised as well as immunocompetent hosts. The present work was undertaken to evaluate the cytotoxic nature of fractionated antigens of A. fumigatus against the mammalian cell lines (J774, RAW, CHO and L929). An enriched protein antigenic fraction of A. fumigatus was subjected to con A Sepharose and phenyl Sepharose chromatography. Antigenic fractions, ConAub (conA unbound) and PSC III (fraction III of phenyl Sepharose column) containing low mw antigens showed higher cytotoxicity as compared to other antigenic fractions. PSC III was further purified on HPLC resulting in an 18 kDa homogeneous protein. The purified protein showed high ELISA absorbance values for specific IgG and IgE antibodies in sera of ABPA patients. Monoclonal antibody raised against Asp fl, a major allergen/antigen of A. fumigatus recognised the purified 18 kDa by ELISA and western blot. The 18 kDa allergen/antigen or Asp fl showed similar toxicity towards all the four cell lines (macrophage and fibroblast) with an IC50 of 75 ng/ml or 4.16 nM. Reduction in toxicity of 18 kDa at low temperatures and potentiation in presence of ammonium chloride and monensin indicates mechanism of internalisation of 18 kDa in eukaryotic cells is similar to alpha-sarcin. The present work shows that the 18 kDa allergen/antigen (Asp fl) is a major cytotoxin secreted by A. fumigatus which may play multiple roles in the pathogenesis of Aspergillosis through allergenicity, antigenicity and cytotoxicity.
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Small proteins (5 to 15 kDas) rich in disulfide bridges are released in vivo in experimental models of IA.
“This is the case for small proteins (5 to 15 kDas) rich in disulfide bridges, which protect them from degradation in biological fluids (129, 130).”
From Aspergillus fumigatus and Aspergillosis in 2019 (2019) · cites this paper as [130]
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- Authors
- Madan, T., Arora, N., & Sarma, P. U.
- Affiliations
- Institute of Genomics and Integrative Biology (IN)
- Venue
- Molecular and Cellular Biochemistry
- Published
- 1997-02
- Volume
- 167
- Issue
- 1-2
- Pages
- 89-97
- ISSN
- 0300-8177 (ISSN-L)
- 1573-4919
- OpenAlex
- W50712442
- Cited by
- 21
- References
- 27
- Citations by year
- 2026: 1 · 2021: 1 · 2019: 1
- FWCI
- 2.30
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Molecular and Cellular Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Council of Scientific and Industrial Research, India
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Immunology and Microbiology › Immunology › Toxin Mechanisms and Immunotoxins
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Mycotoxins in Agriculture and Food
- Keywords
- Aspergillus fumigatus, cytotoxicity, fungal allergens, allergic bronchopulmonary aspergillosis, protein purification, monoclonal antibodies, fungal pathogenesis, immunocompromised patients
- MeSH terms
- Animals, Antigens, Fungal, Aspergillosis, Allergic Bronchopulmonary, Aspergillus fumigatus, Cells, Cultured, Chromatography, High Pressure Liquid, Cytotoxins, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Fibroblasts, Humans, Macrophages, Mammals
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