Increased lavage total protein level after ozone exposure was independent of TLR4, MyD88, and TIRAP (*p<0.01, vs. FA exposed group; N = 5–6 per group).

<p>Increased lavage total protein level after ozone exposure was independent of TLR4, MyD88, and TIRAP (*p<0.01, vs. FA exposed group; N = 5–6 per group).</p

Ozone inhalation increased the level of pro-inflammatory factors in alveolar lavage fluid in a manner partially dependent on TLR4, MyD88, and TIRAP.

<p>The level of (<b>A</b>) KC, (<b>B</b>) IL-1β, (<b>C</b>) IL-6, (<b>D</b>) MCP-1 and (<b>E</b>) TNFα in BALF from air and ozone-exposed WT, TLR4−/−, MyD88−/− and TIRAP−/− mice were measured by luminex beads (*p<0.05, vs. FA exposed group; #p<0.05, vs. O₃-exposed group comparisons between strains; N = 4–5 per group).</p

Ozone inhalation increased airway sensitivity to methacholine challenge and was dependent on the TLR4-MyD88-TIRAP signaling pathway.

<p>Animals were exposed to filtered air (FA) or 1 ppm of ozone for 3 hours. Airway responsiveness to methacholine challenge was measured 24 h later. <b>A</b>) Ozone-induced AHR was increased in WT mice but not in TLR4−/− mice, <b>B</b>) MyD88−/−mice, or <b>C</b>) TIRAP−/− mice (*p<0.05 vs. FA-WT; #p<0.05 vs. O3-WT, N = 5–6 per group).</p

HA challenge was not sufficient for either neutrophilic inflammation or epithelial injury.

<p>(<b>A</b>) There were no observed differences in cellular inflammation in the airspace 2 hours after direct challenge to HA, (<b>B</b>) When compared to ozone challenge, HA challenge had no observed effect on the level of BALF total protein (*p<0.05, vs. FA exposed group, N = 5).</p

Level of HA in the BALF was increased after exposure to ozone.

<p>HA levels were significantly increased in all ozone-exposed groups (*p<0.01, vs. FA exposed group; #p<0.05, vs. O3-exposed WT, group, N = 4–5 per group).</p

Ozone exposure increased neutrophilic lung inflammation in a manner partially dependent on MyD88, but not TLR4 and TIRAP.

<p>Ozone-exposed mice demonstrated increased neutrophil cell counts in BALF when compared to air-exposed animals. Neutrophil recruitment to the airspace was independent of TLR4 (<b>A</b>) and TIRAP (<b>C</b>), but was partially dependent of MyD88 (<b>B</b>) (* p<0.05, vs. FA exposed group; # p<0.05 vs. O₃-WT, N = 4–6 per group).</p

Direct challenge to hyaluronan fragments increased the level of pro-inflammatory factors in alveolar lavage fluid in a manner partially dependent on TLR4, MyD88, and TIRAP.

<p>The level of (<b>A</b>) KC, (<b>B</b>) IL-1β, (<b>C</b>) IL-6, (<b>D</b>) MCP-1 and (<b>E</b>) TNFα in BALF from vehicle or HA-challenged WT, TLR4−/−, MyD88−/− and TIRAP−/− mice were measured by luminex beads (*p<0.05, vs. FA exposed group; #: p<0.05, vs. O₃-exposed group comparisons between strains; N = 4 per group).</p

Airway reactivity to HA challenge was dependant on TLR4, MyD88, and TIRAP.

<p>HA increased AHR in WT mice, but not in (<b>A</b>) TLR4−/−mice, (<b>B</b>) MyD88−/−mice, or (<b>C</b>) TIRAP−/− mice (*p<0.05 vs. vehicle-WT; #p<0.05 vs. HA-WT, N = 5 per group).</p

Eosinophil mediators are increased in the absence of SP-A.

<p>On day 28 of the Ova+Mp model, lungs were harvested and <b>A</b>) IL-5 and <b>B</b>) EAR were assessed by RT-PCR. <b>C</b>) Histochemical staining for EPO positive eosinophils was done 1 day after Mp infection and is representative of 3 experiments, n = 5/group. <b>D</b>) Total EPO activity in BAL and lung tissue from Ova+Mp mice was determined via colorimetric assay and absorbance read at 492 nm. n = combined 3 experiments, *p<.05, **p<.01.</p

Recruitment of inflammatory cells is enhanced in Mp-infected SP-A^{<b>−/−</b>} allergic mice.

<p><b>A–F</b>) Cells in the BAL were examined by cell surface labeling, as described in the methods section, and flow cytometry 3 days after Mp infection (day 28 of the model). Macs (macrophages), Ex-Macs (exudative macrophages), IMs (inflammatory monocytes), DCs (dendritic cells), PMNs (neutrophils), Eos (eosinophils). n = at least 12 mice/group, 3 experiments combined, *p<.05, **p<.01.</p