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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

Th2 cytokine levels in culture supernatant were analyzed by ELISA. (A) DO11.10 spleen cells were primed with pOVA and APCs in the presence of rIL-4, anti-IL-12 antibody, CD28, and IL-2. Seven days later, CD4-positive cells were purified using MACS and used as effector T cells. CD86 siRNA-transfected BMDCs were pulsed with pOVA and stimulated with LPS and then cultured with effector T cells. (B) Coculture supernatants were collected 48 hours later, and cytokine levels were measured by ELISA (n = 4; mean ± SEM). * indicates statistically significant (P <0.05) difference between the indicated two groups.
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Fig2: Th2 cytokine levels in culture supernatant were analyzed by ELISA. (A) DO11.10 spleen cells were primed with pOVA and APCs in the presence of rIL-4, anti-IL-12 antibody, CD28, and IL-2. Seven days later, CD4-positive cells were purified using MACS and used as effector T cells. CD86 siRNA-transfected BMDCs were pulsed with pOVA and stimulated with LPS and then cultured with effector T cells. (B) Coculture supernatants were collected 48 hours later, and cytokine levels were measured by ELISA (n = 4; mean ± SEM). * indicates statistically significant (P <0.05) difference between the indicated two groups.

Mentions: Although many previous studies have shown an essential role of CD86 on APCs in the antigen priming of naïve T cells and subsequent Th2 commitment, there have been no consistent results for the role of CD86 in the reactivation of antigen-specific Th2 cells. OVA-specific Th2 cells were induced by coculture of CD4+T cells purified from the spleens of DO11.10 mice with APCs from the spleens of naïve BALB/c mice in the presence of pOVA, recombinant IL-4, neutralizing anti-IL-12 mAb, and agonistic anti-CD28 mAb, and followed by expansion with recombinant IL-2 (Figure 2A). Th2 specificity was confirmed by selective production of IL-4, IL-5, and IL-13, but to a smaller extent, IFN-γ, which is induced by coculture of the cells with pOVA-loaded and LPS-stimulated BMDCs (referred to as pOVA/LPS-BMDCs). The contents of IL-4, IL-5, and IL-13 in the culture supernatants from coculture of the cells with CD86 siRNA-treated pOVA/LPS-BMDCs were significantly lower than those from coculture of the cells with control siRNA-treated pOVA/LPS-BMDCs [IL-4, P =0.0209; IL-5, P = 0.0209; IL-13, P =0.0209] (Figure 2B). When BMDCs were loaded with pOVA but not stimulated with LPS, almost no Th2 cytokine production was detected (data not shown). These results suggest that the reactivation of murine antigen-specific Th2 cells in vitro is partially dependent on CD86 on APCs.Figure 2


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)

Th2 cytokine levels in culture supernatant were analyzed by ELISA. (A) DO11.10 spleen cells were primed with pOVA and APCs in the presence of rIL-4, anti-IL-12 antibody, CD28, and IL-2. Seven days later, CD4-positive cells were purified using MACS and used as effector T cells. CD86 siRNA-transfected BMDCs were pulsed with pOVA and stimulated with LPS and then cultured with effector T cells. (B) Coculture supernatants were collected 48 hours later, and cytokine levels were measured by ELISA (n = 4; mean ± SEM). * 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

Fig2: Th2 cytokine levels in culture supernatant were analyzed by ELISA. (A) DO11.10 spleen cells were primed with pOVA and APCs in the presence of rIL-4, anti-IL-12 antibody, CD28, and IL-2. Seven days later, CD4-positive cells were purified using MACS and used as effector T cells. CD86 siRNA-transfected BMDCs were pulsed with pOVA and stimulated with LPS and then cultured with effector T cells. (B) Coculture supernatants were collected 48 hours later, and cytokine levels were measured by ELISA (n = 4; mean ± SEM). * indicates statistically significant (P <0.05) difference between the indicated two groups.
Mentions: Although many previous studies have shown an essential role of CD86 on APCs in the antigen priming of naïve T cells and subsequent Th2 commitment, there have been no consistent results for the role of CD86 in the reactivation of antigen-specific Th2 cells. OVA-specific Th2 cells were induced by coculture of CD4+T cells purified from the spleens of DO11.10 mice with APCs from the spleens of naïve BALB/c mice in the presence of pOVA, recombinant IL-4, neutralizing anti-IL-12 mAb, and agonistic anti-CD28 mAb, and followed by expansion with recombinant IL-2 (Figure 2A). Th2 specificity was confirmed by selective production of IL-4, IL-5, and IL-13, but to a smaller extent, IFN-γ, which is induced by coculture of the cells with pOVA-loaded and LPS-stimulated BMDCs (referred to as pOVA/LPS-BMDCs). The contents of IL-4, IL-5, and IL-13 in the culture supernatants from coculture of the cells with CD86 siRNA-treated pOVA/LPS-BMDCs were significantly lower than those from coculture of the cells with control siRNA-treated pOVA/LPS-BMDCs [IL-4, P =0.0209; IL-5, P = 0.0209; IL-13, P =0.0209] (Figure 2B). When BMDCs were loaded with pOVA but not stimulated with LPS, almost no Th2 cytokine production was detected (data not shown). These results suggest that the reactivation of murine antigen-specific Th2 cells in vitro is partially dependent on CD86 on APCs.Figure 2

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