A total of forty-eight 10–12 week old mice (8 male and 8 female hTG SP-A1mice, 8 male and 8 female hTG SP-A2 mice, and 8 male and 8 female SP-A KO mice) were divided into 12 groups with 4 animals per group (Table 1 ).
Mice were exposed to either 2 parts/million (ppm) ozone or to filtered air (FA) for 3 h as described previously (13 (link), 53 (link)). In brief, four mice were put into a glass exposure vessel with stainless steel wire mesh lids and then placed in a closed glass exposure chamber. Exposures to FA were conducted in parallel at room temperature and 50% humidity using the ozone exposure system as described previously (50 (link), 53 (link), 54 (link)). After FA or ozone exposure, the mice were immediately infected with K. pneumoniae bacteria. This was done by anesthetizing the mice, surgically exposing the trachea, and instilling 50 μl of bacterial suspension. The mice were sacrificed 24 h after infection by anesthetizing them with an intramuscular injection of a 40 μl mixture of Ketamine/HCl a (Ketaset, Fort Dodge Animal Health, IO) and Xylazine (XYLA-JECT, Phoenix Pharmaceuticals Inc., St. Joseph, MO) and exsanguination. The lungs were lavaged with normal saline.
Mice were exposed to either 2 parts/million (ppm) ozone or to filtered air (FA) for 3 h as described previously (13 (link), 53 (link)). In brief, four mice were put into a glass exposure vessel with stainless steel wire mesh lids and then placed in a closed glass exposure chamber. Exposures to FA were conducted in parallel at room temperature and 50% humidity using the ozone exposure system as described previously (50 (link), 53 (link), 54 (link)). After FA or ozone exposure, the mice were immediately infected with K. pneumoniae bacteria. This was done by anesthetizing the mice, surgically exposing the trachea, and instilling 50 μl of bacterial suspension. The mice were sacrificed 24 h after infection by anesthetizing them with an intramuscular injection of a 40 μl mixture of Ketamine/HCl a (Ketaset, Fort Dodge Animal Health, IO) and Xylazine (XYLA-JECT, Phoenix Pharmaceuticals Inc., St. Joseph, MO) and exsanguination. The lungs were lavaged with normal saline.
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