Figure 3.

Intact degranulation of IL-9R^−/− BMMC in response to Ab-mediated stimulation. BALB/c WT and IL-9R^−/− BMMC were either sensitized with 0.2 µg/mL DNP-specific IgE (clone SPE-7) or 1:100 dilutions of naïve sera (ns) or S. ratti-specific immune sera (is; taken from day 21 re-infected BALB/c mice) or left untreated overnight. BMMC were washed in PBS and 2 × 10⁵ cells were either incubated with the crosslinking model antigen DNP-HSA (IgE/DNP, 20 ng/mL), or 100 µg/mL S. ratti L3 lysate (Sr L3), or 15 µg/mL S. ratti adult lysate (Sr adult) in round bottom 96-well-plates for 20 minutes at 37 °C. Negative control BMMCs were left unstimulated (control) and positive control BMMC were activated with 100 ng/mL PMA/Ionomycin (PMA + IO). Cells were stained for FcεRIα, CD107a and CD117 expression and analyzed on a BD Calibur Flow cytometer. (a) Representative dot plots showing CD117/CD107a expression on BMMCs. Numbers show frequency of degranulated BMMC identified as CD107a⁺ cells in response to indicated stimuli. (b) Graphs show combined results out of 2 independent experiments (n = 9–10 per group) presented as mean and error bars indicate SEM.