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1060 Fenoterol inhibits the β receptor-independent signaling pathways of superoxide anion generation inhuman neutrophils
S360
Abstracts
1059
Influence of Inhaled Steroids as a Treatment 5n Asthmatic Children and Their Relationship With IgE Sensitization to House Dust Mite and Lolium Perenne G tedn*, SE Soro Lavin*,
J ALLERGY CLIN IMMUNOL JANUARY 2000
E Scheihing*.
C Diazt,
AM Von Chrismar*.
J lnostrozaf
*Austral University of Chile TUFRO Inhaled steroids in the treatment of allergic asthma in children have been useful to control symptoms of the exacerbations of the disease because of their antiinflammatory effects. They can affect cortisol level and growth in children.It is not known if specific sensitization can be modified with inhaled steroids as a treatment. The aim of our study was to compare IgE levels before and after 6 weeks of treatment with budesonide (17 patients), fluticasone ( 18 patients) and beclometasone (20 patients). Mean age was 9 years old ( 5 to 15 years old).A blood sample was obtained in all patients. Unicap system was used to process specific IgE for House Dust Mite and Lolium Perenne. Statistical analysis was done with parametric and non parametric tests. The analysis done between before and after 6 weeks of treatment showed a p-value 0.0354 ( t-test), 0.0549 (Wilcoxon test) for House Dust Mite and 0.0093 ( t-test), 0.0028 (Wilcoxon test) for Lilium Perenne. Therefore, we conclude that inhaled steroids can modified the sensitization to both allergens.
1060
ly, all three drugs did not show the 02-scavenging effect. These results indicate that all three drugs have 0 receptor-dependent inhibitory mechanism on 02- generation. However, fenoterol also may have a p receptor-independent signaling pathway of 02- generation in human neutrophils.
Fenoterol Inhibits the p Receptor-independent Signaling Pathways of Superoxide Anion Generation in Human Neutrophils A Tachibana*. M Kate*. ZN Mirza*. H Kimurat. K Tokuyama*, A Morikawa* *Department of Pediatrics, Gunma University School of Medicine tGunma University School of Medicine and Gunma Prefectural Institute of Public Health and Environmental Sciences, Maebashi, Gunma, Japan Oxygen radicals derived from human inflammatory cells and tissue damages contribute to the pathophysiology of bronchail asthma. We previously reported that some p agonists inhibited the superoxide anion (02-) generation in human neutrophils. Here, we investigated effect of the different kinds of p agonists such as fenoterol as a known full 0 agonist and salbutamol or procaterol as a known partial p agonist on 0,. generation in neutrophils by two stimuli such as N-formyl-methionyl phenylalanine, FMLP (a stimulator which bind to G protein-coupled receptor) or phorbol miristate acetate, PMA (a direct activator of protein kinase C, PKC). The 02- generation was assayed by a highly sensitive chemiluminescence method using a Cypridina luciferin analog as the amplifier. We found that fenoterol inhibited both FMLP- and PMA-induced 0,. generation in a dose-dependent manner. In contrast, salbutamol or procaterol inhibited only FMLP-induced 0,. generation. Secondarily. to confirm whether these inhibitory effect of p agonists are dependent on p receptor, we tested the effect of propranolol, a 0 blocker, on stimuli-induced O,- generation inhibited by p agonist. As a result, the inhibitory effect of salbutamol or procaterol on FMLP-induced 0,. generation was completely reversed in the pretreatment of propranolol. On the other hand, inhibitory effect of fenoterol on FMLP-induced 0,. generation was partially reversed in the pretreatment of propranolol. Furthermore, the inhibitory effect of fenoterol on PMA-induced O?- generation was not reversed in the pretreatment of propranolol. Final-
1061
Tyrosine Kinase Inhibitor 2,4,6-Trihydroxy-ap-methoxyphenylacetophenone (Compound D-58) Is a Potent Inhibitor of Allergic and Inflammatory Reactions Fatih lJckun*d Rama Malaviyad. Shgi-Tai Jan$i De-Min Zhud Ravi Malaviya# *Drug Discovery Program tImmunology SDepartment of Allergy and Inflammatory Diseases OChemistry IHughes Institute, St Paul, MN Proinflammatory mediators such as histamine, arachidonic acid metabolites and a number of cytokines have been implicated in the pathogenesis of a number of acute and chronic inflammatory conditions such as allergy, asthma, arthritis, psoriasis, and skin sunburn. The release of these inflammatory agents is mediated by a cascade of intracellular signaling events which include tyrosine phosphorylation of several cytosolic and membrane proteins. We investigated the anti-allergic and anti-inflammatory effects of a novel compound, 2,4,6-Trihydroxy--p-methoxyphenylacetophenone (Compound D-58) which inhibits tyrosine kinases SYK and BTK. Compound D-58 inhibited IgE receptor/FcERI-mediated mast cell degranulation and leukotriene (LT)C, release and UVBinduced human keratinocyte prostagandin (PG)E, release in a concentration-dependent fashion. Notably, compound D-58 prevented the mast cell mediator-induced vascular hyperpermeability in an in vivo murine model of passive cutaneous anaphylaxis as measured by the prevention of extravasation of systemically administered Evan’s blue dye. Further, compound D-58 treatment effectively inhibited the development of skin edema in the skin of UVB irradiated mice and release of PGE, and LTBJ and neutrophil influx in an airpouch model of inflammation. In conclusion, our results indicate that compound D-58 (2,4,6-Trihydroxy a-p-methoxyphenylacetophenone) has potent anti-allergic and anti-inflammatory properties. Therefore, further development of compound D-58 may provide the basis for new and effective treatment programs for severe allergic as well as inflammatory disorders.
1062
D-AKAP2 INHIBITS Gs alpha-INDUCED CAMP GENERATION IN HEK293 CELLS Andrei Trosrel. Kirk Druey Laboratory of Allergic Diseases, NIAID, NIH, Rockville, MD Regulators of G-protein Signaling (RGS) proteins inhibit signaling pathways employing G proteins of the Gi, Gq and G13 subfamilies of heterotrimeric G proteins by acting as GTPase activating proteins (GAPS). To date no RGS proteins have been shown to act as GAPS for Gs alpha. We have cloned and expressed a novel protein with a highly divergent RGS domain, D-AKAP2, which also contains a novel protein kinase A anchoring motif. No binding of D-AKAP2 to any G protein was detected. Expression of full length D-AKAP2 or its RGS domain in human embryonic kidney (HEK) 293 cells did not result in attenuation of signaling output through Gi or Gq-linked receptors nor did it affect stimu-
Abstracts
1059
Influence of Inhaled Steroids as a Treatment 5n Asthmatic Children and Their Relationship With IgE Sensitization to House Dust Mite and Lolium Perenne G tedn*, SE Soro Lavin*,
J ALLERGY CLIN IMMUNOL JANUARY 2000
E Scheihing*.
C Diazt,
AM Von Chrismar*.
J lnostrozaf
*Austral University of Chile TUFRO Inhaled steroids in the treatment of allergic asthma in children have been useful to control symptoms of the exacerbations of the disease because of their antiinflammatory effects. They can affect cortisol level and growth in children.It is not known if specific sensitization can be modified with inhaled steroids as a treatment. The aim of our study was to compare IgE levels before and after 6 weeks of treatment with budesonide (17 patients), fluticasone ( 18 patients) and beclometasone (20 patients). Mean age was 9 years old ( 5 to 15 years old).A blood sample was obtained in all patients. Unicap system was used to process specific IgE for House Dust Mite and Lolium Perenne. Statistical analysis was done with parametric and non parametric tests. The analysis done between before and after 6 weeks of treatment showed a p-value 0.0354 ( t-test), 0.0549 (Wilcoxon test) for House Dust Mite and 0.0093 ( t-test), 0.0028 (Wilcoxon test) for Lilium Perenne. Therefore, we conclude that inhaled steroids can modified the sensitization to both allergens.
1060
ly, all three drugs did not show the 02-scavenging effect. These results indicate that all three drugs have 0 receptor-dependent inhibitory mechanism on 02- generation. However, fenoterol also may have a p receptor-independent signaling pathway of 02- generation in human neutrophils.
Fenoterol Inhibits the p Receptor-independent Signaling Pathways of Superoxide Anion Generation in Human Neutrophils A Tachibana*. M Kate*. ZN Mirza*. H Kimurat. K Tokuyama*, A Morikawa* *Department of Pediatrics, Gunma University School of Medicine tGunma University School of Medicine and Gunma Prefectural Institute of Public Health and Environmental Sciences, Maebashi, Gunma, Japan Oxygen radicals derived from human inflammatory cells and tissue damages contribute to the pathophysiology of bronchail asthma. We previously reported that some p agonists inhibited the superoxide anion (02-) generation in human neutrophils. Here, we investigated effect of the different kinds of p agonists such as fenoterol as a known full 0 agonist and salbutamol or procaterol as a known partial p agonist on 0,. generation in neutrophils by two stimuli such as N-formyl-methionyl phenylalanine, FMLP (a stimulator which bind to G protein-coupled receptor) or phorbol miristate acetate, PMA (a direct activator of protein kinase C, PKC). The 02- generation was assayed by a highly sensitive chemiluminescence method using a Cypridina luciferin analog as the amplifier. We found that fenoterol inhibited both FMLP- and PMA-induced 0,. generation in a dose-dependent manner. In contrast, salbutamol or procaterol inhibited only FMLP-induced 0,. generation. Secondarily. to confirm whether these inhibitory effect of p agonists are dependent on p receptor, we tested the effect of propranolol, a 0 blocker, on stimuli-induced O,- generation inhibited by p agonist. As a result, the inhibitory effect of salbutamol or procaterol on FMLP-induced 0,. generation was completely reversed in the pretreatment of propranolol. On the other hand, inhibitory effect of fenoterol on FMLP-induced 0,. generation was partially reversed in the pretreatment of propranolol. Furthermore, the inhibitory effect of fenoterol on PMA-induced O?- generation was not reversed in the pretreatment of propranolol. Final-
1061
Tyrosine Kinase Inhibitor 2,4,6-Trihydroxy-ap-methoxyphenylacetophenone (Compound D-58) Is a Potent Inhibitor of Allergic and Inflammatory Reactions Fatih lJckun*d Rama Malaviyad. Shgi-Tai Jan$i De-Min Zhud Ravi Malaviya# *Drug Discovery Program tImmunology SDepartment of Allergy and Inflammatory Diseases OChemistry IHughes Institute, St Paul, MN Proinflammatory mediators such as histamine, arachidonic acid metabolites and a number of cytokines have been implicated in the pathogenesis of a number of acute and chronic inflammatory conditions such as allergy, asthma, arthritis, psoriasis, and skin sunburn. The release of these inflammatory agents is mediated by a cascade of intracellular signaling events which include tyrosine phosphorylation of several cytosolic and membrane proteins. We investigated the anti-allergic and anti-inflammatory effects of a novel compound, 2,4,6-Trihydroxy--p-methoxyphenylacetophenone (Compound D-58) which inhibits tyrosine kinases SYK and BTK. Compound D-58 inhibited IgE receptor/FcERI-mediated mast cell degranulation and leukotriene (LT)C, release and UVBinduced human keratinocyte prostagandin (PG)E, release in a concentration-dependent fashion. Notably, compound D-58 prevented the mast cell mediator-induced vascular hyperpermeability in an in vivo murine model of passive cutaneous anaphylaxis as measured by the prevention of extravasation of systemically administered Evan’s blue dye. Further, compound D-58 treatment effectively inhibited the development of skin edema in the skin of UVB irradiated mice and release of PGE, and LTBJ and neutrophil influx in an airpouch model of inflammation. In conclusion, our results indicate that compound D-58 (2,4,6-Trihydroxy a-p-methoxyphenylacetophenone) has potent anti-allergic and anti-inflammatory properties. Therefore, further development of compound D-58 may provide the basis for new and effective treatment programs for severe allergic as well as inflammatory disorders.
1062
D-AKAP2 INHIBITS Gs alpha-INDUCED CAMP GENERATION IN HEK293 CELLS Andrei Trosrel. Kirk Druey Laboratory of Allergic Diseases, NIAID, NIH, Rockville, MD Regulators of G-protein Signaling (RGS) proteins inhibit signaling pathways employing G proteins of the Gi, Gq and G13 subfamilies of heterotrimeric G proteins by acting as GTPase activating proteins (GAPS). To date no RGS proteins have been shown to act as GAPS for Gs alpha. We have cloned and expressed a novel protein with a highly divergent RGS domain, D-AKAP2, which also contains a novel protein kinase A anchoring motif. No binding of D-AKAP2 to any G protein was detected. Expression of full length D-AKAP2 or its RGS domain in human embryonic kidney (HEK) 293 cells did not result in attenuation of signaling output through Gi or Gq-linked receptors nor did it affect stimu-