CFU recovered in BAL cell culture supernatant following OPKA.

†<p>Adjusted for clustering within participants.</p

rhIL-17A stimulated alveolar macrophages show increased killing of opsonised pneumococci.

<p>Human alveolar macrophages from non-colonised volunteers were exposed to opsonised D39 pneumococci in the presence of rhIL-17A or a vehicle control (v/v). Percentage changes in CFU following rhIL-17A (<i>y</i>-axis) treatment are shown as increase or decrease relative to vehicle, which was set at 0% for comparison (<i>x</i>-axis). * = <i>p</i><0.05, ** = <i>p</i><0.01 vs 12.5 ng/ml of rhIL-17A.</p

Cellular profile of BAL collected from the pneumococcal non-colonised and colonised groups.

<p>Cellular profile of BAL collected from the pneumococcal non-colonised and colonised groups.</p

BAL cells stimulated with pneumococci <i>ex vivo</i> elicit large quantities of IL-17A.

<p>BAL cells from non-colonised (open bars, n = 6) or colonised (closed bars, n = 7, 1x 23F, 6x6B colonised) volunteers were stimulated with pneumococci (6B) or left untreated (NS) for 20 hours. Following stimulation <b>A</b> IL-17A and <b>B</b> TNF were measured in pg/ml (mean±SD) in cell culture supernatant by ELISA. * = <i>p</i><0.05.</p

Pneumococcal carriage increases the percentage of IL-17A⁺/TNF⁺ CD4⁺ memory T-cells in BAL.

<p>Sub-population analysis of 6B stimulated BAL CD4⁺ memory T-cell responses from <b>A</b> non-colonised (n = 9) and <b>B</b> colonised volunteers (n = 11). The percentage of triple, double or single CD4⁺ memory T-cells producing either TNF, IL-17A and/or IFNγ (<i>x</i>-axis) are shown. Pneumococcal-responding double-producing (IL-17A⁺/TNF⁺) CD4⁺ memory T-cells constitute the dominant phenotype following pneumococcal colonisation. Responses shown are background subtracted. * = <i>p</i><0.05; ** = <i>p</i><0.01 vs non-colonised in <b>A</b>.</p

Study volunteer and colonisation details.

<p> <b>N.D. – Not determined.</b></p><p> <b>N/A – Not applicable.</b></p>†<p> <b>- Values given are background (media control) subtracted from cells stimulated with HK-6B.</b></p

Pneumococcal carriage increases the percentage of pneumococcal-responding IL-17A⁺ CD4⁺ memory T-cells in BAL and blood.

<p><b>A</b> BAL CD4⁺ memory T-cell and <b>B</b> Blood CD4⁺ memory T-cell responses from volunteers without (−, BAL n = 9, blood n = 8) or with (+, BAL n = 11, blood n = 10) experimental pneumococcal carriage (indicated on <i>x</i>-axis). Cells were stimulated <i>ex vivo</i> with or without (not shown) influenza or pneumococcal proteins as indicated. TNF⁺, IL-17A⁺ or IFNγ⁺ responses shown are background subtracted and measured as % of total CD4⁺ memory T-cells. Note the change in scale for the <i>y</i>-axis for each cytokine ns = not significant, * = <i>p</i><0.05; ** = <i>p</i><0.01 vs non-colonised volunteers.</p

Primers and probes used for qPCR assays.

a<p>All probes were labeled with Hex at 5′-end and Black Hole Quencher (BHQ) at the 3′-end with the exception for hpdPr762, which was labeled with BHQ at an internal “T” and SpC6 at the 3′-end.</p

Percent carrying Hi (A), Sa (B), and Mc (C) at 4 time points and density of Sa colonization at baseline and 14 days (D) segregated by success of pneumococcal colonization.

<p>Numbers of subjects carrying each bacteria ranged from 11<b>–</b>17 for <i>H. influenzae</i>, 5<b>–</b>10 for <i>M. catarrhalis</i>, and 13<b>–</b>14 for <i>S. aureus</i>. Asterisk indicates <i>p</i> = 0.008.</p

Density of Sp (A), Hi (B), Sa (C), and Mc (D) in nasal wash samples at four time points.

<p>Boxplots of log bacterial density (in genomes/ml) of all samples with >100 genomes/ml at each time point. Boxplots indicate median, interquartile range, and range with circles indicating outlier values. Asterisk indicates <i>p</i> = 0.003.</p