Fig 1.
(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.
Fig 2.
Effect of adipose-derived stem cells (ASCs) on airway hyperresponsiveness (AHR) and inflammatory cells in the bronchoalveolar lavage fluid (BALF).
ASCs significantly decreased AHR and the number of total inflammatory cells and eosinophils in asthmatic mice. Treatment with the PGE2 inhibitor or TGF-β neutralizing Ab eliminated the reduction in AHR (A, C) and total cell and eosinophil counts (B, D) induced by ASC treatment. Data are expressed as the mean ± SEM of four independent experiments each performed in triplicate. *,†,§,¶,††,ǁǁ,***,††† p<0.001, ‡ p = 0.032, ǁ p = 0.045, ** p = 0.027, ‡‡ p = 0.032, §§ p = 0.005, ¶¶ p = 0.005.
Fig 3.
Effects of adipose-derived stem cells (ASCs) on lung inflammation and goblet cell hyperplasia.
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 p<0.001, † p = 0.020, ‡ p = 0.024, ¶ p = 0.003, †† p = 0.030, ‡‡ p = 0.035, ¶¶ p = 0.005.
Fig 4.
Effect of adipose-derived stem cells (ASCs) on serum levels of immunoglobulin.
Systemic administration of ASCs resulted in a significant decrease in total and OVA-specific IgE in asthmatic mice. (A) A PGE2 inhibitor significantly increased OVA-specific IgE and IgG1 in the OVA+ASC group. (B) TGF-β neutralizing Ab resulted in significant increases in total IgE and IgG1 and OVA-specific IgE and IgG1 in the OVA+ASC group. Data are expressed as the mean ± SEM of four independent experiments each performed in triplicate. *,‡,ǁ,††,ǁǁ,†††,ǁǁǁ,†††† p<0.001, †,*** p = 0.028, § p = 0.038, ¶,‡‡,¶¶ p = 0.008, **,§§ p = 0.009, ‡‡‡ p = 0.027, §§§ p = 0.037, ¶ p = 0.007, **** p = 0.035, ‡‡‡‡ p = 0.023, §§§§ p = 0.032.
Fig 5.
Effect of adipose-derived stem cells (ASCs) on cytokine levels in the bronchoalveolar lavage fluid.
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,¶,**, §§§,§§§§, ¶¶¶¶¶ p<0.001, †,****** p = 0.007, ‡ p = 0.027, ††,¶¶¶ p = 0.028, ‡‡ p = 0.030, §§ p = 0.010, ǁǁ p = 0.022, ¶¶ p = 0.032, ***,‡‡‡ p = 0.038, ††† p = 0.049, ǁǁǁ,†††† p = 0.003, ****,‡‡‡‡ p = 0.004, ǁǁǁǁǁ p = 0.026, ¶¶¶¶ p = 0.029, ***** p = 0.006, ††††† p = 0.012, ‡‡‡‡‡ p = 0.036, §§§§§ p = 0.042, ǁǁǁǁǁ p = 0.046.
Fig 6.
Effect of adipose-derived stem cells (ASCs) on cytokine levels in the lung draining lymph nodes.
ASCs treatment significantly decreased IL-4, IL-5, and IL-13 levels but increased IFN-γ, IL-10 and TGF—β levels in asthmatic mice. However, 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. *,*** p<0.001, †,†††† p = 0.003, ‡,§,ǁǁǁǁ p = 0.048, ǁ p = 0.037, ¶ p = 0.035, **,**** p = 0.004, ††,§§§ p = 0.009, ‡‡ p = 0.032, §§,¶¶ p = 0.038, ǁǁ p = 0.006, ‡‡‡ p = 0.007, ‡‡‡ p = 0.002, ǁǁǁ p = 0.040, ¶¶¶ p = 0.029, ‡‡‡ p = 0.049, §§§ p = 0.020, ¶¶¶¶ p = 0.008.
Fig 7.
Effects of adipose-derived stem cells (ASCs) on T cells in the lung draining lymph nodes.
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.