Figure 3

Number and maturation status of lung CD11c ⁺ CD11b ^- cells following NO ₂ and ova exposure. Mice were exposed to 1 hour of 15ppm NO₂ or air followed by 30 minutes of aerosolized 3.4% ova. Lungs were harvested 2 and 48 hours later, single cell suspensions were generated and stained with antibodies, and cells were analyzed by flow cytometry. Dead cells were excluded by FSC and SSC gating and then total lung cells were gated comparing CD11c and CD11b expression, with future analyses focusing on the CD11c⁺CD11b^- population (bold gate, A). The total number of CD11c⁺CD11b^- cells within the lung was enumerated (B) and the maturation status of these cells was assessed by median fluorescence intensity (MFI) of MHCII and the co-stimulatory molecules CD40 and OX40L 2 hours post exposure (C). NO₂-exposed animals are represented by a thick line or black bar while air-exposed animals are represented by a thin line or white bar. Grey, filled histograms are isotype controls. Data shown are mean ± SEM with 3 animals per group and are representative of experiments performed twice. * denotes p < 0.05 and ** denotes p < 0.01 versus air control at the same time point by unpaired Student's t test. ^ denotes p < 0.05 and ^^ denotes p < 0.01 versus 2 hours post NO₂ exposure by unpaired Student's t test.