Figure 6. Adoptive transfer of HDM-pulsed, CPT or curdlan-treated cDC2 induces a Th17 bias and neutrophil infiltration in WT recipients.

(A) Schematic of adoptive transfer protocol. WT cDC2s were incubated with HDM (50 μg/ml) in the presence of CPT or curdlan prior to i.n. transfer to WT mice (B6 mice, 1 × 10⁶ cells/recipient) on day 0 and 14. HDM (12.5 µg/mouse) was used for the i.n. challenge on days 19, 20 and 21. On day 22, lungs from each group were harvested and processed a single cell suspension. (B) IL-17A level in the HDM (50 μg/ml)-stimulated lung cells. BAL fluid was analyzed for (C) IL-17 level (ELISA). (D) Total cells (E) neutrophils, (F) macrophages and (G) eosinophils number were counted in the BAL fluid. (H) Relative expression of Neut (Cxcl2 and Cxcl3)- and/Eos (Cd24a and Dpp4)- infiltration related genes in the lung tissue. Expression of each gene was normalized by expression of epithelial specific housekeeping gene, Epcam. (I) Airway resistance after MCh challenge was measured in the various experimental groups. Data were collected from four animals in each group and are representative of two independent experiments; *p<0.05, **p<0.01, ***p<0.001 by two-tailed Student’s t-test.

Figure 6—figure supplement 1. Adoptive transfer of OVA-pulsed, CPT or curdlan-treated cDC2s induces a Th17 bias and neutrophilic asthma in WT recipients.

(A) Schematic of adoptive transfer protocol. WT cDC2s were incubated with OVA (200 μg/ml) in the presence of CPT or curdlan prior to i.n. transfer to WT (B6 mice) or IL-17-GFP mice (1 × 10⁶ cells/recipient) on day 0 and 14. OVA (50 µg/mouse) was used for the i.n. challenge on days 19, 20 and 21. (B) Total cell number, (C) neutrophils, (D) macrophages and (E) eosinophils number of BAL fluid. IL-17A levels from the (F) BAL fluid and (G) OVA (200 μg/ml)-stimulated lung cells. (H) Isolated lung cells from IL-17-GFP mice were analyzed (FACS) for the eGFP fluorescence intensity after PMA/ionomycin stimulation. Data were collected from five animals in each group and are representative of two independent experiments; *p<0.05, **p<0.01, ***p<0.001 by two-tailed Student’s t-test.