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[Molecular aspects and diagnostic value of fungal allergens]
R Crameri1, S Hemmann, C Mayer
1Schweizerisches Institut für Allergie- und Asthmaforschung (SIAF), Davos, Schweiz. crameri@siaf.unizh.ch
Abstract:
Cloning, sequencing and production of highly pure recombinant allergens allows to produce perfectly standardised allergen preparations. The development of a new cloning system based on filamentous phage allowed the fast isolation and characterisation of allergens from the fungus Aspergillus fumigatus. The produced recombinant allergens were tested in serological and clinical studies as well as for their performance for routine assessments in the ImmunoCAP-system. Thereby, a perfect correlation between skin test results and serology was found showing the potential of recombinant allergens for the diagnosis of allergic diseases. Moreover, the characterisation of fungal allergens substantially contributes to our understanding of the molecular nature of proteins involved in the elicitation of allergic reactions. Apart from allergenic proteins with unknown biological function, fungal allergens can be subdivided into two classes: 1. Species-specific, secreted proteins and 2. cytoplasmic, even in phylogenetically distant organisms, well conserved proteins. These fungal allergens show extended sequence similarity, a high level of IgE cross-reactivity and in some cases also cross-reactivity with homologous human proteins indicating autoimmune reactions involved in fungal allergy.
Insights
Highly pure recombinant allergens from Aspergillus fumigatus were produced using a new cloning system. These standardized allergens show potential for diagnosing fungal allergies, correlating well with skin tests and serology.
Area of Science:
- Molecular biology
- Immunology
- Allergology
Background:
- Standardized allergen preparations are crucial for accurate allergy diagnosis.
- Recombinant allergens offer a path to highly pure and consistent diagnostic tools.
- Understanding fungal allergens is key to deciphering allergic reactions.
Purpose of the Study:
- To develop a new cloning system for rapid isolation and characterization of fungal allergens.
- To produce highly pure recombinant allergens from Aspergillus fumigatus.
- To evaluate the diagnostic performance of these recombinant allergens.
Main Methods:
- Development of a filamentous phage-based cloning system.
- Cloning, sequencing, and production of recombinant Aspergillus fumigatus allergens.
- Serological and clinical studies, including ImmunoCAP system assessments.
- Comparison of skin test results with serological data.
Main Results:
- Successful isolation and characterization of Aspergillus fumigatus allergens using the new system.
- High correlation observed between skin test results and serology for recombinant allergens.
- Demonstrated potential of recombinant allergens for routine allergy diagnosis.
- Characterization revealed two classes of fungal allergens: species-specific secreted and conserved cytoplasmic proteins.
Conclusions:
- Recombinant allergens enable perfectly standardized preparations for allergy diagnosis.
- Filamentous phage system facilitates rapid allergen isolation and characterization.
- Recombinant allergens show strong diagnostic potential, correlating with established methods.
- Fungal allergens exhibit sequence similarity, IgE cross-reactivity, and potential autoimmune links.