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116 Mast cells in the epidermis of patients with atopic dermatitis
JSID Abstracts
122
115
118
Genetic analysis of atoplc dermatitis. U. lchimlva.M.~, A.Ohnura, II. Tat.eno. V. Yasui. and C. Anaaam_l Department nf Dermatology Yanaguchl University School of aedlcine,Uhe,Ja~an. in oder to elucidate nusce~llhlIIty to aloplc deraatltla,we lnvestlgaled DNA DOfymOrphfSnS of IILA-DR allele and of mast cell chynanr. Thls study enrolled 51 patients nllh AD.Dlagnosls of AD was based on the criteria proposed by the Japanese Dermatologlcat Assoclatlon The frequency of HLA-DRI and DRl3 allele was signlflcanlly Increased In AD compared vlth those nf normal controls (31.4% VS 10.8% for DRl;D(0.05,20.7% VS 10.8% for DR13:0(0.05). YP conclude I.hat the susceptiblllty gene involved In the development of AD might be in Ilnkage dlsesulllbrla wlth HLA-DRl and/or Drl3. We are examining gcnotyplc 0olymorphlsns of mast cell chynase which night act In the effcctor phase of the disease.
CHARACTERIZATION OF BONE MARROW-DERIVED MAST CJ2LJ.S OBTAINED FROM NC MICE T. w Department of Dermatology, Hiroshima University School of Medicine, Hiroshima, Japan NC mice spontaneously develop atopic cuema-like skin lesion associated with dermal mast cell hyperplasik Skin histamine content of NC was 69.Opg/g wet tissue and significantly higher than those of C57BU6 (16.1&g wet tissue) and BAL5lc (19.4 @g/g wet tissue). To know a mechanism of this mast cell hyperplasia in NC mice, we investigated properties of bone marrow-derived mast cells (BMMC) obtained from NC mice. Histamine content of BMMC from NC was 0.67pg/cell and significantly higher than those of C57BU6 (O.O9pg/cell) and BALBlc (O.O4pg/cell). Percent histamine release in BMMC was found to be similar in these three strains. Adhesion of BMMC to plastic dishes was higher in NC compared with C57BIJ6 and B.&LB/c. These result indicate that NC mlw has an abnormality in histamine content and adhesive pmpelty of mast cells.
116 MAST ATOPIC
119 CELLS IN THE EPIDERMIS DERMATITIS. S.
shib9t@. Departments of Dermatology Kyushu University, Fukuoka, Japan.
OF
and Anatomy,
PATIENTS
WITH
Faculty of Medicine,
We observed mast cells in the epidermis (as well as the dermis) of the skin specimens of atopic dermatitis fixed with Camoy’s, but in none of the conventionally fixed specimens. While mast cells have been identified in the connective tissue of the skin, they have not been observed in the epidermis even in patients with atopic dermatitis. Criteria for identifying mast cells included I) metachromatic response to staining with toluidine blue, 2) positive immunoreaction for anti-human mast cell tryptase monoclonal antibody, and 3) typical ultrastructural features observed on electron microscopy. Observations by immunohistochemistry and electron microscopy confirmed the findings. In all of the specimens, mast cells were seen in the dermal connective tissue, particularly around the capillaries. The fact that special fixation and staining are required to observe mast cells in the epidermis implies that the cells belong to the mucous type. These cells are a source of the cytokines that recruit and activate the inflammatory cells involved in the allergic reaction. Their presence may explain the initiation of the late-phase inflammatory response in the skin of patients with atopic demlatitis.
117 Calcium involvement mast cells H.
FK506 AND CYCLOSPGRIN
A SY INHIBIT STEhf CELL FACTOR-DEPENDENT SIGNAL ‘lRANSDUCI’ION AND UPREGULATE C-KIT BXPRESSION IN MOUSE MAST CELL LINE, MC9 CELLS. . . F. Ito. N.lbvota. H, Department of Dermatology, Asahikawa Malii College, Asahikawa, Japan. Using o~oaaamast cell line, MC/g, we bvmligrtcd lhc effeds of immuooauppnsssnls, SK506 sod cyclosporin A (C&4), on ILor SCFdependentproliferation mddgnd lraud& NdiherPK.506 nor CIA affwkd IG3-
dependentMC/9 cell proliferation, whereaslhsy seladlvdy lahlbiled SC%depaademcell survival. MU9 cells following lheactreatmeats could IIDI be inducedsyoqjistic proliferatbn by both IL3 rndSCF mdmsin(l~edtheirswivd by SCP alone, rendtinS in apoplosir. Flow cytometry showed that lhey dkl not rtisc IL3 recepbx expraaion. However, immunoblol andKGPCR uulyra Micltsd inc?ewd c-kit protein udc-kit mRNA tisluuiptl followlw FICSO6and&A Irclbnenu. In ddbbn, adhaion way mvmkd the bwcaed dhaiw MC/9 cells to NEWI cdkfolbwinS (bae tratment~, mnsklenl with (ha1the bzawed e-kilpmtein is a88wMed with membm&mnd SCP of NlHLW3 cell. The baeaxd c-k@ mc8plor, however, seem to be not functional with mganl to ailpld lmnsduclio~ Neither PK.506nor CdA afle&d c-kil lyrorillc phoapborylation nd MAP kinasa nuckartraodocation following SCF tiimuhtion.
120 in
the
activation
of
cultured
human
O.Ishikawa Department of Dermatology, Gunma University School of Medicine, Maebashi, Japan. Mast cells release various mediators and play an essential role in the development of immediate type allergic reaction. Many investigations of mast cell functions have been conducted using rat peritoneal mast cells. The problem is that mast cells are heterogeneous and there may bc functional differences in mast cells between rats and human beings. Human mononuclear cells were obtained from heparinized umbilical cord blood with Ficoll-paque, and cultured in a medium in the presence of SCF and IL-6. The purity of mast cells reached 100 9%a&r 12 weeks. Mast cells incubated with fura-2/AM were activated, and the fluoresccnsc of fura- was measured by a spectmfluommeter. The fluoresccncc images were also investigated microscopically with an image processor. When IgE-sensitized mast cells were incubated with anti-@, histamine was nleascd in a dose-dependent and time-dependent manner. Calcium ionophore A23187 also induced histamine release. Fluorescence intensity increased obviously in fura-Zloaded mast cells exposed to the stimulators.
INHIBITION OF HISTAMINE RELEASE BY MEDICINAL HERBAL EXTRACTS. A. Hatani’. M. Q&gxhi. H. 0-a. M. ‘l&c& Shiga Central Research Laboratory, NOEVIR Co., Ltd., Shiga, Japan. In general, immediate allergy reaction is one inducer for atopic dermatitis. This allergy is caused by chemical mediators released by basophiles and mast cells on degranulation. Rat baaophilic leukemia cells (RBL2H8 cells) are known to release various chemical mediators induced by DNP-BSA antigen binding to anti-DNP IgE antibodies on the cell surface. We investigated the inhibitory effects of histamine release from RBL-2H3 cells for 200 medicinal herbal extracts. The histamine was measured by post-column HPLC method. The extract of Gambir, Resee fidus, CeryopnUi Flos, Sanguisorba of?icinelIs~ Cinchona@ Cortex, and Betula Ienh inhibited the histamine release from th cells by more than 50%.
122
115
118
Genetic analysis of atoplc dermatitis. U. lchimlva.M.~, A.Ohnura, II. Tat.eno. V. Yasui. and C. Anaaam_l Department nf Dermatology Yanaguchl University School of aedlcine,Uhe,Ja~an. in oder to elucidate nusce~llhlIIty to aloplc deraatltla,we lnvestlgaled DNA DOfymOrphfSnS of IILA-DR allele and of mast cell chynanr. Thls study enrolled 51 patients nllh AD.Dlagnosls of AD was based on the criteria proposed by the Japanese Dermatologlcat Assoclatlon The frequency of HLA-DRI and DRl3 allele was signlflcanlly Increased In AD compared vlth those nf normal controls (31.4% VS 10.8% for DRl;D(0.05,20.7% VS 10.8% for DR13:0(0.05). YP conclude I.hat the susceptiblllty gene involved In the development of AD might be in Ilnkage dlsesulllbrla wlth HLA-DRl and/or Drl3. We are examining gcnotyplc 0olymorphlsns of mast cell chynase which night act In the effcctor phase of the disease.
CHARACTERIZATION OF BONE MARROW-DERIVED MAST CJ2LJ.S OBTAINED FROM NC MICE T. w Department of Dermatology, Hiroshima University School of Medicine, Hiroshima, Japan NC mice spontaneously develop atopic cuema-like skin lesion associated with dermal mast cell hyperplasik Skin histamine content of NC was 69.Opg/g wet tissue and significantly higher than those of C57BU6 (16.1&g wet tissue) and BAL5lc (19.4 @g/g wet tissue). To know a mechanism of this mast cell hyperplasia in NC mice, we investigated properties of bone marrow-derived mast cells (BMMC) obtained from NC mice. Histamine content of BMMC from NC was 0.67pg/cell and significantly higher than those of C57BU6 (O.O9pg/cell) and BALBlc (O.O4pg/cell). Percent histamine release in BMMC was found to be similar in these three strains. Adhesion of BMMC to plastic dishes was higher in NC compared with C57BIJ6 and B.&LB/c. These result indicate that NC mlw has an abnormality in histamine content and adhesive pmpelty of mast cells.
116 MAST ATOPIC
119 CELLS IN THE EPIDERMIS DERMATITIS. S.
shib9t@. Departments of Dermatology Kyushu University, Fukuoka, Japan.
OF
and Anatomy,
PATIENTS
WITH
Faculty of Medicine,
We observed mast cells in the epidermis (as well as the dermis) of the skin specimens of atopic dermatitis fixed with Camoy’s, but in none of the conventionally fixed specimens. While mast cells have been identified in the connective tissue of the skin, they have not been observed in the epidermis even in patients with atopic dermatitis. Criteria for identifying mast cells included I) metachromatic response to staining with toluidine blue, 2) positive immunoreaction for anti-human mast cell tryptase monoclonal antibody, and 3) typical ultrastructural features observed on electron microscopy. Observations by immunohistochemistry and electron microscopy confirmed the findings. In all of the specimens, mast cells were seen in the dermal connective tissue, particularly around the capillaries. The fact that special fixation and staining are required to observe mast cells in the epidermis implies that the cells belong to the mucous type. These cells are a source of the cytokines that recruit and activate the inflammatory cells involved in the allergic reaction. Their presence may explain the initiation of the late-phase inflammatory response in the skin of patients with atopic demlatitis.
117 Calcium involvement mast cells H.
FK506 AND CYCLOSPGRIN
A SY INHIBIT STEhf CELL FACTOR-DEPENDENT SIGNAL ‘lRANSDUCI’ION AND UPREGULATE C-KIT BXPRESSION IN MOUSE MAST CELL LINE, MC9 CELLS. . . F. Ito. N.lbvota. H, Department of Dermatology, Asahikawa Malii College, Asahikawa, Japan. Using o~oaaamast cell line, MC/g, we bvmligrtcd lhc effeds of immuooauppnsssnls, SK506 sod cyclosporin A (C&4), on ILor SCFdependentproliferation mddgnd lraud& NdiherPK.506 nor CIA affwkd IG3-
dependentMC/9 cell proliferation, whereaslhsy seladlvdy lahlbiled SC%depaademcell survival. MU9 cells following lheactreatmeats could IIDI be inducedsyoqjistic proliferatbn by both IL3 rndSCF mdmsin(l~edtheirswivd by SCP alone, rendtinS in apoplosir. Flow cytometry showed that lhey dkl not rtisc IL3 recepbx expraaion. However, immunoblol andKGPCR uulyra Micltsd inc?ewd c-kit protein udc-kit mRNA tisluuiptl followlw FICSO6and&A Irclbnenu. In ddbbn, adhaion way mvmkd the bwcaed dhaiw MC/9 cells to NEWI cdkfolbwinS (bae tratment~, mnsklenl with (ha1the bzawed e-kilpmtein is a88wMed with membm&mnd SCP of NlHLW3 cell. The baeaxd c-k@ mc8plor, however, seem to be not functional with mganl to ailpld lmnsduclio~ Neither PK.506nor CdA afle&d c-kil lyrorillc phoapborylation nd MAP kinasa nuckartraodocation following SCF tiimuhtion.
120 in
the
activation
of
cultured
human
O.Ishikawa Department of Dermatology, Gunma University School of Medicine, Maebashi, Japan. Mast cells release various mediators and play an essential role in the development of immediate type allergic reaction. Many investigations of mast cell functions have been conducted using rat peritoneal mast cells. The problem is that mast cells are heterogeneous and there may bc functional differences in mast cells between rats and human beings. Human mononuclear cells were obtained from heparinized umbilical cord blood with Ficoll-paque, and cultured in a medium in the presence of SCF and IL-6. The purity of mast cells reached 100 9%a&r 12 weeks. Mast cells incubated with fura-2/AM were activated, and the fluoresccnsc of fura- was measured by a spectmfluommeter. The fluoresccncc images were also investigated microscopically with an image processor. When IgE-sensitized mast cells were incubated with anti-@, histamine was nleascd in a dose-dependent and time-dependent manner. Calcium ionophore A23187 also induced histamine release. Fluorescence intensity increased obviously in fura-Zloaded mast cells exposed to the stimulators.
INHIBITION OF HISTAMINE RELEASE BY MEDICINAL HERBAL EXTRACTS. A. Hatani’. M. Q&gxhi. H. 0-a. M. ‘l&c& Shiga Central Research Laboratory, NOEVIR Co., Ltd., Shiga, Japan. In general, immediate allergy reaction is one inducer for atopic dermatitis. This allergy is caused by chemical mediators released by basophiles and mast cells on degranulation. Rat baaophilic leukemia cells (RBL2H8 cells) are known to release various chemical mediators induced by DNP-BSA antigen binding to anti-DNP IgE antibodies on the cell surface. We investigated the inhibitory effects of histamine release from RBL-2H3 cells for 200 medicinal herbal extracts. The histamine was measured by post-column HPLC method. The extract of Gambir, Resee fidus, CeryopnUi Flos, Sanguisorba of?icinelIs~ Cinchona@ Cortex, and Betula Ienh inhibited the histamine release from th cells by more than 50%.