Comparison of DC surface marker activation and the production of Th2-related cytokine expressing cells.

Activated BMDCs surface marker (A) and IL-4 expressing T cells (B) were compared between the WT and TLR2 KO groups. (n = 3/group, 3 independent experiments, *; p p p < 0.001).</p

Comparison of the histopathological changes after KA/E2 treatment in WT and TLR2 KO mice.

Tissue inflammation was observed in the images of lung sections after H&E or PAS staining (A). Representative inflammation scoring and PAS-positive cells (mean ± SEM). A grade of 0 indicates no detectable inflammation and a grade 4 indicates high percentages of airways and blood vessels in section cuffing by inflammatory cells (0 = normal tissue; 1 = 75%). Severity scoring was based on the thickness of the bronchi or vessels surrounding inflammatory cells (0 = no cells; 1 = 1–3; 2 = 4–6; 3 = 7–9 cells thick; 4 = 10 or more cells thick). Means of the p-value were calculated for comparison to the control (n = 5 mice/group, 3 independent experiments, *; p < 0.05).</p

KA/E2 induced allergic airway inflammation via TLR2.

Lung inflammation was induced via intranasal inoculation of 4x105 KA/E2 trophozoites (A). Airway resistance values in response to methacholine (0 to 50 mg/ml) were compared between WT and TLR2 KO mice treated with KA/E2 (B). Differential inflammatory cells were counted in BAL from WT or TLR2 KO mice using a microscope (C). (n = 5 mice/group, 3 independent experiments, *; p < 0.05).</p

Comparison of the Th2 cytokine expression level after treatment with KA/E2 in WT and TLR2 KO mice.

Th2 related cytokine (IL-4, IL-5, and IL-13) concentrations in the BALF (A) and culture supernatants of CD3-stimulated lymphocytes from LLN (B) were determined using ELISA. The plates were read at 450 nm on a standard ELISA reader, VICTOR 3 (Each p-value was determined using t-test methods compared to the control) (n = 5 mice/group, 3 independent experiments, *p p < 0.01).</p

Evaluation of the inflammatory-related gene expression owing to KA/E2 ES-P in primary lung cells from WT and TLR2 KO mice.

Assessment of the expression of inflammatory response related genes in primary lung cells from WT and TLR2 KO mice. (n = 3/group, 3 independent experiments, *; p p p < 0.001).</p

KA/E2 ES-P induced inflammatory-related gene expression by TLR2 in MLE12 cells.

Comparison of the expression of inflammatory response-related genes in MLE 12 cells from the KA/E2 ES-P group or TLRs antagonist pre-treated group. (n = 3/group, 3 independent experiments, *; p p p < 0.001).</p

The experimental protocol.

<p>(A) Mice were sensitized on days 0, 1, 7, and 8 by intraperitoneal injection of ovalbumin (OVA) and challenged intranasally on days 14, 15, 21, and 22. Purified adipose-derived stem cells (ASCs; 1 × 10⁶) were injected via the tail vein on days 12, 13, 19, and 20. PGE2 and TGF-β were blocked by intraperitoneal injection of a PGE2 inhibitor or anti-TGF-β-Ab on days 13, 14, 15, 16, 17, 20, 21, 22, and 23. (B) The mice were divided into five treatment groups.</p

Effect of adipose-derived stem cells (ASCs) on cytokine levels in the bronchoalveolar lavage fluid.

<p>IL-4, IL-5, and IL-13 were significantly higher in the OVA group than PBS group. ASC treatment significantly decreased IL-4, IL-5, and IL-13 but increased IL-10 and TGF—β in asthmatic mice. However, the PGE2 inhibitor (A) or TGF-β neutralizing Ab (B) eliminated these immunomodulatory effects of ASCs. Data are expressed as the mean ± SEM of four independent experiments each performed in triplicate. *,§,ǁ,¶,_**, §§§,§§§§, ¶¶¶¶¶ <i>p</i><0.001, †,_******<i>p</i> = 0.007, ‡ <i>p</i> = 0.027, ††,¶¶¶ <i>p</i> = 0.028, ‡‡ <i>p =</i> 0.030, §§ <i>p =</i> 0.010, ǁǁ <i>p</i> = 0.022, ¶¶ <i>p</i> = 0.032, ***,‡‡‡ <i>p</i> = 0.038, ††† <i>p =</i> 0.049, ǁǁǁ,†††† <i>p</i> = 0.003, ****,‡‡‡‡ <i>p =</i> 0.004, ǁǁǁǁǁ <i>p =</i> 0.026, ¶¶¶¶ <i>p</i> = 0.029, ***** <i>p</i> = 0.006, ††††† <i>p</i> = 0.012, ‡‡‡‡‡ <i>p</i> = 0.036, §§§§§ <i>p =</i> 0.042, ǁǁǁǁǁ <i>p =</i> 0.046.</p

Effects of adipose-derived stem cells (ASCs) on lung inflammation and goblet cell hyperplasia.

<p>ASCs treatment decreased the infiltration of eosinophils and PAS-positive cells around the airway and blood vessel in asthmatic mice (H&E, PAS ×200). Blocking PGE2 (A) or TGF-β (B) eliminated the beneficial effect of ASCs on lung inflammation and goblet cell hyperplasia. Data are expressed as the mean ± SEM of four independent experiments each performed in triplicate. *,§,ǁ,**,§§,ǁǁ <i>p</i><0.001, † <i>p =</i> 0.020, ‡ <i>p</i> = 0.024, ¶ <i>p</i> = 0.003, †† <i>p =</i> 0.030, ‡‡ <i>p</i> = 0.035, ¶¶ <i>p</i> = 0.005.</p

Effects of adipose-derived stem cells (ASCs) on T cells in the lung draining lymph nodes.

<p>The CD4⁺ T cells were initially gated and the percentage of IFN-γ⁺, IL-4⁺, IL-10⁺, and CD25⁺ Foxp3⁺ T cells subsequently analyzed. When treating asthmatic mice with PGE2 inhibitor (A) or TGF-β neutralizing Abs (B), blocking of PGE2 and TGF-β prevented the increases in Foxp3⁺CD25⁺, IL-10⁺, and IFN-γ⁺ T cell populations and the decrease in the IL-4⁺ T cell population in the OVA+ASC group.</p