Figure 2.
A: Effect of cytokines on development of steroid resistance. PBMCs from allergic asthmatic patients were cultured with a combination of ILC2-stimulating cytokines in the presence or absence of Dex for 5 days. The cells were then stained for FCM and gated for IL5+ ILC2s as described under Figure 1A. *P<0.05 (n=6). B: Kinetics of development of resistance against inhibition of IL5+ ILC2s by Dex. PBMCs were cultured with IL2/TSLP plus vehicle or Dex for 1, 3 and 5 days and then analyzed for IL5+ ILC2s by FCM. *P<0.05 (n=4). C&D: Lin- cells (0.4 × 106 per culture) from peripheral blood obtained from 3 allergic rhinitis (non-asthmatic) patients were cultured with IL2/IL33 or IL2/TSLP with and without Dex (10-7 M) for 5 days. Cell-free supernatant was analyzed for IL5 by ELISA. Cells were analyzed by FCM for intracellular IL5. Dex inhibition of IL5 secretion and intracellular IL5 expression are presented in panel C and D respectively. P value for data on panel C was calculated by paired t test due to low N. E: Comparison of the effect of Dex on IL5+ ILCs (lin-CRTH2+IL7Rα+) and IL5+ lineage+ cells in BAL. BAL cells were cultured with vehicle or Dex for 5 days, stained and first gated for CD45+ cells and then for lin-, CRTH2+, IL7Rα+, IL5+ cells (IL5+ ILCs) and lin+ IL5+ cells. Each data point is an individual asthma patient (n=9).