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Treatment of allergic airway inflammation and hyperresponsiveness by antisense-induced local blockade of GATA-3 expression.

Finotto S, De Sanctis GT, Lehr HA, Herz U, Buerke M, Schipp M, Bartsch B, Atreya R, Schmitt E, Galle PR, Renz H, Neurath MF - J. Exp. Med. (2001)

Bottom Line: However, it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness.Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production.This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, University of Mainz, 55099 Mainz, Germany. sfinotto@rics.bwh.harvard.edu

ABSTRACT
Recent studies in transgenic mice have revealed that expression of a dominant negative form of the transcription factor GATA-3 in T cells can prevent T helper cell type 2 (Th2)-mediated allergic airway inflammation in mice. However, it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness. Here, we analyzed the effects of locally antagonizing GATA-3 function in a murine model of asthma. We could suppress GATA-3 expression in interleukin (IL)-4-producing T cells in vitro and in vivo by an antisense phosphorothioate oligonucleotide overlapping the translation start site of GATA-3, whereas nonsense control oligonucleotides were virtually inactive. In a murine model of asthma associated with allergic pulmonary inflammation and hyperresponsiveness in ovalbumin (OVA)-sensitized mice, local intranasal administration of fluorescein isothiocyanate-labeled GATA-3 antisense oligonucleotides led to DNA uptake in lung cells associated with a reduction of intracellular GATA-3 expression. Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production. Furthermore, treatment with antisense but not nonsense oligonucleotides induced a significant reduction of airway hyperresponsiveness in OVA-sensitized mice to levels comparable to saline-treated control mice, as assessed by both enhanced pause (PenH) responses and pulmonary resistance determined by body plethysmography. These data indicate a critical role for GATA-3 in the effector phase of a murine asthma model and suggest that local delivery of GATA-3 antisense oligonucleotides may be a novel approach for the treatment of airway hyperresponsiveness such as in asthma. This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine.

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Histologic evidence of airway inflammation in OVA-sensitized and control mice; effect of GATA-3 antisense treatment. Lung tissue was analyzed from untreated (C and D) and antisense-treated (E and F) OVA-sensitized mice. In untreated mice, a massive peribronchial infiltration with eosinophils, thickening of the basement membrane, and deepithelialization were seen (see higher magnification in D: ×400). In contrast, after treatment with antisense DNA to GATA-3 an intact bronchial epithelial layer and no eosinophil infiltration were seen (E and F) comparable to DEX treatment (I and K). Lung tissues taken from sham (saline)- sensitized mice (A and B) and OVA-immunized mice treated with nonsense DNA (G and H) are shown as control. Lung sections were stained with hematoxylin and eosin and examined by light microscopy. Original magnifications: (A, C, E, G, and H) ×200; (B, D, F, H, and K) ×400.
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Figure 4: Histologic evidence of airway inflammation in OVA-sensitized and control mice; effect of GATA-3 antisense treatment. Lung tissue was analyzed from untreated (C and D) and antisense-treated (E and F) OVA-sensitized mice. In untreated mice, a massive peribronchial infiltration with eosinophils, thickening of the basement membrane, and deepithelialization were seen (see higher magnification in D: ×400). In contrast, after treatment with antisense DNA to GATA-3 an intact bronchial epithelial layer and no eosinophil infiltration were seen (E and F) comparable to DEX treatment (I and K). Lung tissues taken from sham (saline)- sensitized mice (A and B) and OVA-immunized mice treated with nonsense DNA (G and H) are shown as control. Lung sections were stained with hematoxylin and eosin and examined by light microscopy. Original magnifications: (A, C, E, G, and H) ×200; (B, D, F, H, and K) ×400.

Mentions: To assess the histologic effects of GATA-3 antisense phosphorothioate oligonucleotides on allergen-induced airway inflammation, we analyzed lung tissues 24 h after the last of three sequential OVA challenges (days 25–27) on day 28. In untreated OVA-sensitized mice, an infiltration of the bronchial interstitium with eosinophils was observed (Fig. 4C and Fig. D). Treatment of OVA-sensitized mice with nonsense GATA-3 oligonucleotides had no detectable effects (Fig. 4G and Fig. H). In contrast, no inflammatory signs were seen in OVA-sensitized mice upon treatment with corticosteroids or GATA-3 antisense oligonucleotides (Fig. 4I, Fig. K, Fig. E, and Fig. F). Control mice were treated with saline complexed with alum and rechallenged intranasally with saline; they show an intact airway epithelium and no sign of inflammation (Fig. 4A and Fig. B).


Treatment of allergic airway inflammation and hyperresponsiveness by antisense-induced local blockade of GATA-3 expression.

Finotto S, De Sanctis GT, Lehr HA, Herz U, Buerke M, Schipp M, Bartsch B, Atreya R, Schmitt E, Galle PR, Renz H, Neurath MF - J. Exp. Med. (2001)

Histologic evidence of airway inflammation in OVA-sensitized and control mice; effect of GATA-3 antisense treatment. Lung tissue was analyzed from untreated (C and D) and antisense-treated (E and F) OVA-sensitized mice. In untreated mice, a massive peribronchial infiltration with eosinophils, thickening of the basement membrane, and deepithelialization were seen (see higher magnification in D: ×400). In contrast, after treatment with antisense DNA to GATA-3 an intact bronchial epithelial layer and no eosinophil infiltration were seen (E and F) comparable to DEX treatment (I and K). Lung tissues taken from sham (saline)- sensitized mice (A and B) and OVA-immunized mice treated with nonsense DNA (G and H) are shown as control. Lung sections were stained with hematoxylin and eosin and examined by light microscopy. Original magnifications: (A, C, E, G, and H) ×200; (B, D, F, H, and K) ×400.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2193377&req=5

Figure 4: Histologic evidence of airway inflammation in OVA-sensitized and control mice; effect of GATA-3 antisense treatment. Lung tissue was analyzed from untreated (C and D) and antisense-treated (E and F) OVA-sensitized mice. In untreated mice, a massive peribronchial infiltration with eosinophils, thickening of the basement membrane, and deepithelialization were seen (see higher magnification in D: ×400). In contrast, after treatment with antisense DNA to GATA-3 an intact bronchial epithelial layer and no eosinophil infiltration were seen (E and F) comparable to DEX treatment (I and K). Lung tissues taken from sham (saline)- sensitized mice (A and B) and OVA-immunized mice treated with nonsense DNA (G and H) are shown as control. Lung sections were stained with hematoxylin and eosin and examined by light microscopy. Original magnifications: (A, C, E, G, and H) ×200; (B, D, F, H, and K) ×400.
Mentions: To assess the histologic effects of GATA-3 antisense phosphorothioate oligonucleotides on allergen-induced airway inflammation, we analyzed lung tissues 24 h after the last of three sequential OVA challenges (days 25–27) on day 28. In untreated OVA-sensitized mice, an infiltration of the bronchial interstitium with eosinophils was observed (Fig. 4C and Fig. D). Treatment of OVA-sensitized mice with nonsense GATA-3 oligonucleotides had no detectable effects (Fig. 4G and Fig. H). In contrast, no inflammatory signs were seen in OVA-sensitized mice upon treatment with corticosteroids or GATA-3 antisense oligonucleotides (Fig. 4I, Fig. K, Fig. E, and Fig. F). Control mice were treated with saline complexed with alum and rechallenged intranasally with saline; they show an intact airway epithelium and no sign of inflammation (Fig. 4A and Fig. B).

Bottom Line: However, it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness.Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production.This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology, University of Mainz, 55099 Mainz, Germany. sfinotto@rics.bwh.harvard.edu

ABSTRACT
Recent studies in transgenic mice have revealed that expression of a dominant negative form of the transcription factor GATA-3 in T cells can prevent T helper cell type 2 (Th2)-mediated allergic airway inflammation in mice. However, it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness. Here, we analyzed the effects of locally antagonizing GATA-3 function in a murine model of asthma. We could suppress GATA-3 expression in interleukin (IL)-4-producing T cells in vitro and in vivo by an antisense phosphorothioate oligonucleotide overlapping the translation start site of GATA-3, whereas nonsense control oligonucleotides were virtually inactive. In a murine model of asthma associated with allergic pulmonary inflammation and hyperresponsiveness in ovalbumin (OVA)-sensitized mice, local intranasal administration of fluorescein isothiocyanate-labeled GATA-3 antisense oligonucleotides led to DNA uptake in lung cells associated with a reduction of intracellular GATA-3 expression. Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production. Furthermore, treatment with antisense but not nonsense oligonucleotides induced a significant reduction of airway hyperresponsiveness in OVA-sensitized mice to levels comparable to saline-treated control mice, as assessed by both enhanced pause (PenH) responses and pulmonary resistance determined by body plethysmography. These data indicate a critical role for GATA-3 in the effector phase of a murine asthma model and suggest that local delivery of GATA-3 antisense oligonucleotides may be a novel approach for the treatment of airway hyperresponsiveness such as in asthma. This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine.

Show MeSH
Related in: MedlinePlus