Aspergillus fumigatus is an opportunistic fungal pathogen of the human airway that can causea variety of chronic infections, typically in the context of pre-existing lung damage. The interac-tion of A. fumigatus with ex-vivo pig lung (EVPL) samples was characterized at the proteomic levelto provide insights into how the fungus may interact with pulmonary tissue in vivo. This modelhas many advantages, because pigs share 90% immunological homology with humans anddisplay many anatomical similarities. EVPL also retains resident immune cells, has richer cellularcomplexity compared to in-vitro models, and has a microbiome. Label-free quantitative proteomicanalysis identified the metabolism and development of A. fumigatus 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 abundanceassociated 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 demon-strated enhanced abundance of fibrotic markers (e.g. fibrillin 1, collagen type IV alpha 1 chain, andalpha 2 chain, increased by + 16.44, +15.42 and + 11.95 fold, respectively). These results validatethe use of this model for studying pathogen-host interactions and highlight how A. fumigatusinteracts with pulmonary tissue during colonization.