Figure 5
From: Oxidative stress-mediated iNKT-cell activation is involved in COPD pathogenesis
Respiratory antigen presenting cells (APCs) isolated from cigarette smoke (CS)-exposed mice activate invariant natural killer T (iNKT) cells. (a) Twelve weeks after CS exposure, the number of alveolar macrophages (AM) (CD45+ CD11c+ F4/80+) and dendritic cell (DC; CD45+ CD11c+ F4/80−) in the lung compartment were determined. Data are representative of one experiment out of three performed (mean±s.e.m., n=6). (b) Activation status of lung DC and AM. Representative histograms of CD40 and I–A on AM and DC are shown. MFI are represented on the dot plots. (c) Expression of CD1d on AM and DC. Data are representative of one experiment out of three performed (mean±s.e.m., n=6). *P<0.05, **P<0.01. (d) Pulmonary AM, CD11b+ DC, and CD103+ DC were sorted from air- or CS-exposed mice (2 or 10weeks post-treatment), RNA samples were prepared and IL-12p40, CD1d, glucosylceramide synthase Ugcg, and GM3 synthase St3gal5 mRNA expression were measured by quantitative RT–PCR. Data are normalized to expression of Gapdh and are expressed as fold increased over average gene expression in air-exposed mice. Each group is a pool of cells from 10 mice and is representative of three independent experiments. (e) Bone marrow-dendritic cells (BM-DC) were differenciated and exposed or not to CSE overnight before coculture with sorted iNKT cells. Interleukin (IL)-17 and interferon (IFN)-γ levels were measured by ELISA in the coculture supernatants 48 h later. Data represent the mean±s.e.m. of three independent experiments performed in triplicate. *P<0.05.
