Figure 3.

Chronic exposure to purified Aspergillus fumigatus chitin elicits inflammatory changes in the lung. C57BL/6 wild-type (WT) mice were exposed intratracheally to A. fumigatus-derived chitin as outlined in (A). Vehicle-treated mice received PBS at identical time points. Twenty-four hours after the last challenge, lung cells were isolated via bronchoalveolar lavage (BAL) were collected, homogenized, and clarified by centrifugation, enumerated, Fc-blocked, stained with a live/dead staining kit, and then stained using cell surface markers for neutrophils, eosinophils, and dendritic cells (B) or CD4 T cells (C) and quantified via flow cytometry. The figure illustrates cumulative data from two independent studies (n = 5 mice/group, per study). Each dot presents a single mouse. The middle line represents the mean. Data is expressed as absolute cell number in bronchoalveolar lavage fluid (BALF). *,**,***P value of <0.05, <0.01, and <0.0001, respectively (two-tailed Student’s t test). Twenty-four hours after last challenge, the left lungs were collected, homogenized and clarified by centrifugation. D: IL-33, CCL17, and CCL22 levels were quantified by ELISA. IL-4, IL-5, and IL-13 levels (E) and IFN-γ and IL-17A (F) were quantified by Milliplex. The figure illustrates cumulative data from three independent studies (n = 4 or 5 mice per group, per study). Each dot presents a single mouse. The middle line represents the mean. Data are expressed as pg/mL in lung homogenate. * and *** represent a P value of < 0.05 and < 0.0001, respectively (two-tailed Student’s t test). [Image created with BioRender.com.]