Protein kinase C-δ but not protein kinase C-β is the predominate isoenzyme expressed in...
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22 Oct. 29/Concurrent Session 7 - DermatopathologylElectron Microscopy/Biological Structure 093 LOCALIZATION OF ALKALlNE PHOSPHATASE IN ECCRME AND AFVCRlNE SWEAT GLANDS. ” -Saea Yousuke Morimoto. and Makoto T&ha rhl Depamne”, of Demmtology. Sapporn Medical Unwersity, Sapporo, Japan Alkaline phosphatase (AP) is a group of enzymes dm, are membrane-bound glycoproteins. AP catalyzes the hydrolysis of Inorganic and orgamc monophosphate esters a, alkaline pH. Although this enzyme is widely disoihuled in human tissues. we do”‘, fully understand iu physiological function. The presence of AP 1” the kidney. liver, and intestine has suggested that the enzyme might pamcipae in membrane transpon. The localization of AP in human sweat glands was poorly ondenlcad. Therefore. we med 10 elocida,e the locahzanon of AF’ in human eccrine and apoxine weal glands. Ligh, and elecrro” m,eroscopic enzyme cywchemisuies were employed for the localization of Ap in human sweat glands. Fordemonstranon of APa, the light microscopic level, froze” sections were cot from human skin and fixed in cold acetone. Sections were incubaed I” the reaction medwm that included naphthol AS-MX phosphate and Fast Red RC salt. For demonsuation of Ap a, the electron microscopic level. small pieces of ski” were fixed in the aldehyde fixative. then sweat glands were isola,ed under a stereomicroscope using tweezers. Isolated sweat glands were incubated in the reaction medium that contained P-glycerophosphare and lead ciuate. Thereafter sweat glands were embedded in Epon 812 and thin sections were ohserved.L,ght microscopic enzyme cytochemisuy demonswated strong reaction in the intercellular canaliculi of human eccrine sweat glands, while secretory cells of aparine sweat gland did no, show any rexho”. Myoepithelial cells of eccrine and apocrine sweat glands showed we& reaction. ElecRo” micmscopic enzyme cytochemisuy demonsuated the localization of AP on ihe cell membrane of inlercellulu canabculi of eccrine swea, glands. The luminal cell membrane of eccrine secretory cells that is in continuity with imercellular canaliculi was devoid of lhis reaction. These data suggest tha: the intercellular canalicoli are no, simply a” extension of Ibe luminal cell membrane of eccrine secretory cells. Intercellular canabculi are a differentiated smx,ore tha, may play a role in the prcducnon of primary sweat. 094 AMOUNT OF SECRETORY IMMUNOGLOBuLIN-A DIFFERES WITH SKIN REGION BUT MAINTAINS ITS LEVEL REGARDLESS OF SWEATING. Shrhei Ima m Yuii Shimozmo. Atsumichi U&e_ eeo Ohta. and Keishi Ym, Department of Dermatology @I, AU, YH). Faculty of Medicine, Kyushu University, Fukuoka; Hisamitsu Pharmaceuticals (YS, SO, KY), Tow, Japan We developed a simple method for measuring the amount of the seeretorv form of immunoelobulin A (&A) oresent in sweat. A small disk (1O~lOmrn) made of cellulose man&& was attached to the skin surface for periods of 1 to 24 hours. SIgA was absorbed to the membrane and accumulated during the period of application. An enzyme immunoassay using anti-IgA and anti-secretory component (SC) antibodies revealed dots on the disk which corresponded to the eccrine excretory ducts. A densitograph was used to determine the number and density of the dots, thus obtaining the amount of sIgA excreted to the surface of the skin (per mmz). The amount of skin sIgA excreted in mm*/day by 50 healthy subjects differed inter-individually as well as intra-individuallv: it varied accordine to r&on of the skin. and its distribution roughly’ reflected that of tlhe sweat ducts. SIgA excretion was maintained at a certain level regardless of the increased sweating produced by either heat or exercise, which raised the output of sweat by 3. to 15.fold. Immunohistochemicai studies revealed that fewer glandular cells expressed SC in their cytoplasm as the amount of sIgA decreased. Such an independence of the sIgA excretion from sweat excretion may be necessary to local immune defenses. 095 096 097 BULLOUS CONGENlTAL ICHTHYDSlFORM ERYTHRODERMA OF BROCQ CAUSED BY BOTH DISRUPTWE AND HIGHLY CONSERVATWE AMINO ACID SUBSTITUTIONS IN THE ,A DOMAIN OF KERATINS 1 AND 10. WHIMcLean’.S M Morley’.R A J Ead$.P J C DW’,LB 2 uu&il’,H~Navsarial.C~. uHaK&.DG4._E &Law,. ‘CRC Cell Slructure Research Group. Dep, A”a,omy & Physiology. Universily of Dundee DDl 4HN. UK; zS, John’s l”s,i,u,e of Derma,ology. UMDS. S, Thomas’s Hospilal, London. UK; 3Experime”,al Dermamlogy Laboramries, London Hospital Medical College, London. UK; “Dermalology Deparonc”,. The Hospital for Sick Children, Great Ormond Wee,, land”“. UK. Bullous congenilal ich,hyortform erythroderma (BCIE) is a human au,o~“ma, domina”, ski” disorder characterised by severe epidermalyoc hyperkeralosis Recently, poi”, mutations I” ather of the suprabasal keradna 1 and 10 have bee” show” 1” cause ,his disease The mutations repor,ed 10 da,e are predic,ed ,o be highly disruptive. Here. we rep”” 4 mu,a,lo”s I” Lhe u-helical ,A domain of K, and KlO A pain, mu,a,io” m Kl and another I” KlO produce praline subs,i,ulio”s. which are show” by prmei” s,ruc,ural predic,mnr IO be highly de,nmenta, ID a-helix for”W,o”. I” addition, we report highly conser~~,,~e amin” aad subslilulions in Kl and K10 which also produce ,he d,sorder. ,“,ercS,i”g,y. the phenotype 1s very severe in all 4 pa,ie”,s. indicaring &at even subtle changes in this highly conserved region of kera,i” inlermediate filamenls cannot be ,o,era,ed I” Y~Y” ULTRASTRUCTURAL LOCALIZATION OF BINDING SITES FOR LINEAR IgA BULLOIJS DERMATOSIS (LABD) ANTIBODIES. Cerarv Kowalewrkl. Mnrek Hafrek and Daniel Schmit,. U.346 INSERM / CNRS. Depanme”, ofDemn,ology, Hbpul E. Herrio,, Lyon. France Linear IgA bullous dermatosir (LABD) is charactenmd by the presence of linen IgA deposirs in rhe cu,:u~eoos basement membrane zone A, leas, two patterns of deposi,io” can be deteard by direc, ~mnwoelectro” microscopy (IEM): I” rhe lamina lucIda (LL) and I” ,he sub- lamina densa (LD) zone A parocular “mirror Image” pattern combining deposirs on both sides of ,he LD has bee” also described.. I” ,he prc\c”, ,“d,rect IEM \,udy, sera of 26 pauentr wrh LABD were used for prrclw ~oc~~~~zB,~o” of ,hc ,arge, :~n,ige”s I” IWO substraw: normal human ski” and monkey esophagus. l’hc scr;l were relrc,cd ualng ,“dlrec, m~“,u”ofluorescence, on ,he bss,z of then reac,w,,y wnh \a],-rpl,, skm: 24 sera bound ,o the roof of rhe bulla, I to ,he roof lind ,he floor, and 2 10 rhe floor alone. 0” IEM. ,hc larier two sera gave a predomt”.,“, sub~LD Immulloperoxidasie labeling. however. a concomitant hnear ~,al”l”g I” the LL was ialso observed Out of rhe rema,“,“g 24 sera. 13 sralned hem,desmo\omer (50%). Y labeled the LL t34.6%), and two were u”rex”ve (on enher substrate). Most sew reacted w,rh monkey esophagus, whereas several ones were “egarlve on human rkl” For these reactmg wrh both ,issues. the labelmg pattern was alway\ reproduublc A LABD sew”, recognnng the 97 kD pro,el” (I J Zone et al.. J Cl,” ,“ves, XS X12-X20,lYYO) labeled the LL. Control sera contalmng IgG autoanubodies agalnr, ,he 180 kD and 230 kD bullous pemphigoid and the 290 kD epldermolysn bulloaa acqu~uta anrlgena reac,ed wth the LL. hemidesmosomes, and wb-LD zone. respecwely No relauomhip between the clinical p,c,ure and the obselved \,;immg pairem of LABD sera could be found. Our rewltr conRam ,he hererogeneous “ilure of LABD bu, do no, support ,he concept of ch”,c:~I s~g”~f,cancc of any px,icular lofahzatio” of binding bites for LABD autoanrihodn wthin the basemen, memhmne zone.
Transcript of Protein kinase C-δ but not protein kinase C-β is the predominate isoenzyme expressed in...
22 Oct. 29/Concurrent Session 7 - DermatopathologylElectron Microscopy/Biological Structure
093 LOCALIZATION OF ALKALlNE PHOSPHATASE IN ECCRME AND AFVCRlNE SWEAT GLANDS. ” -Saea Yousuke Morimoto. and Makoto T&ha rhl Depamne”, of Demmtology. Sapporn Medical Unwersity, Sapporo, Japan
Alkaline phosphatase (AP) is a group of enzymes dm, are membrane-bound glycoproteins. AP catalyzes the hydrolysis of Inorganic and orgamc monophosphate esters a, alkaline pH. Although this enzyme is widely disoihuled in human tissues. we do”‘, fully understand iu physiological function. The presence of AP 1” the kidney. liver, and intestine has suggested that the enzyme might pamcipae in membrane transpon. The localization of AP in human sweat glands was poorly ondenlcad. Therefore. we med 10 elocida,e the locahzanon of AF’ in human eccrine and apoxine weal glands. Ligh, and elecrro” m,eroscopic enzyme cywchemisuies were employed for the localization of Ap in human sweat glands. Fordemonstranon of APa, the light microscopic level, froze” sections were cot from human skin and fixed in cold acetone. Sections were incubaed I” the reaction medwm that included naphthol AS-MX phosphate and Fast Red RC salt. For demonsuation of Ap a, the electron microscopic level. small pieces of ski” were fixed in the aldehyde fixative. then sweat glands were isola,ed under a stereomicroscope using tweezers. Isolated sweat glands were incubated in the reaction
medium that contained P-glycerophosphare and lead ciuate. Thereafter sweat glands were
embedded in Epon 812 and thin sections were ohserved.L,ght microscopic enzyme cytochemisuy demonswated strong reaction in the intercellular canaliculi of human eccrine sweat glands, while secretory cells of aparine sweat gland did no, show any rexho”. Myoepithelial cells of eccrine and apocrine sweat glands showed we& reaction. ElecRo” micmscopic enzyme cytochemisuy demonsuated the localization of AP on ihe cell membrane of inlercellulu canabculi of eccrine swea, glands. The luminal cell membrane of eccrine secretory cells that is in continuity with imercellular canaliculi was devoid of lhis reaction. These data suggest tha: the intercellular canalicoli are no, simply a” extension of Ibe luminal cell membrane of eccrine secretory cells. Intercellular canabculi are a differentiated smx,ore tha, may play a role in the prcducnon of primary sweat.
094
AMOUNT OF SECRETORY IMMUNOGLOBuLIN-A DIFFERES WITH
SKIN REGION BUT MAINTAINS ITS LEVEL REGARDLESS OF SWEATING. Shrhei Ima m Yuii Shimozmo. Atsumichi U&e_
eeo Ohta. and Keishi Ym, Department of Dermatology @I, AU, YH). Faculty of Medicine, Kyushu University, Fukuoka; Hisamitsu Pharmaceuticals (YS, SO, KY), Tow, Japan
We developed a simple method for measuring the amount of the seeretorv form of immunoelobulin A (&A) oresent in sweat. A small disk (1O~lOmrn) made of cellulose man&& was attached to the skin surface for periods of 1 to 24 hours. SIgA was absorbed to the membrane and accumulated during the period of application. An enzyme immunoassay using anti-IgA and anti-secretory component (SC) antibodies revealed dots on the disk which corresponded to the eccrine excretory ducts. A densitograph was used to determine the number and density of the dots, thus obtaining the amount of sIgA excreted to the surface of the skin (per mmz). The amount of skin sIgA excreted in mm*/day by 50 healthy subjects differed inter-individually as well as intra-individuallv: it varied accordine to r&on of the skin. and its distribution roughly’ reflected that of tlhe sweat ducts. SIgA excretion was maintained at a certain level regardless of the increased sweating produced by either heat or exercise, which raised the output of sweat by 3. to 15.fold. Immunohistochemicai studies revealed that fewer glandular cells expressed SC in their cytoplasm as the amount of sIgA decreased. Such an independence of the sIgA excretion from sweat excretion may be necessary to local immune defenses.
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097 BULLOUS CONGENlTAL ICHTHYDSlFORM ERYTHRODERMA OF BROCQ CAUSED BY BOTH DISRUPTWE AND HIGHLY CONSERVATWE AMINO ACID SUBSTITUTIONS IN THE ,A DOMAIN OF KERATINS 1 AND 10.
WHIMcLean’.S M Morley’.R A J Ead$.P J C DW’,LB
2 uu&il’,H~Navsarial.C~. uHaK&.DG4._E
&Law,. ‘CRC Cell Slructure Research Group. Dep, A”a,omy & Physiology.
Universily of Dundee DDl 4HN. UK; zS, John’s l”s,i,u,e of Derma,ology.
UMDS. S, Thomas’s Hospilal, London. UK; 3Experime”,al Dermamlogy
Laboramries, London Hospital Medical College, London. UK; “Dermalology Deparonc”,. The Hospital for Sick Children, Great Ormond Wee,, land”“. UK.
Bullous congenilal ich,hyortform erythroderma (BCIE) is a human au,o~“ma, domina”, ski” disorder characterised by severe epidermalyoc hyperkeralosis Recently, poi”, mutations I” ather of the suprabasal keradna 1 and 10 have bee” show” 1” cause ,his disease The mutations repor,ed 10 da,e are predic,ed ,o be highly disruptive. Here. we rep”” 4 mu,a,lo”s I” Lhe u-helical ,A domain of K, and KlO A pain, mu,a,io” m Kl and another I” KlO produce praline subs,i,ulio”s. which are show” by prmei” s,ruc,ural predic,mnr IO be highly de,nmenta, ID a-helix for”W,o”. I” addition, we report highly conser~~,,~e amin” aad subslilulions in Kl and K10 which also produce ,he d,sorder. ,“,ercS,i”g,y. the phenotype 1s very severe in all 4 pa,ie”,s. indicaring &at even subtle changes in this highly conserved region of kera,i” inlermediate filamenls cannot be ,o,era,ed I” Y~Y”
ULTRASTRUCTURAL LOCALIZATION OF BINDING SITES FOR LINEAR IgA BULLOIJS DERMATOSIS (LABD) ANTIBODIES. Cerarv Kowalewrkl. Mnrek Hafrek and Daniel Schmit,. U.346 INSERM / CNRS. Depanme”, ofDemn,ology, Hbpul E. Herrio,, Lyon. France
Linear IgA bullous dermatosir (LABD) is charactenmd by the presence of linen IgA deposirs in rhe cu,:u~eoos basement membrane zone A, leas, two patterns of deposi,io” can be deteard by direc, ~mnwoelectro” microscopy (IEM): I” rhe lamina lucIda (LL) and I” ,he sub- lamina densa (LD) zone A parocular “mirror Image” pattern combining deposirs on both sides of ,he LD has bee” also described..
I” ,he prc\c”, ,“d,rect IEM \,udy, sera of 26 pauentr wrh LABD were used for prrclw ~oc~~~~zB,~o” of ,hc ,arge, :~n,ige”s I” IWO substraw: normal human ski” and monkey esophagus. l’hc scr;l were relrc,cd ualng ,“dlrec, m~“,u”ofluorescence, on ,he bss,z of then reac,w,,y wnh \a],-rpl,, skm: 24 sera bound ,o the roof of rhe bulla, I to ,he roof lind ,he floor, and 2 10 rhe floor alone. 0” IEM. ,hc larier two sera gave a predomt”.,“, sub~LD Immulloperoxidasie labeling. however. a concomitant hnear ~,al”l”g I” the LL was ialso observed Out of rhe rema,“,“g 24 sera. 13 sralned hem,desmo\omer (50%). Y labeled the LL t34.6%), and two were u”rex”ve (on enher substrate). Most sew reacted w,rh monkey esophagus, whereas several ones were “egarlve on human rkl” For these reactmg wrh both ,issues. the labelmg pattern was alway\ reproduublc A LABD sew”, recognnng the 97 kD pro,el” (I J Zone et al.. J Cl,” ,“ves, XS X12-X20,lYYO) labeled the LL. Control sera contalmng IgG autoanubodies agalnr, ,he 180 kD and 230 kD bullous pemphigoid and the 290 kD epldermolysn bulloaa acqu~uta anrlgena reac,ed wth the LL. hemidesmosomes, and wb-LD zone. respecwely No relauomhip between the clinical p,c,ure and the obselved \,;immg pairem of LABD sera could be found.
Our rewltr conRam ,he hererogeneous “ilure of LABD bu, do no, support ,he concept of ch”,c:~I s~g”~f,cancc of any px,icular lofahzatio” of binding bites for LABD autoanrihodn wthin the basemen, memhmne zone.
