Figure 5.

Determination of in vivo leukocyte and intracellular cytokine production. Freshly harvested A. fumigatus conidia (CEA10, 12 × 10⁹) were delivered via aerosolization to immunosuppressed C57BL/6 mice. Mice in the steroid model were immunosuppressed via subcutaneous injection of cortisone acetate, while mice in the chemotherapeutic model were immunosuppressed via subcutaneous injection of cortisone acetate and intraperitoneal injection of cyclophosphamide three days prior to inoculation. Leukocyte populations and counts were determined from (a) bronchoalveolar lavage fluid (BALF) and (b–f) lung tissue. Cell counts were determined for (c) CD4+, CD8+, and NK cell populations and (d) for positive intracellular staining of IL-17, IL-4, or IFN-γ. Cells counts were also determined (e) CD103+, monocytoid (mDC), plasmocytoid (pDC), and conventional (cDC) dendritic cells and for (f) positive intracellular staining of IL-12 and IL-4. Statistical significance was determined use the student t-test. Statistical significance (p < 0.05) between the two models for a given cell type is indicated by an asterisk. Statistical significance via Duncan’s multiple range test (p < 0.05) for CD4+ T cells stained positive IL-4, IL-17, and IFN-γ is indicated with ‘A’ (steroid model). Statistical significance via Duncan’s multiple range test (p < 0.05) for NK cells stained positive IL-4, IL-17, and IFN-γ is indicated with ‘B’ (steroid model). Statistical significance (p < 0.05) between a given dendritic cell population for count of cells positive for IL-12 versus IL-4 production is indicated with an ‘C’ in the steroid model, and a ‘D’ for the chemotherapeutic model. All experiments were run in independent replicates, n = 8.