Functional response to ozone is dependent on Toll-like receptor 4 (TLR4). Mice were exposed to 300 ppb ozone (O3) for 72 hours. (A) Airway injury was determined by level of protein in the bronchoalveolar lavage (BAL) fluid in the C3H/OuJ (tlr4-sufficient) and the C3H/HeJ (tlr4-deficient) over time. Statistical comparison: air versus O3 treatment groups (*p < 0.05); C3H/HeJ versus C3H/HeOuJ (†p < 0.05). (B) After completion of exposure to O3, physiologic response to intravenous methacholine in wild-type and tlr4 knockout mice were evaluated by direct measurements of tracheal pressures. Increased airway pressure–time index (APTI) values in O3 exposed C57BL/6 animals when compared with tlr4−/− were seen at all doses of intravenous methacholine, including 25 μg/ml (*p < 0.01), 100 μg/ml (*p = 0.02), 250 μg/ml (*p = 0.001). (B) TLR4+/+ O3, upper solid line, solid squares; TLR4−/− O3, dotted line, open triangles; TLR4+/+ air, lower solid line, solid squares; TLR4−/− air, dotted line, open diamonds. (Adapted by permission from References 24 and 86.)