<p>Experimental design used to investigate the effects of rAg85B on OVA-induced allergic lung inflammation (<i>A</i>). BALB/c mice were intraperitoneally immunized with OVA on days 0 and 14. On days 21 to 25 after the first immunization, mice were exposed to aerosolized 5% OVA for 20 min. Three hours prior to OVA inhalation, the mice were i.p. (100 µg; days 0 and 14) and i.n. (20 µg; days 21, 23, and 25) administered rAg85B. One day after the last challenge, the BAL cells were counted (<i>B</i>) and OVA-specific serum IgE concentrations were determined by ELISA (<i>C</i>). Flow cytometry of BAL cells from naïve or OVA sensitized BALB/c mice treated with PBS or rAg85B, stained with anti-Gr-1 and anti-Siglec-F. Numbers adjacent to outlined area indicate percent of eosinophils (Gr-1<sup>dull</sup>, Siglec-F<sup>+</sup>), and neutrophils (Gr-1<sup>+</sup>, Siglec-F<sup>neg</sup>) (<i>D</i>). Formalin-fixed tissue sections were stained with hematoxylin and eosin to visualized cell recruitment (upper row, scale bar, 100 mm), Masson's trichrome (center row, scale bar, 100 mm), and α-smooth muscle actin (lower row, scale bar, 50 mm). Numbers in quadrants indicate the score scale from 0 to 5 in each. (<i>E</i>). Data are representative of at least three independent experiments. (*P<0.05, **P<0.01 compared with OVA control. error bars, s.d.; n = 6 mice).</p
