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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

Lack of effect of treatment with CD86 siRNA on mucus production in OVA-challenged mice. (A) Lungs were obtained 36 hours after the last OVA challenge and inflated in formalin. Sections were stained with AB/PAS to identify mucus-containing cells. Representative sections from naïve, control siRNA, and CD86 siRNA-treated mice are shown. (B) Semi-quantitative analysis of the abundance of PAS-positive cells. The numeric scores for the abundance of PAS-positive, mucus-containing cells in each airway were determined to be as follows: 0, <5% PAS-positive cells; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, >75%. (C) MUC5AC expression in mouse whole lung was analyzed by real-time PCR. The relative levels of the MUC5AC transcripts were presented as fold increase over baseline values. β-actin was taken as a house-keeping gene. All data mean ± SEM of 6–10 mice per group. n.s. means statistically not significant.
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Fig6: Lack of effect of treatment with CD86 siRNA on mucus production in OVA-challenged mice. (A) Lungs were obtained 36 hours after the last OVA challenge and inflated in formalin. Sections were stained with AB/PAS to identify mucus-containing cells. Representative sections from naïve, control siRNA, and CD86 siRNA-treated mice are shown. (B) Semi-quantitative analysis of the abundance of PAS-positive cells. The numeric scores for the abundance of PAS-positive, mucus-containing cells in each airway were determined to be as follows: 0, <5% PAS-positive cells; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, >75%. (C) MUC5AC expression in mouse whole lung was analyzed by real-time PCR. The relative levels of the MUC5AC transcripts were presented as fold increase over baseline values. β-actin was taken as a house-keeping gene. All data mean ± SEM of 6–10 mice per group. n.s. means statistically not significant.

Mentions: To examine the effects of pulmonary CD86 inhibition on asthma phenotypes, CD86 siRNA was administrated one hour before each OVA challenge (Figures 4 and 5). Thirty-six hours after the last OVA challenge, airway hyperresponsiveness to acetylcholine aerosol was measured. Allergen-induced airway hyperresponsiveness in CD86 siRNA-treated mice was significantly lower than that in control siRNA-treated mice [P =0.0432]. The number of eosinophils in BAL fluid was significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [P =0.0117]. The concentrations of IL-5, IL-13, and CCL17 (TARC [thymus- and activation-regulated chemokine], in the old nomenclature) in BAL fluid were significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [IL-4, P = 0.0101; IL-5, P = 0.0339; IL-13, P =0.0301; CCL17/TARC, P = 0.0479]. The concentration of OVA-specific IgE in the serum was significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [P =0.0425]. However, the goblet cell hyperplasia and MUC5AC gene expression were not significantly different between CD86 siRNA-treated mice and control siRNA-treated mice (Figure 6). These results suggest that intratracheal administration of CD86 siRNA effectively ameliorates lines of asthma phenotypes, except for goblet cell hyperplasia and mucus hyper-secretion, in a murine model of allergic asthma. In addition, we confirmed that there was no elevation of IL-6 and IFN-β in the serum (data not shown) after CD86 siRNA or control siRNA administration, which eliminated the possibility that control siRNA or CD86 siRNA administration induced nonspecific inflammation.Figure 5


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)

Lack of effect of treatment with CD86 siRNA on mucus production in OVA-challenged mice. (A) Lungs were obtained 36 hours after the last OVA challenge and inflated in formalin. Sections were stained with AB/PAS to identify mucus-containing cells. Representative sections from naïve, control siRNA, and CD86 siRNA-treated mice are shown. (B) Semi-quantitative analysis of the abundance of PAS-positive cells. The numeric scores for the abundance of PAS-positive, mucus-containing cells in each airway were determined to be as follows: 0, <5% PAS-positive cells; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, >75%. (C) MUC5AC expression in mouse whole lung was analyzed by real-time PCR. The relative levels of the MUC5AC transcripts were presented as fold increase over baseline values. β-actin was taken as a house-keeping gene. All data mean ± SEM of 6–10 mice per group. n.s. means statistically not significant.
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Related In: Results  -  Collection

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Fig6: Lack of effect of treatment with CD86 siRNA on mucus production in OVA-challenged mice. (A) Lungs were obtained 36 hours after the last OVA challenge and inflated in formalin. Sections were stained with AB/PAS to identify mucus-containing cells. Representative sections from naïve, control siRNA, and CD86 siRNA-treated mice are shown. (B) Semi-quantitative analysis of the abundance of PAS-positive cells. The numeric scores for the abundance of PAS-positive, mucus-containing cells in each airway were determined to be as follows: 0, <5% PAS-positive cells; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, >75%. (C) MUC5AC expression in mouse whole lung was analyzed by real-time PCR. The relative levels of the MUC5AC transcripts were presented as fold increase over baseline values. β-actin was taken as a house-keeping gene. All data mean ± SEM of 6–10 mice per group. n.s. means statistically not significant.
Mentions: To examine the effects of pulmonary CD86 inhibition on asthma phenotypes, CD86 siRNA was administrated one hour before each OVA challenge (Figures 4 and 5). Thirty-six hours after the last OVA challenge, airway hyperresponsiveness to acetylcholine aerosol was measured. Allergen-induced airway hyperresponsiveness in CD86 siRNA-treated mice was significantly lower than that in control siRNA-treated mice [P =0.0432]. The number of eosinophils in BAL fluid was significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [P =0.0117]. The concentrations of IL-5, IL-13, and CCL17 (TARC [thymus- and activation-regulated chemokine], in the old nomenclature) in BAL fluid were significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [IL-4, P = 0.0101; IL-5, P = 0.0339; IL-13, P =0.0301; CCL17/TARC, P = 0.0479]. The concentration of OVA-specific IgE in the serum was significantly lower in CD86 siRNA-treated mice than in control siRNA-treated mice [P =0.0425]. However, the goblet cell hyperplasia and MUC5AC gene expression were not significantly different between CD86 siRNA-treated mice and control siRNA-treated mice (Figure 6). These results suggest that intratracheal administration of CD86 siRNA effectively ameliorates lines of asthma phenotypes, except for goblet cell hyperplasia and mucus hyper-secretion, in a murine model of allergic asthma. In addition, we confirmed that there was no elevation of IL-6 and IFN-β in the serum (data not shown) after CD86 siRNA or control siRNA administration, which eliminated the possibility that control siRNA or CD86 siRNA administration induced nonspecific inflammation.Figure 5

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