Percentage of cells that express VEGFR2 after CpG culture.

<p>Epithelial VEGFR2 expression after 4 hours (<b>A</b>) and 24 hours (<b>B</b>) of culture with vehicle/CpG (0.1 µM; 0.3 µM and 1.0 µM). Expression on turbinate epithelial cells from healthy controls, turbinate and polyp epithelial cells from patients with CRSwNP, analysed using flow cytometry. Results are presented as mean ± SEM, <i>n</i> = 5, *<i>P</i><0.05, **<i>P</i><0.01 (unstimulated vs. CpG stimulated), ##<i>P</i><0.01 (unstimulated turbinate tissue from patients vs. unstimulated turbinate tissue from healthy controls and polyp tissue from patients).</p

Epithelial TLR9 expression after CpG stimulation.

<p>Epithelial TLR9 expression after 4 hours (<b>A</b>) and 24 hours (<b>B</b>) of culture with vehicle/CpG (0.1 µM; 0.3 µM and 1.0 µM). Expression on turbinate epithelial cells from healthy controls, turbinate and polyp epithelial cells from patients with CRSwNP, analysed using flow cytometry. Results are presented as mean ± SEM, <i>n</i> = 6, **<i>P</i><0.01 (unstimulated vs. CpG stimulated), ##<i>P</i><0.01 (unstimulated turbinate tissue from patients vs. unstimulated turbinate tissue from healthy controls and polyp tissue from patients).</p

Cytokine secretion from the nasal mucosa after CpG stimulation.

<p>CpG induced/reduced cytokine release in culture with turbinate tissue from healthy controls and turbinate tissue from patients with CRSwNP. Samples analysed after culture of 4 hours (<i>n</i> = 4) or 24 hours (<i>n</i> = 5–8). Levels of GM-CSF (<b>A</b>), IL-6 (<b>B</b>), G-CSF (<b>C</b>) and MIP-1β (<b>D</b>) were analysed with Luminex. Data presented as mean ± SEM, *<i>P</i><0.05; **<i>P</i><0.01 (unstimulated vs. CpG stimulated), #<i>P</i><0.05 (unstimulated turbinate tissue from healthy controls vs. unstimulated turbinate tissue from patients).</p

Epithelial TLR9 expression in turbinate tissue from healthy controls as well as turbinate tissue and polyp tissue from patients with CRSwNP.

<p>Expression of TLR9 on turbinate epithelial cells from healthy controls compared to turbinate and polyp epithelial cells from patients with CRSwNP, <i>n</i> = 5 (<b>A</b>). Intracellular staining for TLR9 (open histogram, black line) and isotype control (filled histogram) on turbinate epithelial cells from a healthy control (<b>B</b>), turbinate epithelial cells (<b>C</b>) and polyp epithelial cells from a patient (<b>D</b>), analysed using flow cytometry. Results are presented as mean ± SEM, **<i>P</i><0.01.</p

TLR9 and VEGFR2 expression after nasal challenge with CpG.

<p>Eight patients with CRSwNP were nasally challenged with either CpG (<i>n</i> = 4) or placebo (<i>n</i> = 4). Biopsies from the turbinate tissue were obtained during FESS, 24 hours later. The nasal turbinate epithelial cells were stained intracellularly for expression of TLR9 (<b>A</b>) and VEGFR2 (<b>B</b>) and analysed using flow cytometry. Results are presented as mean ± SEM, *<i>P</i><0.05.</p

Poly(I:C) and LPS induced cytokine release in BALF.

<p>Levels of (A) IL-5, (B) IL-12, (C) MCP-1, (D) KC, (E) MIG and (F) VEGF in BALB/c mice treated during 4 days with vehicle (white), poly(I:C) (black) or LPS (gray) i.n. Results are expressed as mean ± SEM. PBS n = 10; poly I:C n = 12; LPS n = 13; ***p<0.001.</p

LPS and poly(I:C) induced airway hyperresponsiveness.

<p>(A) Changes in lung resistance (R_L) and (B) compliance (C_L) in response to methacholine in BALB/c mice treated during 4 days with vehicle (PBS), poly(I:C) or LPS i.n.. Results are expressed as mean ± SEM. *p<0.05, ***p<0.001 vs. PBS.</p

Poly(I:C) and LPS induced cell recruitment in lung tissue.

<p>Haematoxylin and eosin stained cells in BALB/c mice treated i.n. during 4 days with (A) vehicle, (B) poly(I:C) (mainly neutrophils) and (C) LPS (mainly lymphocytes). Magnification is ×200. (D) Semi quantitative grading score of cell infiltration in the lung.</p

The nasal epithelium expresses TLR3, TLR7, TLR9, RIG-I and MDA-5.

<p>Sections of nasal biopsies were incubated with antibodies against TLR3 (<b>A</b>), TLR7 (<b>B</b>), TLR9 (<b>C</b>), RIG-I (<b>D</b>), and MDA-5 (<b>E</b>) and visualized by 3, 3′-diaminobenzidine (brown). In control slides (<b>F</b>), N-series universal negative control reagent was used. All sections were accompanied with a square magnification. All slides were counterstained with haematoxylin (blue). The figure shows one representative biopsy out of four (3 male, 1 female). The arrows indicate positive stained cells.</p

Poly(I:C) and LPS induced cell recruitment in BALF.

<p>(A, C) Absolute and (B, D) relative cell numbers in BALF in BALB/c mice (A, B) respectively MyD88deficient C57BL/6 mice (C, D) treated during 4 days with vehicle (PBS), poly(I:C) or LPS i.n.. Results are expressed as mean ± SEM. *p<0.05.</p