TY  - JOUR
A1  - Yu, Yidong
A1  - Wolf, Ann-Katrin
A1  - Thusek, Sina
A1  - Heinekamp, Thorsten
A1  - Bromley, Michael
A1  - Krappmann, Sven
A1  - Terpitz, Ulrich
A1  - Voigt, Kerstin
A1  - Brakhage, Axel A.
A1  - Beilhack, Andreas
T1  - Direct Visualization of Fungal Burden in Filamentous Fungus-Infected Silkworms
JF  - Journal of Fungi
N2  - Invasive fungal infections (IFIs) are difficult to diagnose and to treat and, despite several available antifungal drugs, cause high mortality rates. In the past decades, the incidence of IFIs has continuously increased. More recently, SARS-CoV-2-associated lethal IFIs have been reported worldwide in critically ill patients. Combating IFIs requires a more profound understanding of fungal pathogenicity to facilitate the development of novel antifungal strategies. Animal models are indispensable for studying fungal infections and to develop new antifungals. However, using mammalian animal models faces various hurdles including ethical issues and high costs, which makes large-scale infection experiments extremely challenging. To overcome these limitations, we optimized an invertebrate model and introduced a simple calcofluor white (CW) staining protocol to macroscopically and microscopically monitor disease progression in silkworms (Bombyx mori) infected with the human pathogenic filamentous fungi Aspergillus fumigatus and Lichtheimia corymbifera. This advanced silkworm A. fumigatus infection model could validate knockout mutants with either attenuated, strongly attenuated or unchanged virulence. Finally, CW staining allowed us to efficiently visualize antifungal treatment outcomes in infected silkworms. Conclusively, we here present a powerful animal model combined with a straightforward staining protocol to expedite large-scale in vivo research of fungal pathogenicity and to investigate novel antifungal candidates.
KW  - fungal infection model
KW  - calcofluor white staining
KW  - Aspergillus
KW  - Lichtheimia
KW  - silkworm
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228855
SN  - 2309-608X
VL  - 7
IS  - 2
ER  - 
TY  - JOUR
A1  - Trinks, Nora
A1  - Reinhard, Sebastian
A1  - Drobny, Matthias
A1  - Heilig, Linda
A1  - Löffler, Jürgen
A1  - Sauer, Markus
A1  - Terpitz, Ulrich
T1  - Subdiffraction-resolution fluorescence imaging of immunological synapse formation between NK cells and A. fumigatus by expansion microscopy
JF  - Communications Biology
N2  - Expansion microscopy (ExM) enables super-resolution fluorescence imaging on standard microscopes by physical expansion of the sample. However, the investigation of interactions between different organisms such as mammalian and fungal cells by ExM remains challenging because different cell types require different expansion protocols to ensure identical, ideally isotropic expansion of both partners. Here, we introduce an ExM method that enables super-resolved visualization of the interaction between NK cells and Aspergillus fumigatus hyphae. 4-fold expansion in combination with confocal fluorescence imaging allows us to resolve details of cytoskeleton rearrangement as well as NK cells' lytic granules triggered by contact with an RFP-expressing A. fumigatus strain. In particular, subdiffraction-resolution images show polarized degranulation upon contact formation and the presence of LAMP1 surrounding perforin at the NK cell-surface post degranulation. Our data demonstrate that optimized ExM protocols enable the investigation of immunological synapse formation between two different species with so far unmatched spatial resolution.
KW  - biological fluorescence
KW  - fluorescence imaging
KW  - imaging the immune system
KW  - infectious diseases
KW  - super-resolution microscopy
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-264996
VL  - 4
IS  - 1
ER  -