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792 Therapeutic potential of specific chymase inhibitor in atopic dermatitis
Abstracts
J ALLERGY CLIN IMMUNOL VOLUME 104. NUMBER 1, PART 2
studies in these patients as diagnosis of SCID can treatment as well as assist in genetic counseling. 790
791
dictate
priority
were observed at 6 hrs (data not shown). Cells positive for IL-4 or IL-5 mRNA (by in situ hybridization) were less frequent in Ag+Candida sites than in Ag sites possibly due to paracrine suppressive effects on Th2 responses by the Thl responses to Candida (see Table). Eosinophils and neutrophils were less frequent in Ag+Candida than in Ag sites at 24 hours. These findings indicate that GCR-beta expression is increased at sites of combined Thl and Th2 responses in a combined DIVLPR reaction. Furthermore, it suggests that a relative GC resistance may occur in inflammatory reactions that have the combination of Thl and Th2 responses.
of
Reduced Sensitivity to Dexamethasone (DEX) of Staphylococcal Enterotoxin (SE) B Stimulated Peripheral Blond Mononuclear Cells (PBMC) From Patients With Atopic Dermatitis (AD) PJ Hauk. I Strickland. DYM Leung National Jewish Medical and Research Center, Denver, CO Colonization of the skin by superantigen (SAg) producing microbes has been found to be associated with increased disease severity of AD. These data suggested that stimulation of T cells with superantigens (SAg) may lead to the development of insensitivity to glucocorticoids (GC). To study this possibility more carefully, we assessed the capacity of IO-TM dexamethasone (DEX) vs lpg/ml cyclosporin (Cy) A and IOng/ml FK506 to inhibit the T cell proliferation of PBMC from 7 AD patients and 7 normal controls with the prototypic SAg, SEB, and the mitogen, PHA. In both study groups. DEX caused a 99% inhibition of PHA-induced PBMC proliferation. When compared with DEX, PHA-induced proliferation was significantly less inhibited by CyA in AD patients (60%; p
5267
CELL FREQUENCIES
IN 24 HOURS
BIOPSIES
SITE
GR-BETA+
IL-&
lL5+
AG CAND
3.80.3 3.01.0 11.52.2’ 1SO.8
10.51.3 2.20.3 6.00.9 160.7
12.51.8 2.21.0 570.7** 1.40.5
AG+CAND
BUFFER
‘4’0.01 VS. AG. C SITES, OR BUFFER **-P=O.O03 VS. AG SlTE!S
792
Therapeutic Dermatitis
Potential
SITES
of Specific
Chymase
Teruaki Imada*, Naruyasu Shigeki Kuwahara*, Koji Mizuo Miyazaki j, Tak.oo Kondo*.
Kobayashi*, Yoshikowa*. ra* *Pharmaceutical
Inhibitor Komorito*. Naito*, Norifimi
in Atopic Fuji0 Tsuromu Nakamu-
Research Division, Yoshitomi Pharmaceutical Industries, LTD. Osaka 573-l 153, Japan TDepattment of Pharmacology, Osaka Medical College, Osaka 569-8686. Japan Chymase is a chymotryptic serine protease stored and secreted by mast cells. An increasing body of evidence shows that it possesses diverse effects, including degradation of extracellular matrices, activation of MMPs and direct production of Ang II from angiotensinogen. Its pathophysiological role is however unclear. When purified rat chymase (RMCP-I) was intradetmally injected into mouse, a scratching reflex was observed. In order to investigate the role of chymase in pruritus. we developed orally active chymase-specific inhibitors. One candidate compound, AU0664, showed a strong inhibitory activity against human chymase in vitro (Ki; 22.6 nmol/L). AU0664 ( 3, 10, 30 mg/kg, p.0.) showed dosedependent suppression of the scratching response in a recently established pruritus model using 2,4-dinitrobenzene sulfonate (DNBS)-induced passively sensitized BALB/c mice. It was of interest that AU0664 showed enhanced inhibition of the scratching response in combination with cyproheptadine, a histamine/serotonine antagonist, and predonisolone. This suggests that chymase contributes to the development of pruritus by the mechanism(s) different from those of conventional drugs. In a further experiment, involvement of chymase to the production of IgE was examined in vitro. Purified chymase increased the production of IgE from human and mouse B cells in the presence of IL-4, which was suppressed in the presence of AU0664. This effect was confirmed also in in vivo model. Increased serum IgE level was observed in BN rats injected mercury chloride s.c.. Administration of AU0664 suppressed the response dose-dependently. Finally five repeated topical applications of DNBS to the ears of BALB/c mice resulted in contact dermatitis and elevation of IgE in serum. Orally administered AU0664 lowered ear thickness as well as serum IgE level in a dose-dependent manner. Taken together, these findings suggest that specific chymase inhibitors such as AU0664 have therapeutic potential in the treatment of the symptoms of atopic dermatitis.
J ALLERGY CLIN IMMUNOL VOLUME 104. NUMBER 1, PART 2
studies in these patients as diagnosis of SCID can treatment as well as assist in genetic counseling. 790
791
dictate
priority
were observed at 6 hrs (data not shown). Cells positive for IL-4 or IL-5 mRNA (by in situ hybridization) were less frequent in Ag+Candida sites than in Ag sites possibly due to paracrine suppressive effects on Th2 responses by the Thl responses to Candida (see Table). Eosinophils and neutrophils were less frequent in Ag+Candida than in Ag sites at 24 hours. These findings indicate that GCR-beta expression is increased at sites of combined Thl and Th2 responses in a combined DIVLPR reaction. Furthermore, it suggests that a relative GC resistance may occur in inflammatory reactions that have the combination of Thl and Th2 responses.
of
Reduced Sensitivity to Dexamethasone (DEX) of Staphylococcal Enterotoxin (SE) B Stimulated Peripheral Blond Mononuclear Cells (PBMC) From Patients With Atopic Dermatitis (AD) PJ Hauk. I Strickland. DYM Leung National Jewish Medical and Research Center, Denver, CO Colonization of the skin by superantigen (SAg) producing microbes has been found to be associated with increased disease severity of AD. These data suggested that stimulation of T cells with superantigens (SAg) may lead to the development of insensitivity to glucocorticoids (GC). To study this possibility more carefully, we assessed the capacity of IO-TM dexamethasone (DEX) vs lpg/ml cyclosporin (Cy) A and IOng/ml FK506 to inhibit the T cell proliferation of PBMC from 7 AD patients and 7 normal controls with the prototypic SAg, SEB, and the mitogen, PHA. In both study groups. DEX caused a 99% inhibition of PHA-induced PBMC proliferation. When compared with DEX, PHA-induced proliferation was significantly less inhibited by CyA in AD patients (60%; p
5267
CELL FREQUENCIES
IN 24 HOURS
BIOPSIES
SITE
GR-BETA+
IL-&
lL5+
AG CAND
3.80.3 3.01.0 11.52.2’ 1SO.8
10.51.3 2.20.3 6.00.9 160.7
12.51.8 2.21.0 570.7** 1.40.5
AG+CAND
BUFFER
‘4’0.01 VS. AG. C SITES, OR BUFFER **-P=O.O03 VS. AG SlTE!S
792
Therapeutic Dermatitis
Potential
SITES
of Specific
Chymase
Teruaki Imada*, Naruyasu Shigeki Kuwahara*, Koji Mizuo Miyazaki j, Tak.oo Kondo*.
Kobayashi*, Yoshikowa*. ra* *Pharmaceutical
Inhibitor Komorito*. Naito*, Norifimi
in Atopic Fuji0 Tsuromu Nakamu-
Research Division, Yoshitomi Pharmaceutical Industries, LTD. Osaka 573-l 153, Japan TDepattment of Pharmacology, Osaka Medical College, Osaka 569-8686. Japan Chymase is a chymotryptic serine protease stored and secreted by mast cells. An increasing body of evidence shows that it possesses diverse effects, including degradation of extracellular matrices, activation of MMPs and direct production of Ang II from angiotensinogen. Its pathophysiological role is however unclear. When purified rat chymase (RMCP-I) was intradetmally injected into mouse, a scratching reflex was observed. In order to investigate the role of chymase in pruritus. we developed orally active chymase-specific inhibitors. One candidate compound, AU0664, showed a strong inhibitory activity against human chymase in vitro (Ki; 22.6 nmol/L). AU0664 ( 3, 10, 30 mg/kg, p.0.) showed dosedependent suppression of the scratching response in a recently established pruritus model using 2,4-dinitrobenzene sulfonate (DNBS)-induced passively sensitized BALB/c mice. It was of interest that AU0664 showed enhanced inhibition of the scratching response in combination with cyproheptadine, a histamine/serotonine antagonist, and predonisolone. This suggests that chymase contributes to the development of pruritus by the mechanism(s) different from those of conventional drugs. In a further experiment, involvement of chymase to the production of IgE was examined in vitro. Purified chymase increased the production of IgE from human and mouse B cells in the presence of IL-4, which was suppressed in the presence of AU0664. This effect was confirmed also in in vivo model. Increased serum IgE level was observed in BN rats injected mercury chloride s.c.. Administration of AU0664 suppressed the response dose-dependently. Finally five repeated topical applications of DNBS to the ears of BALB/c mice resulted in contact dermatitis and elevation of IgE in serum. Orally administered AU0664 lowered ear thickness as well as serum IgE level in a dose-dependent manner. Taken together, these findings suggest that specific chymase inhibitors such as AU0664 have therapeutic potential in the treatment of the symptoms of atopic dermatitis.