Figure 6
From: Oxidative stress-mediated iNKT-cell activation is involved in COPD pathogenesis
Human invariant natural killer T (iNKT) cells are activated by cigarette smoke (CS)-exposed human dendritic cell (huDC) and human airway epithelial cell (huAEC). (a) Human monocyte-derived DC (left column) or lung epithelial cells (right column) were exposed, or not, to CSE for 24 h. After washes, cells were cocultured with primary CD3+ 6B11+ iNKT cells. Interleukin (IL)-17 and interferon (IFN)-γ were measured by ELISA. Data represent the mean±s.e.m. of three independent experiments performed in triplicate. *P<0.05, **P<0.01. (b) DC and AEC were collected 4 and 24 h after CSE exposure, RNA extracts were prepared, and St3gal5 synthase mRNA copy numbers were measured by quantitative RT–PCR. Data are normalized to expression of β-actin and are expressed as fold increased over average gene expression in air-exposed cells. (c) DC and AEC were pretreated with NAC (0.5 mM) for 1 h prior CSE exposure for 24 h. After washes, cells were cocultured with primary iNKT cells. IL-17 and IFN-γ were measured by ELISA. Data represent the mean±s.e.m. of three independent experiments performed in triplicate. *P<0.05, **P<0.01. (d) DC and AEC were treated with NAC for 1 h, and then exposed to CSE for 24 h. RNA extracts were prepared, and HO-1 and Ugcg mRNA copy numbers were measured by quantitative RT–PCR. Data are normalized to expression of β-actin and are expressed as fold increased over average gene expression in air-exposed cells. *P<0.05, **P<0.01.
