Fig. 2.

Direct fungal contact stimulates cytokine response in the organotypic bronchiole model of invasive pulmonary aspergillosis. a Experimental overview depicting how the bronchiole models are inoculated with spores, how the spores germinate and grow into hyphae, and how media is collected from the models for chemical analysis. b Photo of bronchiole model 18 h after inoculation of WT A. fumigatus spores directly into the center, epithelial cell-lined lumen. Fungal hyphae extend through the epithelial barrier into the surrounding matrix. Scale bar is 250 μm. c Closer image of fungal hyphae in b protruding from the center lumen. Scale bar is 100 μm. d Cell viability analysis of center, epithelial cell-lined lumens at 4 and 18 h post inoculation with spores. Conditions were performed in triplicate with the mean value plotted as a bar and error bars representing standard error of the mean. Data were analyzed using an ordinary two-way ANOVA with Tukey’s multiple comparisons test; none of the comparisons met the significance threshold of a p-value < 0.05. e–g Cytokine concentrations measured in media collected from the side, endothelial cell-lined lumens of bronchiole models inoculated with WT A. fumigatus spores or ΔlaeA A. fumigatus spores normalized to control, non-inoculated bronchiole models. Each plotted bar represents the mean of nine organotypic devices prepared on three separate days and error bars represent standard error of the mean. Data were analyzed using an ordinary one-way ANOVA with Tukey’s multiple comparisons test and horizontal brackets denote comparisons between conditions that are statistically significant (p-value < 0.05). Additional cytokines from this experiment are presented in Supplementary Fig. 3