Chimeric mice expressing MyD88 in all cells (WT⇒WT), restricted to myeloid cells (WT⇒<i>Myd88</i>^<i>-/-</i>), restricted to non-myeloid cells (<i>Myd88</i>^<i>-/-</i>⇒WT), or in no cells (<i>Myd88</i>^<i>-/-</i>⇒<i>Myd88</i>^<i>-/-</i>).

<p>Chimeric mice expressing MyD88 in all cells (WT⇒WT), restricted to myeloid cells (WT⇒<i>Myd88</i>^<i>-/-</i>), restricted to non-myeloid cells (<i>Myd88</i>^<i>-/-</i>⇒WT), or in no cells (<i>Myd88</i>^<i>-/-</i>⇒<i>Myd88</i>^<i>-/-</i>).</p

Cytokine concentrations from the right lung of mice subjected to 1 hr of left lung warm ischemia and 4 hr of reperfusion (n = 12/grp for C57BL/6 and <i>Tlr4</i>^<i>-/-</i>; n=10 for <i>Myd88</i>^<i>-/-</i>).

<p>Sham surgery mice (n = 4/grp) were not subjected to left lung ischemia. *p ≤ 0.05 when compared with the C57BL/6 mice. ^†p ≤ 0.05 when compared with <i>Tlr4</i>^<i>-/-</i> mice.</p

Myeloperoxidase activity in left lung (A) and right lung (B) homogenates of mice subjected to 1 hr of left lung warm ischemia and 4 hr of reperfusion (n = 12/grp for C57BL/6 and <i>Tlr4</i>^<i>-/-</i>; n=11 for <i>Myd88</i>^<i>-/-</i>).

<p>Sham surgery mice (n = 4/grp) were not subjected to left lung ischemia. *p ≤ 0.05 when compared with C57BL/6 mice.</p

IL-8 release by HEK293T cells expressing murine TLR4, CD14, and MD2.

<p>Cells were stimulated with serum, right lung BAL fluid, or left lung BAL fluid collected from mice subjected to 1 hr of left lung ischemia followed by 4 hr of reperfusion. Cells were treated with medium only for a negative control or with LPS (10 ng/ml) for a positive control. Each experiment was carried out with and without polymixin B (50 µg/ml).</p

Lung permeability as measured by total protein (A), 70kD Dextran (B), and IgM (C) in bronchoalveolar lavage fluid.

<p>(n = 6/grp, *p ≤ 0.05 compared with C57BL/6 and, ^†p ≤ 0.05 compared with <i>Tlr4</i>^<i>-/-</i>).</p

Peak airway pressures measured at various times during the experimental protocol for mice subjected to either 1 hr of left lung warm ischemia and 4 hr of reperfusion (-IR, n = 12/grp for C57BL/6 and <i>Tlr4</i>^<i>-/-</i>; n=10 for <i>Myd88</i>^<i>-/-</i>) or sham thoracotomy and 5 hr of mechanical ventilation (-Sham, n=4/grp).

<p>*p ≤ 0.05 between <i>MyD88</i>^<i>-/-</i> and <i>Tlr4</i>^<i>-/-</i> mice subjected to sham surgery at specific times.</p

Cytokine concentrations from the left lung of mice subjected to 1 hr of left lung warm ischemia and 4 hr of reperfusion (n = 12/grp for C57BL/6 and <i>Tlr4</i>^<i>-/-</i>; n=10 for <i>Myd88</i>^<i>-/-</i>).

<p>Sham surgery mice (n = 4/grp) were subjected to the same procedure except for left lung ischemia. *p ≤ 0.05 when compared with C57BL/6 mice. ^†p ≤ 0.05 when compared with <i>Tlr4</i>^<i>-/-</i> mice.</p

Neutrophil chemotaxis.

<p>Neutrophils were isolated from vitamin D-sufficient (VDS) or –deficient (VDD) mouse bone marrow for chemotaxis measurement and flow cytometry. (<b>A</b>) VDS neutrophil chemotaxis towards BAL fluid collected from VDS and VDD mice 1 day after LPS stimulation; Chemotaxis of neutrophils isolated from VDS and VDD mice towards varying concentrations of (<b>B</b>) KC/CXCL1 (N = 5 per group) and (<b>C</b>) complement C5a (N = 3 per group); (<b>D</b>) Percentage of CXCR2 positive cells in neutrophils isolated from bone marrow of VDS and VDD mice (N = 4 per group); (<b>E</b>) Expression of CXCR2 as determined by mean fluorescent intensity measured with flow cytometry of CXCR2+ cells from bone marrow of VDS and VDD mice (N = 4 per group).</p

Representative histology sections.

<p>400× magnfication of hematoxylin and eosin stained sections of right lung lobes collected from vitamin D sufficient (VDS) and deficient (VDD) mice after exposure to PBS or to LPS for varying lengths of time.</p

Lung inflammation 1 day after either LPS instillation.

<p>Vitamin D-sufficient (VDS) or –deficient (VDD) treated with either LPS or PBS. N = 7 to 9 per group. (<b>A</b>) Bronchoalveolar lavage (BAL) fluid total cell count, p<0.05 comparing VDS and VDD mice after LPS instillation; (<b>B</b>) BAL fluid neutrophil count, p = 0.09 comparing VDS and VDD mice after LPS; (<b>C</b>) BAL fluid KC/CXCL1 concentration; (<b>D</b>) BAL fluid MIP2/CXCL2 concentration.</p