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Effects of Olopatadine on the Recovery of Skin Barrier Function and Epidermal Hyperplasia Induced by Skin Barrier Disruption in Mice
S202 Abstracts
793
Effects of Olopatadine on the Recovery of Skin Barrier Function and Epidermal Hyperplasia Induced by Skin Barrier Disruption in Mice T. Amano, T. Takeda, H. Yano, T. Tamura; Kyowa Hakko Kogyo Co., Ltd., Shizuoka, JAPAN. RATIONALE: The skin barrier function in patients with atopic dermatitis is disrupted and prolonged steroid therapy produces epidermal barrier disturbance. We examined here the effect of olopatadine hydrochloride (olopatadine), an antiallergic drug with histamine H1 receptor-antagonistic action, on skin barrier recovery in mice. METHODS: The skin barrier of mice was disrupted by tape stripping. The recovery of skin barrier function was monitored by measurements of transepidermal water loss (TEWL) after barrier disruption. Epidermal hyperplasia was induced by repeated barrier disruption for 7 days. Olopatadine was orally administered once daily from 3 days before barrier disruption. Betamethasone 17-valerate (betamethasone) was topically applied from 3 days before barrier disruption. RESULTS: Tape stripping led to a significant increase in TEWL. TEWL decreased with time after tape stripping and skin barrier function recovered by over 60% within 9 h after tape stripping. The recovery of skin barrier in olopatadine-treated mice was significantly accelerated compared with that in vehicle-treated control. On the other hand, the skin barrier recovery in mice treated with topical betamethasone was significantly delayed compared with that in vehicle-treated mice. Combined treatment of olopatadine with betamethasone ameliorated the delay in barrier recover by topical treatment of betamethasone. In addition, olopatadine significantly prevented the increase in epidermal thickness induced by prolonged barrier disruption. CONCLUSION: These results suggest that systemic administration of olopatadine accelerates the recovery of skin barrier function and ameliorates the adverse effects of topical steroids on the skin barrier recovery.
794
MONDAY
Olopatadine Ameliorates Rat Experimental Cutaneous Inflammation by Improving the Skin Barrier Function T. Tamura, T. Amano, M. Chida, H. Yano; Kyowa Hakko Kogyo Co., Ltd., Shizuoka, JAPAN. RATIONALE: Itching is a characteristic symptom of various forms of dermatitis. Itch-associated scratching damages the skin and increases the inflammation. To investigate the effects of olopatadine hydrochloride (olopatadine), an anti-allergic agent with histamine H1 receptor antagonistic action, on chronic inflammatory dermatitis and impaired skin barrier functions induced by repeated application of oxazolone in rats. METHODS: Oxazolone-sensitized rats were challenged with oxazolone applied to the ear every 3 days. Olopatadine was orally administered once daily. The effects of the drug were quantified by measurements of ear thickness, levels of cytokines in the lesioned ear and the number of scratching episodes. As parameters of skin barrier function, transepidermal water loss (TEWL) and hyaluronic acid (HA) levels in the lesioned ear were measured. The effect of olopatadine on the production of HA by cultured dermal fibroblasts was also measured. RESULTS: Repeated topical application of oxazolone to rat ears induced local inflammation that was exemplified by swelling. In lesioned ear, the amount of IFNg increased at both the protein and mRNA level, but IL-4 levels changed minimally. Olopatadine significantly decreased ear swelling and the number of scratching episodes. The drug also significantly inhibited the increase of IFNg and nerve growth factor production in lesioned ears. Olopatadine significantly inhibited the increase in TEWL and the decrease in HA in lesioned ears. Furthermore, the drug stimulated the production of HA by cultured dermal fibroblasts. CONCLUSIONS: These results suggest that olopatadine suppressed inflammation and scratching not only by inhibiting cytokine production, but also by repairing skin barrier function.
J ALLERGY CLIN IMMUNOL JANUARY 2007
795
IL-13 Receptors in Cutaneous Inflammation M. R. Warrier, A. Gibson, Y. Tabata, M. O. Daines, W. Chen, H. S. Akei, M. E. Rothenberg, G. K. Khurana Hershey; Cincinnati Children’s Hosp. Med. Center, Cincinnati, OH. RATIONALE: IL-13 is a central cytokine in allergic inflammation and is expressed in lesions of atopic dermatitis. IL-13 mediates its effects through a complex receptor system that includes IL-13Ra1 and IL-13Ra2. We have previously shown that expression and distribution of IL-13Ra2, but not IL-13Ra1 is altered by Th2 cytokines in keratinocytes in vitro. We extended these observations in vivo using a murine model of atopic dermatitis. METHODS: Mice underwent three one-week epicutaneous patch applications with house dust mite (HDM), ovalbumin or saline interspersed by two-week patch free intervals. Th2 sensitization was determined by measuring serum HDM or OVA specific-IgE and -IgG1. Epidermal thickening was evaluated by H&E staining. Cutaneous expression of IL-13Ra1 and IL-13Ra2 was measured by quantitative PCR. ELISA was used to measure IL-13Ra2 protein in skin lysates. RESULTS: HDM treated mice were specifically sensitized and demonstrated visible cutaneous inflammation as well as significant epidermal thickening versus saline challenged mice (60 mm 610 vs 23 mm 61.5, p 5 0.005). HDM challenged skin also demonstrated an increase in mRNA for IL-13Ra1 (2.7 fold increase, p 5 0.004) and more significantly IL-13Ra2 (5 fold increase, p < 0.003). Additionally, epicutaneous HDM induced IL-13Ra2 protein in lesional skin greater than 2 fold (p 5 0.003). Similar increases were observed in ovalbumin treated mice. CONCLUSION: Alterations in the expression and distribution of IL-13 receptors, specifically IL-13Ra2, occurs during cutaneous Th2 inflammation and may play a role in the pathogenesis of atopic dermatitis. Funding: NIH
796
Overexpression of Interleukin 13 (IL-13) in the Skin Induces Dermatitis and Pulmonary Inflammation and Airway Hyperresponsiveness to Allergen Challenge T. Zheng, S. Oh, M. Oh, M. Wang, A. Myers, J. Schroeder, Z. Zhu; Johns Hopkins University, Division of Allergy & Clinical Immunology, Baltimore, MD. RATIONALE: Th2 inflammation is a cornerstone in the pathogenesis of asthma and atopic dermatitis (AD). AD is often the first step in ‘‘the atopic march’’ that results in asthma. However, the role of IL-13 in the development of AD and in the relationship between AD and asthma has not been well defined. METHODS: We developed an inducible overexpression transgenic model of AD in which IL-13 can be selectively activated using Tyr promoter in the skin. We used this system to determine if IL-13 induces AD and if this leads to an asthma phenotype. RESULTS: Induction of IL-13 in the skin caused pruritic inflammatory dermatitis characterized by acanthosis, spongiosis, and dermal and perivascular infiltrates dominated by mononuclear cells, CD41 cells, eosinophils and mast cells. After stimulation with anti-CD3 and anti-CD28, CD41 cells from peripheral lymph nodes and spleens of the transgene (1) mice produced significantly higher levels of IL-4, but not IFN-g. The levels of serum IgE, IgG1, but not IgG2a, and skin total histamine in transgene (1) mice were significantly higher than those in transgene (-) mice. In addition, IL-13-induced dermatitis was associated with increased lung inflammation, mucus secretion and airway hypersensitivity after a single dose of Ova challenge. CONCLUSIONS: These studies demonstrate that overexpression of IL13 in the skin causes a pruritic dermatitis with the characteristics closely resembling human AD. This IL-13-induced AD is also associated with a systemic Th2-driven immune response and with increased susceptibility to aeroallergen challenge. IL-13 may be an important mediator in the pathogenesis of AD and in atopic march. Funding: NIH/NIAID
793
Effects of Olopatadine on the Recovery of Skin Barrier Function and Epidermal Hyperplasia Induced by Skin Barrier Disruption in Mice T. Amano, T. Takeda, H. Yano, T. Tamura; Kyowa Hakko Kogyo Co., Ltd., Shizuoka, JAPAN. RATIONALE: The skin barrier function in patients with atopic dermatitis is disrupted and prolonged steroid therapy produces epidermal barrier disturbance. We examined here the effect of olopatadine hydrochloride (olopatadine), an antiallergic drug with histamine H1 receptor-antagonistic action, on skin barrier recovery in mice. METHODS: The skin barrier of mice was disrupted by tape stripping. The recovery of skin barrier function was monitored by measurements of transepidermal water loss (TEWL) after barrier disruption. Epidermal hyperplasia was induced by repeated barrier disruption for 7 days. Olopatadine was orally administered once daily from 3 days before barrier disruption. Betamethasone 17-valerate (betamethasone) was topically applied from 3 days before barrier disruption. RESULTS: Tape stripping led to a significant increase in TEWL. TEWL decreased with time after tape stripping and skin barrier function recovered by over 60% within 9 h after tape stripping. The recovery of skin barrier in olopatadine-treated mice was significantly accelerated compared with that in vehicle-treated control. On the other hand, the skin barrier recovery in mice treated with topical betamethasone was significantly delayed compared with that in vehicle-treated mice. Combined treatment of olopatadine with betamethasone ameliorated the delay in barrier recover by topical treatment of betamethasone. In addition, olopatadine significantly prevented the increase in epidermal thickness induced by prolonged barrier disruption. CONCLUSION: These results suggest that systemic administration of olopatadine accelerates the recovery of skin barrier function and ameliorates the adverse effects of topical steroids on the skin barrier recovery.
794
MONDAY
Olopatadine Ameliorates Rat Experimental Cutaneous Inflammation by Improving the Skin Barrier Function T. Tamura, T. Amano, M. Chida, H. Yano; Kyowa Hakko Kogyo Co., Ltd., Shizuoka, JAPAN. RATIONALE: Itching is a characteristic symptom of various forms of dermatitis. Itch-associated scratching damages the skin and increases the inflammation. To investigate the effects of olopatadine hydrochloride (olopatadine), an anti-allergic agent with histamine H1 receptor antagonistic action, on chronic inflammatory dermatitis and impaired skin barrier functions induced by repeated application of oxazolone in rats. METHODS: Oxazolone-sensitized rats were challenged with oxazolone applied to the ear every 3 days. Olopatadine was orally administered once daily. The effects of the drug were quantified by measurements of ear thickness, levels of cytokines in the lesioned ear and the number of scratching episodes. As parameters of skin barrier function, transepidermal water loss (TEWL) and hyaluronic acid (HA) levels in the lesioned ear were measured. The effect of olopatadine on the production of HA by cultured dermal fibroblasts was also measured. RESULTS: Repeated topical application of oxazolone to rat ears induced local inflammation that was exemplified by swelling. In lesioned ear, the amount of IFNg increased at both the protein and mRNA level, but IL-4 levels changed minimally. Olopatadine significantly decreased ear swelling and the number of scratching episodes. The drug also significantly inhibited the increase of IFNg and nerve growth factor production in lesioned ears. Olopatadine significantly inhibited the increase in TEWL and the decrease in HA in lesioned ears. Furthermore, the drug stimulated the production of HA by cultured dermal fibroblasts. CONCLUSIONS: These results suggest that olopatadine suppressed inflammation and scratching not only by inhibiting cytokine production, but also by repairing skin barrier function.
J ALLERGY CLIN IMMUNOL JANUARY 2007
795
IL-13 Receptors in Cutaneous Inflammation M. R. Warrier, A. Gibson, Y. Tabata, M. O. Daines, W. Chen, H. S. Akei, M. E. Rothenberg, G. K. Khurana Hershey; Cincinnati Children’s Hosp. Med. Center, Cincinnati, OH. RATIONALE: IL-13 is a central cytokine in allergic inflammation and is expressed in lesions of atopic dermatitis. IL-13 mediates its effects through a complex receptor system that includes IL-13Ra1 and IL-13Ra2. We have previously shown that expression and distribution of IL-13Ra2, but not IL-13Ra1 is altered by Th2 cytokines in keratinocytes in vitro. We extended these observations in vivo using a murine model of atopic dermatitis. METHODS: Mice underwent three one-week epicutaneous patch applications with house dust mite (HDM), ovalbumin or saline interspersed by two-week patch free intervals. Th2 sensitization was determined by measuring serum HDM or OVA specific-IgE and -IgG1. Epidermal thickening was evaluated by H&E staining. Cutaneous expression of IL-13Ra1 and IL-13Ra2 was measured by quantitative PCR. ELISA was used to measure IL-13Ra2 protein in skin lysates. RESULTS: HDM treated mice were specifically sensitized and demonstrated visible cutaneous inflammation as well as significant epidermal thickening versus saline challenged mice (60 mm 610 vs 23 mm 61.5, p 5 0.005). HDM challenged skin also demonstrated an increase in mRNA for IL-13Ra1 (2.7 fold increase, p 5 0.004) and more significantly IL-13Ra2 (5 fold increase, p < 0.003). Additionally, epicutaneous HDM induced IL-13Ra2 protein in lesional skin greater than 2 fold (p 5 0.003). Similar increases were observed in ovalbumin treated mice. CONCLUSION: Alterations in the expression and distribution of IL-13 receptors, specifically IL-13Ra2, occurs during cutaneous Th2 inflammation and may play a role in the pathogenesis of atopic dermatitis. Funding: NIH
796
Overexpression of Interleukin 13 (IL-13) in the Skin Induces Dermatitis and Pulmonary Inflammation and Airway Hyperresponsiveness to Allergen Challenge T. Zheng, S. Oh, M. Oh, M. Wang, A. Myers, J. Schroeder, Z. Zhu; Johns Hopkins University, Division of Allergy & Clinical Immunology, Baltimore, MD. RATIONALE: Th2 inflammation is a cornerstone in the pathogenesis of asthma and atopic dermatitis (AD). AD is often the first step in ‘‘the atopic march’’ that results in asthma. However, the role of IL-13 in the development of AD and in the relationship between AD and asthma has not been well defined. METHODS: We developed an inducible overexpression transgenic model of AD in which IL-13 can be selectively activated using Tyr promoter in the skin. We used this system to determine if IL-13 induces AD and if this leads to an asthma phenotype. RESULTS: Induction of IL-13 in the skin caused pruritic inflammatory dermatitis characterized by acanthosis, spongiosis, and dermal and perivascular infiltrates dominated by mononuclear cells, CD41 cells, eosinophils and mast cells. After stimulation with anti-CD3 and anti-CD28, CD41 cells from peripheral lymph nodes and spleens of the transgene (1) mice produced significantly higher levels of IL-4, but not IFN-g. The levels of serum IgE, IgG1, but not IgG2a, and skin total histamine in transgene (1) mice were significantly higher than those in transgene (-) mice. In addition, IL-13-induced dermatitis was associated with increased lung inflammation, mucus secretion and airway hypersensitivity after a single dose of Ova challenge. CONCLUSIONS: These studies demonstrate that overexpression of IL13 in the skin causes a pruritic dermatitis with the characteristics closely resembling human AD. This IL-13-induced AD is also associated with a systemic Th2-driven immune response and with increased susceptibility to aeroallergen challenge. IL-13 may be an important mediator in the pathogenesis of AD and in atopic march. Funding: NIH/NIAID