Proteomic characterisation of <i>Aspergillus fumigatus</i> – host interactions using the <i>ex-vivo</i> pig lung (EVPL) model<b>.</b>Suppfigures and tables evpl paper.pdf

<i>Aspergillus fumigatus</i> is an opportunistic fungal pathogen of the human airway that can cause a variety of chronic infections, typically in the context of pre-existing lung damage. The interaction of <i>A. fumigatus</i> with <i>ex-vivo</i> pig lung (EVPL) samples was characterized at the proteomic level to provide insights into how the fungus may interact with pulmonary tissue <i>in vivo</i>. This model has many advantages, because pigs share 90% immunological homology with humans and display many anatomical similarities. EVPL also retains resident immune cells, has richer cellular complexity compared to <i>in-vitro</i> models, and has a microbiome. Label-free quantitative proteomic analysis identified the metabolism and development of <i>A. fumigatus</i> on the EVPL alveolar sections; at 48 h, there was an increased abundance of proteins associated with carbon metabolism (e.g., malate dehydrogenase (+ 8.2 fold increase)), and amino acid metabolism and biosynthesis (e.g., 5-methyltetrahydropteroyltriglutamate--homocysteine S-methyltransferase, (+5.04 fold)) at 72 h. Porcine tissue remained responsive to the pathogen with proteins that increased in abundance associated with innate immune recruitment (e.g., protein S100-A8 (+28.5 fold) and protein S100-A9 (calgranulin-B) (+7.25 fold)) at 24 h, while proteins associated with neutrophil degranulation (e.g., elastase, neutrophil (-2.74 fold)) decreased in abundance. At 96 h, the infected tissue demonstrated enhanced expression of fibrotic markers (e.g., fibrillin 1, collagen type IV alpha 1 chain, and alpha 2 chain, increased by +16.44, +15.42 and +11.95 fold respectively). These results validate the use of this model for studying pathogen-host interactions and highlight how <i>A. fumigatus</i> interacts with pulmonary tissues during colonization.