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Pilot project: In vitro system for characterizing the allergic potential of human and animal pathogenic fungi - Method for allergen screening of fungi

The aim of the project is to investigate whether the use of live lung sections from interleukin-33-reporter mice allows to determine the allergenicity of human and animal pathogenic fungi with zoonotic potential. Interleukin 33 is a cytokine which is involved in the induction of a T-helper cell 2 response and thus can set the course for an allergy very early in the immune response. This early interleukin 33 response to allergens is possible without prior stimulus, so that it is postulated that naive animals can be used to test their response to allergens. In these reporter mice, the gene for interleukin 33 is replaced by citrine, so that an expression of interleukin 33 can be detected by fluorescence microscopic methods by measuring the citrine signal. By transferring from the animal to the cell culture using intact organ pieces, in this case the lung, it is possible to test not only one stimulus with the help of an animal, but also several stimuli (PCLS, prescision cut lung slices) by using cuts (several cuts can be obtained from one lung). It is thus possible to test an individual's response to different stimuli in parallel and also reduce the number of animals required.

The fungus whose allergenic potential is being investigated is Cryptococcus neoformans, an encapsulated unicellular fungus that occurs ubiquitously in the environment and is spread via bird excrements, among other things. It is suspected to cause allergies in people with an intact immune system. Cryptococci are one of the few fungal groups with zoonotic potential. Infections of various domestic animal species, especially dogs and cats, have been described. These fungi pose a particular threat to immunocompromised patients. However, this aspect is not part of the pilot project. In addition to different cryptococcal strains, the allergenic components are to be identified subsequently, i.e. whether potentially allergenic components are more likely to occur in the capsule, the cell wall or in the cytoplasm. In further studies with the living lung sections, the allergenicity of fungal proteins that were associated with T-helper cell 2 responses in preliminary experiments in mouse models will be investigated. The project will be accompanied by investigations of the interaction between pneumocytes and immune cells using molecular biological and flow cytometric methods. At the end of the project a workshop is planned to introduce members of the zoonoses platform to the PCLS method.

01. September 2019 - 31. August 2020

PD Dr. Uwe Müller
Universität Leipzig
Institut für Immunologie
Deutscher Platz 5
04103 Leipzig
Tel +49 341-97 31 224
Fax +49 341-97 31 229