Figure 5
Role of langerin+ and CD11b+ dendritic cells (DCs) in postasthmatic mice in the steady state and upon lipopolysaccharide (LPS) exposure. (a) Postasthmatic langerin-GFP mice were exposed to LPS or phosphate-buffered saline (PBS) and lung cells were prepared 3 days later. Lung macrophages (LMs), CD11b+ LMs, CD11b+ DCs, and langerin+ (GFP+) DCs were sorted and 105 cells were incubated for 7 days with 105 carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled Thy1.1+/+ WT15 CD4+ T cells. (a) Data show representative fluorescence-activated cell sorting (FACS) profiles after gating on Thy1.1+ CD4+ T cells. (b) Cellular supernatants were analyzed for interferon-γ (IFN-γ), interleukin (IL)-5, and IL-13 content by Cytometric Bead Array (CBA). Data show mean values of triplicates±s.d. of one representative experiment (out of two). (c, d) Langerin-DTR mice were sensitized and challenged with LACK (Leishmania analog of the receptors of activated C kinase), treated with PBS or diphtheria toxin (DT) intranasally (i.n.) 16 days later. Mice were injected with CFSE-labeled WT15 Thy1.1+/+ CD4+ T cells 3 days later, and further exposed to PBS or LPS. (c) Mediastinal lymph node (MLN) and (d) blood cells were analyzed by FACS 72 h following LPS or PBS exposure. (c) Data show representative FACS profiles after gating on Thy1.1+ CD4+ T cells. The numbers indicate the frequency of cells that have undergone ⩾5 divisions. Data are expressed as mean±s.e.m.; n=7 mice per group. Three independent experiments were performed. (d) Blood cells were depleted of erythrocytes. The bars indicate the frequency of eosinophils in DT- and PBS-treated mice upon exposure to PBS or LPS. Data are expressed as mean±s.e.m.; n=7 mice per group pooled from two experiments.
