Figure 1

Increased HIF expression enhances allergic airway inflammation and CCL2 levels in OVA sensitized/challenged mice. A. Experimental Protocol. Balb/c mice were sensitized with OVA in alum, i.p., on days 0 and 5, followed by a challenge with OVA, i.t., on days 12, 22 and 32. Mice were treated with 100 mg/kg EDHB, i.p., 24 prior to each challenge. Mice were sacrificed 4 days after the last challenge, on day 36. B. Representative H&E stained sections of lung tissues showing blood vessels (a-d) or bronchioles (e-h) obtained from OVA-sensitized/challenged (OVA) (c and g) and OVA sensitized/challenged mice treated with EDHB (OVA/EDHB) (d and h) or control mice treated with saline (SS) (a and e) and SS/EDHB (b and f) are presented in the upper panel. Representative PAS stained sections of lung tissue from OVA-sensitized mice (k) and OVA/EDHB (l), SS (i) or SS/EDHB (j) are also presented in the top panel. Quantification of the inflammatory response and mucus production for five mice from each experimental group is presented in the bottom panel. The degree of inflammatory infiltration in venules and middle size veins 100–200 μm in diameter and in bronchioles 150–300 μm in diameter from five mice from each experimental group was determined as described in “Materials and Methods.” Results are expressed as means and standard deviations of infiltrate area. (empty) saline control; (light grey) EDHB; (grey) OVA; (solid) OVA/EDHB. *p = 0.01 OVA vs. OVA/EDHB mice. C. Immunohistochemical staining of HIF-1α (a-d) and CCL2 (e-h) expression in the lung tissue of representative mice treated with OVA (c and g), OVA/EDHB (d and h), SS (a and e) and SS/EDHB (b and f) is shown in the upper panel. Quantification of HIF-1α and CCL2 immunostaining in the vessels and bronchioles of five mice from each experimental group is presented in the lower panel. Five lung sections were analyzed for each experimental group. *p = 0.001 OVA vs. OVA/EDHB mice. D. Quantification of CCL2 in BALF of mice treated with saline (empty bar), SS/EDHB (light grey), OVA (grey) and OVA/EDHB (solid bar). *p = 0.002 OVA vs. OVA/EDHB mice.