Airway epithelial cell-induced changes in DC expression of Fcγ receptor genes.

<p>After 5 days of culture in the presence or absence of AEC, DC were sorted by flow cytometry. RNA from 15 independent experiments was extracted, and expression of Fc gamma receptor genes was determined using quantitative real-time PCR. **p<0.01; ***p<0.001.</p

Airway epithelial cell-induced changes in DC expression of selected immune response genes.

<p>(A). After 5 days of culture in the presence or absence of AEC, DC were sorted by flow cytometry. RNA from 15 independent experiments was extracted, and expression of immune response genes was determined using quantitative real-time PCR. **p<0.01; ***p<0.001. (B) Cell surface expression of B7-H1 and ICAM-1 was determined by flow cytometry. Cells staining with specific antibody and isotype control antibodies are shown. Histograms from a representative experiment are shown. Similar changes were seen in all 8 experiments performed.</p

DC and airway epithelial cell expression of CD200R1 and CD200.

<p>After 5 days of culture in the presence or absence of AEC, cell surface expression of CD200R1 on DC and CD200 on AEC was determined by flow cytometry. Histograms from a representative experiment are shown. Similar changes were seen in all 6 experiments performed.</p

Expression of type I interferon and IL-6 in AEC co-cultured with MDDC.

<p>AEC were cultured alone, in the presence or absence of GM-CSF + IL-4, or with MDDC. Following 5 days of culture RNA was extracted from sorted AEC and relative expression IFNα2 and IFNβ. *indicates p<0.05 relative to AEC alone (N = 6). IFNα protein was undetectable in culture supernatants. IL-6 protein data are from two experiments.</p

Long-term changes in total cell numbers of, and co-stimulatory molecule expression by, of respiratory APC populations following IAV infection in early life.

<p>Juvenile (28 day old) BALB/c mice were infected i.n. with a weight-adjusted dose of IAV as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111520#s4" target="_blank">Materials and Methods</a>, then respiratory tissues harvested 35 days later as 8 week-old adults. <i>(A)</i> Representative FACS profiles showing gating for AMDC in tracheal tissue of control (left) and IAV infected mice (right). <i>(B and C)</i> Total AMDC numbers <i>(B)</i> and percentage changes in co-stimulatory marker expression <i>(C)</i>, expressed a percentage change from control mice. <i>(D)</i> Representative FACS profiles showing gating for PLDC in parenchymal lung tissue of control (left) and IAV infected mice (right). <i>(E and F)</i> Total PLDC numbers <i>(E)</i> and percentage changes in co-stimulatory marker expression <i>(F)</i>, expressed a percentage change from control mice. <i>(G)</i> Representative FACS profiles showing gating for PLMac in parenchymal lung tissue of control (left) and IAV infected mice (right). <i>(H and I)</i> Total PLMac numbers <i>(H)</i> and percentage changes in co-stimulatory marker expression <i>(I)</i>, expressed a percentage change from control mice. Data are shown for 4 independent infection experiments, using pools of tissue from 4 to 5 mice for each experiment. *  =  <i>p</i><0.05; **  =  <i>p</i><0.01.</p

Alum-induced chemokine production with increasing age in infancy.

<p>Whole blood samples from PNG infants aged 1–3 months (n = 10, <i>white bars</i>), 4–6 months (n = 9, <i>light grey bars</i>), 7–12 months (n = 10, <i>dark grey bars</i>) or 13–18 months (n = 9, <i>black bars</i>) were stimulated with Alum. Presented are the geometric means and 95% confidence intervals (pg/mL) for each age group for background-adjusted chemokine responses. Significance level is indicated where p<0.10.</p

Infant characteristics.

<p>Infant characteristics.</p

Time course of changes in the expression of CD11b on respiratory DC subsets in anatomical compartments of the respiratory tract following IAV infection.

<p><i>(A and B)</i> Percentage frequency of CD11b^lo AMDC <i>(A)</i> and CD11b^hi AMDC <i>(B)</i> amongst total AMDC (gated as per Fig. 4A) in IAV infected (closed circles) and control mice (open circles). <i>(C and D)</i> Percentage frequency of CD11b^lo PLDC <i>(A)</i> and CD11b^hi PLDC <i>(B)</i> amongst total PLDC (gated as per Fig. 4D) in IAV infected (closed circles) and control mice (open circles). Data are means +/− SEM for 3 independent infection experiments using pools of tissue from 3 to 4 mice for each experiment. *  =  <i>p</i><0.05; **  =  <i>p</i><0.01; ***  =  <i>p</i><0.001.</p

Innate TNF-α responses in relation to increasing age in infancy.

<p>Presented are the geometric means and 95% confidence intervals (pg/mL) for background-adjusted TNF-α responses. Mann-Whitney U tests for significant differences in log-transformed TNF-α levels compared to the “1–3 months” age group; and Spearman rho tests for significant correlations between log-transformed TNF-α levels and ordered age groups were conducted. Significance level is indicated where p<0.05 only (<b>*</b> p = 0.049).</p>♦<p>denotes LPS co-stimulation used.</p

Innate IL-10 responses in relation to increasing age in infancy.

<p>Presented are the geometric means and 95% confidence intervals (pg/mL) for background-adjusted IL-10 responses. Mann-Whitney U tests for significant differences in log-transformed IL-10 levels compared to the “1–3 months” age group; and Spearman rho tests for significant correlations between log-transformed IL-10 levels and ordered age groups were conducted. Significance level is indicated where p<0.05 only (<b>*</b> p = 0.046; <b>‡</b> p = 0.022).</p>♦<p>denotes LPS co-stimulation used.</p