Figure 4

Number and maturation status of lung CD11c ⁺ CD11b ⁺ cells following NO ₂ and ova exposure. Mice were exposed to 1 hour of 15 ppm 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 expression of MHCII and the co-stimulatory molecules CD40 and CD86 48 hours post exposure (C). Maturation markers are graphed as median fluorescence intensity (MFI). 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. Differences did not reach statistical significance by unpaired Student's t test.