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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

Effect of siRNA treatment on CD86 positive cells around airway. Sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. Naïve mice served as negative control. (A) Lung sections from naïve, CD86 siRNA-treated or control siRNA-treated mice stained with FITC-conjugated anti-CD86 mAb. Bar, 200 μm for upper panel; 100 μm for lower panel. The upper panel and lower panel were obtained from different mice (B) Micrographs were quantitatively analyzed for the presence of FITC-anti-CD86 signals per unit area with image analysis software. All data mean ± SEM of 3–4 mice per group. * indicates statistically significant (P <0.05) difference between the indicated two groups.
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Fig4: Effect of siRNA treatment on CD86 positive cells around airway. Sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. Naïve mice served as negative control. (A) Lung sections from naïve, CD86 siRNA-treated or control siRNA-treated mice stained with FITC-conjugated anti-CD86 mAb. Bar, 200 μm for upper panel; 100 μm for lower panel. The upper panel and lower panel were obtained from different mice (B) Micrographs were quantitatively analyzed for the presence of FITC-anti-CD86 signals per unit area with image analysis software. All data mean ± SEM of 3–4 mice per group. * indicates statistically significant (P <0.05) difference between the indicated two groups.

Mentions: Next, for evaluation of the interfering efficacy, sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. CD86-positive cells were detected by biotin-conjugated anti-CD86 mAb followed by streptavidin-conjugated FITC and assessed semi-quantitatively as CD86-positive areas by image analysis. As shown in Figure 4, in the sections of naïve mice, there were scarce CD86-positive cells in the airway mucosa, although the area of CD86-positive cells was markedly increased in control siRNA-treated and OVA-challenged mice. CD86-expressing cells were detected in dense clusters beneath the airway epithelium. The upregulation of CD86-positive cells was significantly suppressed in the CD86 siRNA-treated mice [P =0.0036]. These results demonstrate that local administration of siRNA ameliorates the allergen-induced upregulation of CD86 expression on the airway DCs.Figure 4


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)

Effect of siRNA treatment on CD86 positive cells around airway. Sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. Naïve mice served as negative control. (A) Lung sections from naïve, CD86 siRNA-treated or control siRNA-treated mice stained with FITC-conjugated anti-CD86 mAb. Bar, 200 μm for upper panel; 100 μm for lower panel. The upper panel and lower panel were obtained from different mice (B) Micrographs were quantitatively analyzed for the presence of FITC-anti-CD86 signals per unit area with image analysis software. All data mean ± SEM of 3–4 mice per group. * indicates statistically significant (P <0.05) difference between the indicated two groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: Effect of siRNA treatment on CD86 positive cells around airway. Sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. Naïve mice served as negative control. (A) Lung sections from naïve, CD86 siRNA-treated or control siRNA-treated mice stained with FITC-conjugated anti-CD86 mAb. Bar, 200 μm for upper panel; 100 μm for lower panel. The upper panel and lower panel were obtained from different mice (B) Micrographs were quantitatively analyzed for the presence of FITC-anti-CD86 signals per unit area with image analysis software. All data mean ± SEM of 3–4 mice per group. * indicates statistically significant (P <0.05) difference between the indicated two groups.
Mentions: Next, for evaluation of the interfering efficacy, sensitized mice were intratracheally administered with CD86 siRNA or control siRNA and given a single challenge with OVA. Twelve hours after the challenge, the tracheas and lungs were removed and processed for immune-fluorescence study. CD86-positive cells were detected by biotin-conjugated anti-CD86 mAb followed by streptavidin-conjugated FITC and assessed semi-quantitatively as CD86-positive areas by image analysis. As shown in Figure 4, in the sections of naïve mice, there were scarce CD86-positive cells in the airway mucosa, although the area of CD86-positive cells was markedly increased in control siRNA-treated and OVA-challenged mice. CD86-expressing cells were detected in dense clusters beneath the airway epithelium. The upregulation of CD86-positive cells was significantly suppressed in the CD86 siRNA-treated mice [P =0.0036]. These results demonstrate that local administration of siRNA ameliorates the allergen-induced upregulation of CD86 expression on the airway DCs.Figure 4

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