Level of Foxp3 expression.

<p>The specific mRNAs in OVA-stimulated mice splenocytes (incubated with OVA for 72hrs) were quantified by qRT-PCR. The results are presented as mean mRNA expression (mean±SE, n = 8 for each). The relative levels of Foxp3 expression were calculated by referring to the HPRT (hypoxanthine guanine phosphoribosyltransferase) in each sample. AD without ASIT: OVA-sensitized untreated mice; ASIT with OVA: OVA-sensitized mice treated by ASIT with OVA; ASIT with sOVA: OVA-sensitized mice treated by ASIT with sOVA; placebo: PBS-sensitized mice. *, p<0.05.</p

Levels of cytokines.

<p>The levels of cytokines in supernatants of OVA-stimulated mice splenocytes (incubated with OVA for 72hrs) were quantified by ELISA. The results are presented as mean IL concentration (mean±SE, n = 8 for each). AD without ASIT: OVA-sensitized untreated mice; ASIT with OVA: OVA-sensitized mice treated by ASIT with OVA; ASIT with sOVA: OVA-sensitized mice treated by ASIT with sOVA; placebo: PBS-sensitized mice. Level of (a) the IL-4, (b) IL-5, (c) IL-12, (d) IFN-γ, and (e) IL-17; (f) IL-4/IFN-γ ratio in mice with ASIT. *, p<0.05 versus “AD without ASIT”; **, p<0.05 versus “placebo”; #, p<0.05 versus “ASIT with OVA”.</p

Analysis of OVA-specific IgG.

<p>The levels of antibodies in sera obtained before, during, and after ASIT were detected and quantified by ELISA. The results are presented as mean antibodies concentrations (mean±SE, n = 8 for each). AD without ASIT: OVA-sensitized untreated mice; ASIT with OVA: OVA-sensitized mice treated by ASIT with OVA; ASIT with sOVA: OVA-sensitized mice treated by ASIT with sOVA; placebo: PBS-sensitized mice. (a) anti-OVA IgG1 response in mice with ASIT, (b) anti-OVA IgG2a response in mice with ASIT, (c) anti-OVA IgG response in mice with ASIT, (d) ratio of OVA-specific IgG1/IgG2a antibody in mice with ASIT, (*, p<0.05)</p

Histologic features of OVA- and sOVA-treated skin sites in BALB/c mice.

<p>(a) Mice sensitized with OVA and treated with PBS (“AD without ASIT”), (b) mice sensitized with OVA and treated with OVA (“ASIT with OVA”), (c) mice sensitized with OVA and treated with sOVA (“ASIT with sOVA”), and (d) mice sensitized with PBS (“placebo”). Skin sections were stained with H&E and examined at 100x. Scale bars 100 μm. There is marked hyperplasia of the epidermis, a dermal infiltrate (a-c). The cellular infiltrate consists of neutrophils, eosinophils, and lymphocytes. (e) Summary index of the main assessment criteria for histologic skin lesions.</p

Protocol of sensitization and ASIT with OVA or sOVA.

<p>Mice were sensitized with OVA (100 μg) or saline applied at 100 μl to a sterile patch. The patch was applied for 1-wk and then removed. ASIT was performed between the 1^st and 2^nd OVA applications by SC injection of increasing doses of OVA (a), sOVA (b), or PBS (as control). After ASIT, the mice had two 1-wk exposures to a patch separated by 2-wk intervals. The control group received PBS at the same time.</p

Change in levels of fractalkine in PBMCs following <i>in vitro</i> infection with RV16 and RV1B.

<p>Soluble fractalkine protein was measured in cell supernatants from PBMCs obtained from (A) non-asthmatic (n = 15) and (B) asthmatic (n = 15) subjects and compared between subject groups at 8hrs post infection (C). Fractalkine mRNA expression was measured in PBMC cell lysate cDNA obtained from (D) non-asthmatic and (E) asthmatic subjects. The results are expressed as mean ± SEM. Protein data were analysed by one-way ANOVA with Bonferroni post-test and mRNA by Kruskal Wallis with Dunn’s post test (*<i>P</i><0.05, ***<i>P</i><0.001).</p

Change in levels of fractalkine in BAL cells following <i>in vitro</i> infection with RV16 and RV1B.

<p>Soluble fractalkine protein was measured in cell supernatants from BAL cells obtained from (A) non-asthmatic (n = 15) and (B) asthmatic (n = 15) subjects and compared between subject groups at 8hrs post infection (C). Fractalkine mRNA expression was measured in BAL cell lysate cDNA obtained from non-asthmatic (D) and asthmatic (E) subjects. The results are expressed as mean ± SEM. Protein data were analysed by one-way ANOVA with Bonferroni post-test and mRNA by Kruskal Wallis with Dunn’s post test (**<i>P</i><0.01).</p

Change in soluble fractalkine in BAL fluid during experimental <i>in vivo</i> RV16 infection, related with upper respiratory symptom scores.

<p>Soluble fractalkine protein was measured in filtered BAL fluid collected at baseline and day 4 post RV16 infection from non-asthmatic (n = 10), mild-asthmatic (n = 11) and moderate-asthmatic (n = 14) subjects. (A) Data is presented as soluble fractalkine (pg/mL) per subject and horizontal bars for median levels for each group in BAL fluid obtained at baseline and day 4. Data were analysed within groups by Wilcoxon-matched pairs signed rank tests and between groups by Mann Whitney U test, *P<0.05. (B) Levels of fractalkine in BAL fluid on Day 4 were correlated with peak upper respiratory symptom scores for each subject infected using Pearson’s correlation (r = 0.289, <i>P</i> = 0.098).</p

14C11 inhibits major group HRV replication <i>in vitro</i>.

<p>Ohio HeLa cells were pre-incubated with serial dilutions of 14C11 or isotype control and infected with (A) HRV16, (B) HRV14 and (C) minor group HRV25. The antiviral effect of 14C11 was determined by CPE reduction assay and expressed as % of control. (D) IC₅₀s for 14C11 were determined for major HRVs by CPE assay as indicated in. Experiment (A), (B) and (C) were performed in sextuplicates and (D) is a pool of two independent experiments. Data are expressed as mean (± SEM).</p

14C11 does not inhibit inflammation induced via mechanisms independent of human ICAM-1.

<p>Mice were dosed intravenously with 14C11 24 hours prior to intranasal dosing with HRV1B, UV-inactivated HRV1B (UV) or HRV16 (n = 4 for tg− UV, tg− 1B, tg+ 16 iso and tg+ 16 14C11 groups; n = 6 for tg+ UV, tg+ 1B, tg+ 1B iso and tg+ 1B 14C11 groups). (A)Total BAL cells, neutrophils (on day 1) and lymphocytes (on day 4) were assessed with differentially stained cytospins. Data expressed as mean (± SEM). Significance was assessed by One-way ANOVA test with Bonferroni's Multiple Comparison test as post-test. ***p<0.001 vs UV-RV1B in transgenic negative mice; ^###p<0.001 vs UV-RV1B in transgenic positive mice; ^§p<0.05 and ^§§§p<0.001 vs isotype treated HRV16 infected transgenic positive mice. Data are representative of 2 independent experiments. (B) Groups of 7 mice were dosed intravenously with 14C11 24 hours prior to intranasal dosing with 1 µg LPS/mouse. Total BAL cells, lymphocytes and neutrophils were assessed with differentially stained cytospins day 1 after infection. Data are expressed as mean (± SEM). Significance was assessed by One-way ANOVA test with Bonferroni's Multiple Comparison test as post-test. *p<0.05, **p<0.01 and ***p<0.001 vs transgenic positive mice without treatment. Data are representative of 2 independent experiments.</p