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IGE antibodies in sera from patients with bullous pemphigoid are autoantibodies exclusively directed against the 230kDa epidermal antigen (BP230)
ESDR I JSID I SID Abstracts
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0247 CHARACTBRIZATION OF BULLOUS PEMPHIGOID ANTIBODY IN CLINICALLY ACTIVE AND REMISSION STAGE. Masavuki Suzukil, Shieenori Harada? S&n&d -1, -1 and Hidemi -1, LDeptiment of Dermatology, Jichi Medical School, Tochigi Z.Deputy General Manager Kuise Plant Support Shionogi Co. Hyogo, Japan. To examine the quality differences of Bullous pemphigoid (BP) IgG, we purified IgG from BP patients sera in clinically active and remission stage, and performed the examination, as follows. Firstly, using indirect immunofluorescence. we examined antibasement membrane zone (BMZ) antibody activity of IgG subclass. Secondly using enzyme-linked immunosorbent assay(ELISA), we examined binding capability of BP IgG subclass against NC 16a, Pl-2 and Pl-1. Thirdly using flue3, we examined the change of [C&t ]i in keratinccytes, which was induced by BP IgG. Firstly IgGl and IgG4 were the main IgG subclass of BP antibody. The changes of anti-Bh4Z IgG subclass titers were considerably variable among patients. The binding capability to Ncl6a approximately correlated with anti-BMZ IgG subclass titer. The binding capability of IgGI and IgG4 from one of three BP patients to Pl-2 and PI-l decreased in remission stage. Thirdly a fluorescent intensity increase was observed in three of 6 BP IgG of active stage, but not in the same BP IgG of remission stage. These BP IgG of remission stage did not block the fluorescent intensity increase of flue-3 induced by BP IgG of active stage. These results demonstrated that there was no quality differences of BP IgG from active and remission stage.
0251
0248 DETERMINATION OF SUSCEPTIBILITY FACTORS IN BULLQUS PEMPIIIGOID: GENETIC POLYMORPHISMS OF IMMUNOGLOBLJLIN LIGHT-CHAIN GENES. G. Rnur*, D. Gilbert*, P. Joly”, M. Daveau+, Ph. Lauret and F. Tron*. “GRIMP, HBpital Charles Nicolle, Rouen; +INSERM U78, Bois-Guillaume, Institut
F&dCratif de Rechercht Multidisciplinsire les peptidu @X23), France.
mr
Bullous pemphigoid (BP) is an autoimmune blistering skin disease characterized by the production of auto&bodies directed against the basement membrane zone of stratified squamous epithelium. BP antigens consist of two major antigens with molecular weights of 230~kDa (BPAGl) and 180~kDa (BPAGZ). The anti-BMZ antibodies of patients showed an isotype restriction to IgGl and lgG4 subclasses and B predominance of Y light chains. To investigate wether the immunaglohuiin light chain genes influence the occwence of the disease we determined allotypic markers of x chains (Km) using a method combining specific amplification of the constant segment Ck by PCR and subsequent restriction enzyme digestion. A group of 80 patients was compared to 240 healthy individuals ethnically matched to the disease group. We found a positive association between the Km (1, 2) allotype and susceptibility to bullous pemphigoid.
IGE ANTIBODIES IN SERA FROM PATIENTS WITH BIJLLOUS PEMPHIGOID
Dermatology.I?.Heniot Hospital,ClaudeBernardUniversity.Lyon, France. Bullous cemvhieoid (BP) is uniaue amoneautoimmuneskindiseases in which a high wum IgE level
liavebee~ d&ted.
A NOVEL EPITOPE WITHIN THE BP180 NCl6A DOMAIN IS TARGETED BY AUTOANTIBODIES IN LICHEN PLANUS PEMPHIGOIDES. D. Zillikens’.2. F. Cauti’. J.M. Mascaro’. C Prost’. H. Krenia’. E SchmidT
.P. Callen’, E.B. Brilcker’. L.A. Dial’. and G.J. Giudice.“’ Depts Dermatol, ‘Umv Wllrzburg, Germany, ‘Med Coil Wisconsin, Milwaukee, WI, ‘Hospital St. Luis, Paris, France, ‘Univ Louisville, KY, and ‘Dept Biochem. Med Coil Wisconsin, Milwaukee, WI, USA.. Lichen planus pemphigoides (LPP) is an autoimmune subepidermal blistering disease, The finding of IgG antibodies directed against the basement membrane zone differentiates it from bullous LP. The aim of the present study was to determine whether LPP sera react with BP160, a hemidesmosomal protein that is known to be targeted by bullous pemphigoid (BP) autoantibodies. The four LPP sera used in this study stained the epidermal side of NaCI-split human skin in a fashion similar to that obtained with BP sera. By immunoblotting and ELISA, all 4 LPP sera were strongly reactive with the BP160 NC16A domain, the non-collagenous stretch known to contain the major BP autoantibody-reactive sites. The LPP sera showed the highest reactivity with a 14 amino acid stretch of NCIGA that was previously shown to be unreactive with BP sera. Two LPP sera, in addition, reacted with the immunodominant antiganic region associated with BP. In conclusion, there are now four bullous diseases BP, herpes gestationis, cicatricial pemphigoid, and LPP -- that are associated with an autoimmune response against BP180. A novel epitope within the BP180 NClGA domain, designated MCW-4, appears to be uniquely recognized by LPP sera.
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with either an epidermal componentof basementmembrane zone by immunofluorescence microscopy on 1 M NaCt-split tin 01 with a 23OkDa anugen by immunoblots of cultured human keratinocytes. The 230kDa epidennal antigen recognized by IgE antibodies. co-
migrated with the BP230 as labeled by a specific human monoclonal antibody. IgE antiBP230 antibodies in patients’sera were always associatedwith the IgG autoantibodies.ND sera contained IgE antibodies to the 180kDa.BP180or to any other epidermal or dermal antigens BS verified by immunoblot and a newly developed ELISA using the most immunogenic part of BP180. A good correlation was found between the presence of IgE circulatingautoantitadies and the level of serumIgB (P
0249
.
We soughtto determinethe anti~etic wcificity of theseIgE antibodiesin 39 BP sem by immuriofluorescencemicmw6py. itimunobiot and ELiSA. Tbe patient’s sera contained IgG ant&dies to 230kDa(BP230)(n=20).1XOkDa (BP180)(n=9)and to the both BP230and BP180 (n=lO).Senxn IgE levels varied fmm 29 to 5oM)KIIJ/l (meaniSD: 856f1426) among which sem containing IgG antibodiesto BP230 had an IgE level in average 4.3 times more than anti-BP180sem..I@ antiibodiesin eighteensera werefound to be autoantibodiesreactive
0252
0250
0247 CHARACTBRIZATION OF BULLOUS PEMPHIGOID ANTIBODY IN CLINICALLY ACTIVE AND REMISSION STAGE. Masavuki Suzukil, Shieenori Harada? S&n&d -1, -1 and Hidemi -1, LDeptiment of Dermatology, Jichi Medical School, Tochigi Z.Deputy General Manager Kuise Plant Support Shionogi Co. Hyogo, Japan. To examine the quality differences of Bullous pemphigoid (BP) IgG, we purified IgG from BP patients sera in clinically active and remission stage, and performed the examination, as follows. Firstly, using indirect immunofluorescence. we examined antibasement membrane zone (BMZ) antibody activity of IgG subclass. Secondly using enzyme-linked immunosorbent assay(ELISA), we examined binding capability of BP IgG subclass against NC 16a, Pl-2 and Pl-1. Thirdly using flue3, we examined the change of [C&t ]i in keratinccytes, which was induced by BP IgG. Firstly IgGl and IgG4 were the main IgG subclass of BP antibody. The changes of anti-Bh4Z IgG subclass titers were considerably variable among patients. The binding capability to Ncl6a approximately correlated with anti-BMZ IgG subclass titer. The binding capability of IgGI and IgG4 from one of three BP patients to Pl-2 and PI-l decreased in remission stage. Thirdly a fluorescent intensity increase was observed in three of 6 BP IgG of active stage, but not in the same BP IgG of remission stage. These BP IgG of remission stage did not block the fluorescent intensity increase of flue-3 induced by BP IgG of active stage. These results demonstrated that there was no quality differences of BP IgG from active and remission stage.
0251
0248 DETERMINATION OF SUSCEPTIBILITY FACTORS IN BULLQUS PEMPIIIGOID: GENETIC POLYMORPHISMS OF IMMUNOGLOBLJLIN LIGHT-CHAIN GENES. G. Rnur*, D. Gilbert*, P. Joly”, M. Daveau+, Ph. Lauret and F. Tron*. “GRIMP, HBpital Charles Nicolle, Rouen; +INSERM U78, Bois-Guillaume, Institut
F&dCratif de Rechercht Multidisciplinsire les peptidu @X23), France.
mr
Bullous pemphigoid (BP) is an autoimmune blistering skin disease characterized by the production of auto&bodies directed against the basement membrane zone of stratified squamous epithelium. BP antigens consist of two major antigens with molecular weights of 230~kDa (BPAGl) and 180~kDa (BPAGZ). The anti-BMZ antibodies of patients showed an isotype restriction to IgGl and lgG4 subclasses and B predominance of Y light chains. To investigate wether the immunaglohuiin light chain genes influence the occwence of the disease we determined allotypic markers of x chains (Km) using a method combining specific amplification of the constant segment Ck by PCR and subsequent restriction enzyme digestion. A group of 80 patients was compared to 240 healthy individuals ethnically matched to the disease group. We found a positive association between the Km (1, 2) allotype and susceptibility to bullous pemphigoid.
IGE ANTIBODIES IN SERA FROM PATIENTS WITH BIJLLOUS PEMPHIGOID
Dermatology.I?.Heniot Hospital,ClaudeBernardUniversity.Lyon, France. Bullous cemvhieoid (BP) is uniaue amoneautoimmuneskindiseases in which a high wum IgE level
liavebee~ d&ted.
A NOVEL EPITOPE WITHIN THE BP180 NCl6A DOMAIN IS TARGETED BY AUTOANTIBODIES IN LICHEN PLANUS PEMPHIGOIDES. D. Zillikens’.2. F. Cauti’. J.M. Mascaro’. C Prost’. H. Krenia’. E SchmidT
.P. Callen’, E.B. Brilcker’. L.A. Dial’. and G.J. Giudice.“’ Depts Dermatol, ‘Umv Wllrzburg, Germany, ‘Med Coil Wisconsin, Milwaukee, WI, ‘Hospital St. Luis, Paris, France, ‘Univ Louisville, KY, and ‘Dept Biochem. Med Coil Wisconsin, Milwaukee, WI, USA.. Lichen planus pemphigoides (LPP) is an autoimmune subepidermal blistering disease, The finding of IgG antibodies directed against the basement membrane zone differentiates it from bullous LP. The aim of the present study was to determine whether LPP sera react with BP160, a hemidesmosomal protein that is known to be targeted by bullous pemphigoid (BP) autoantibodies. The four LPP sera used in this study stained the epidermal side of NaCI-split human skin in a fashion similar to that obtained with BP sera. By immunoblotting and ELISA, all 4 LPP sera were strongly reactive with the BP160 NC16A domain, the non-collagenous stretch known to contain the major BP autoantibody-reactive sites. The LPP sera showed the highest reactivity with a 14 amino acid stretch of NCIGA that was previously shown to be unreactive with BP sera. Two LPP sera, in addition, reacted with the immunodominant antiganic region associated with BP. In conclusion, there are now four bullous diseases BP, herpes gestationis, cicatricial pemphigoid, and LPP -- that are associated with an autoimmune response against BP180. A novel epitope within the BP180 NClGA domain, designated MCW-4, appears to be uniquely recognized by LPP sera.
-
with either an epidermal componentof basementmembrane zone by immunofluorescence microscopy on 1 M NaCt-split tin 01 with a 23OkDa anugen by immunoblots of cultured human keratinocytes. The 230kDa epidennal antigen recognized by IgE antibodies. co-
migrated with the BP230 as labeled by a specific human monoclonal antibody. IgE antiBP230 antibodies in patients’sera were always associatedwith the IgG autoantibodies.ND sera contained IgE antibodies to the 180kDa.BP180or to any other epidermal or dermal antigens BS verified by immunoblot and a newly developed ELISA using the most immunogenic part of BP180. A good correlation was found between the presence of IgE circulatingautoantitadies and the level of serumIgB (P
0249
.
We soughtto determinethe anti~etic wcificity of theseIgE antibodiesin 39 BP sem by immuriofluorescencemicmw6py. itimunobiot and ELiSA. Tbe patient’s sera contained IgG ant&dies to 230kDa(BP230)(n=20).1XOkDa (BP180)(n=9)and to the both BP230and BP180 (n=lO).Senxn IgE levels varied fmm 29 to 5oM)KIIJ/l (meaniSD: 856f1426) among which sem containing IgG antibodiesto BP230 had an IgE level in average 4.3 times more than anti-BP180sem..I@ antiibodiesin eighteensera werefound to be autoantibodiesreactive
0252