In vivo analysis of temporal and spatial disease progression and immune cell recruitment during invasive Aspergillus fumigatus infection

Project A3 aims to uncover the key players of secondary immune defense mechanisms in neutropenic and/or alveolar macrophage-depleted conditions against A. fumigatus to uncover how these cells contribute to the regulation of inflammation and fungal clearance. In the first funding period A3 analyzed in vivo pulmonary A. fumigatus disease progression and the spatiotemporal coordination of first line immune cell responses in clinically relevant immunosuppressed murine models. A3 advanced and established novel optical imaging techniques, particularly in vivo bioluminescence imaging and 3D light sheet fluorescence microscopy with optimized automated image processing and analysis. In the present funding period A3 aims to define A. fumigatus strain-specific lung cytokine response and dissect cytokine/ chemokine receptor-ligand interactions after infection. Furthermore, A3 will explore second-line immune defense mechanisms in neutropenic and/or macrophage depleted mice to understand how second line innate immune cells prime adaptive immunity. To further elucidate the molecular mechanisms involved in in vivo NET formation and the role of antifungal vesicles in NETosis, A3 will extend its imaging tool box to intravital and super-resolution microscopy to unravel A. fumigatus-host myeloid cell interactions at the cellular and molecular level.