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Small interfering RNA against CD86 during allergen challenge blocks experimental allergic asthma.

Asai-Tajiri Y, Matsumoto K, Fukuyama S, Kan-O K, Nakano T, Tonai K, Ohno T, Azuma M, Inoue H, Nakanishi Y - Respir. Res. (2014)

Bottom Line: However, it remains uncertain whether this pathway also has an essential role in the effector phase.These results suggest that local administration of CD86 siRNA during the effector phase ameliorates lines of asthma phenotypes.Targeting airway dendritic cells with siRNA suppresses airway inflammation and hyperresponsiveness in an experimental model of allergic asthma.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan. yukari-a@kokyu.med.kyushu-u.ac.jp.

ABSTRACT

Background: CD86-CD28 interaction has been suggested as the principal costimulatory pathway for the activation and differentiation of naïve T cells in allergic inflammation. However, it remains uncertain whether this pathway also has an essential role in the effector phase. We sought to determine the contribution of CD86 on dendritic cells in the reactivation of allergen-specific Th2 cells.

Methods: We investigated the effects of the downregulation of CD86 by short interfering RNAs (siRNAs) on Th2 cytokine production in the effector phase in vitro and on asthma phenotypes in ovalbumin (OVA)-sensitized and -challenged mice.

Results: Treatment of bone marrow-derived dendritic cells (BMDCs) with CD86 siRNA attenuated LPS-induced upregulation of CD86. CD86 siRNA treatment impaired BMDCs' ability to activate OVA-specific Th2 cells. Intratracheal administration of CD86 siRNA during OVA challenge downregulated CD86 expression in the airway mucosa. CD86 siRNA treatment ameliorated OVA-induced airway eosinophilia, airway hyperresponsiveness, and the elevations of OVA-specific IgE in the sera and IL-5, IL-13, and CCL17 in the bronchoalveolar lavage fluid, but not the goblet cell hyperplasia.

Conclusion: These results suggest that local administration of CD86 siRNA during the effector phase ameliorates lines of asthma phenotypes. Targeting airway dendritic cells with siRNA suppresses airway inflammation and hyperresponsiveness in an experimental model of allergic asthma.

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Related in: MedlinePlus

Distribution of Red-labeled siRNA in the trachea after allergen exposure. Texas Red-labeled siRNA was administrated intratracheally to sensitized mice. The mice were challenged with OVA 1 hour later. One hour (A) or 12 hours (B) after allergen exposure, lungs are extracted and stained with DAPI and FITC-conjugated anti-CD11c mAb. Red-labeled siRNA is located in green CD11c-positive cells (arrow).
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Fig3: Distribution of Red-labeled siRNA in the trachea after allergen exposure. Texas Red-labeled siRNA was administrated intratracheally to sensitized mice. The mice were challenged with OVA 1 hour later. One hour (A) or 12 hours (B) after allergen exposure, lungs are extracted and stained with DAPI and FITC-conjugated anti-CD11c mAb. Red-labeled siRNA is located in green CD11c-positive cells (arrow).

Mentions: In an attempt to assess how intratracheally administrated siRNA was distributed to the lung, anesthetized mice were administered with Texas Red-labeled non-coding siRNA and then challenged with OVA. One hour and 12 hours after the challenge, frozen sections were made and stained with FITC-labeled anti-CD11c mAb to detect DCs [15]. One hour after the OVA challenge, Red-labeled siRNA was detected on the airway epithelial layer in a band-like deposition pattern. Green-labeled DCs were clustered around the deposited siRNA (Figure 3A). The immunofluorescence of CD11c+cells in proximity of the red siRNA signal was observed in several sites per one section, mainly on the airway epithelial layer. Due to its characteristic distribution, this was unlikely a sectioning/deparaffinizing/staining artifact. Twelve hours after the challenge, the red dots of siRNA were distributed beneath the airway basement membrane and submucosal DCs, indicating that siRNA successfully went through the epithelial barrier (Figure 3B,C). Yellow fluorescence was observed in several sites, suggestive of siRNA taken up by DCs, although a mere topological stratification of fluorescence might not be excluded.Figure 3


Small interfering RNA against CD86 during allergen challenge blocks experimental allergic asthma.

Asai-Tajiri Y, Matsumoto K, Fukuyama S, Kan-O K, Nakano T, Tonai K, Ohno T, Azuma M, Inoue H, Nakanishi Y - Respir. Res. (2014)

Distribution of Red-labeled siRNA in the trachea after allergen exposure. Texas Red-labeled siRNA was administrated intratracheally to sensitized mice. The mice were challenged with OVA 1 hour later. One hour (A) or 12 hours (B) after allergen exposure, lungs are extracted and stained with DAPI and FITC-conjugated anti-CD11c mAb. Red-labeled siRNA is located in green CD11c-positive cells (arrow).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4216659&req=5

Fig3: Distribution of Red-labeled siRNA in the trachea after allergen exposure. Texas Red-labeled siRNA was administrated intratracheally to sensitized mice. The mice were challenged with OVA 1 hour later. One hour (A) or 12 hours (B) after allergen exposure, lungs are extracted and stained with DAPI and FITC-conjugated anti-CD11c mAb. Red-labeled siRNA is located in green CD11c-positive cells (arrow).
Mentions: In an attempt to assess how intratracheally administrated siRNA was distributed to the lung, anesthetized mice were administered with Texas Red-labeled non-coding siRNA and then challenged with OVA. One hour and 12 hours after the challenge, frozen sections were made and stained with FITC-labeled anti-CD11c mAb to detect DCs [15]. One hour after the OVA challenge, Red-labeled siRNA was detected on the airway epithelial layer in a band-like deposition pattern. Green-labeled DCs were clustered around the deposited siRNA (Figure 3A). The immunofluorescence of CD11c+cells in proximity of the red siRNA signal was observed in several sites per one section, mainly on the airway epithelial layer. Due to its characteristic distribution, this was unlikely a sectioning/deparaffinizing/staining artifact. Twelve hours after the challenge, the red dots of siRNA were distributed beneath the airway basement membrane and submucosal DCs, indicating that siRNA successfully went through the epithelial barrier (Figure 3B,C). Yellow fluorescence was observed in several sites, suggestive of siRNA taken up by DCs, although a mere topological stratification of fluorescence might not be excluded.Figure 3

Bottom Line: However, it remains uncertain whether this pathway also has an essential role in the effector phase.These results suggest that local administration of CD86 siRNA during the effector phase ameliorates lines of asthma phenotypes.Targeting airway dendritic cells with siRNA suppresses airway inflammation and hyperresponsiveness in an experimental model of allergic asthma.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Diseases of the Chest, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan. yukari-a@kokyu.med.kyushu-u.ac.jp.

ABSTRACT

Background: CD86-CD28 interaction has been suggested as the principal costimulatory pathway for the activation and differentiation of naïve T cells in allergic inflammation. However, it remains uncertain whether this pathway also has an essential role in the effector phase. We sought to determine the contribution of CD86 on dendritic cells in the reactivation of allergen-specific Th2 cells.

Methods: We investigated the effects of the downregulation of CD86 by short interfering RNAs (siRNAs) on Th2 cytokine production in the effector phase in vitro and on asthma phenotypes in ovalbumin (OVA)-sensitized and -challenged mice.

Results: Treatment of bone marrow-derived dendritic cells (BMDCs) with CD86 siRNA attenuated LPS-induced upregulation of CD86. CD86 siRNA treatment impaired BMDCs' ability to activate OVA-specific Th2 cells. Intratracheal administration of CD86 siRNA during OVA challenge downregulated CD86 expression in the airway mucosa. CD86 siRNA treatment ameliorated OVA-induced airway eosinophilia, airway hyperresponsiveness, and the elevations of OVA-specific IgE in the sera and IL-5, IL-13, and CCL17 in the bronchoalveolar lavage fluid, but not the goblet cell hyperplasia.

Conclusion: These results suggest that local administration of CD86 siRNA during the effector phase ameliorates lines of asthma phenotypes. Targeting airway dendritic cells with siRNA suppresses airway inflammation and hyperresponsiveness in an experimental model of allergic asthma.

Show MeSH
Related in: MedlinePlus