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Autoantibodies to epiligrin are a specific marker for a unique form of cicatricial pemphigoid (CP)
Oct. SO/Concurrent
Session 12 - Bdous
Diseases
172
169 IDENTIFICATION OF A BASEMENT MEMBRANE ZONE ANTIGEN REACTIVE WITH A CIRCULATING IgA ANTIBODY IN OCULAR CICATRICIAL PEMPHIGOID. FP Smith. TB Tr&t_(.J Mew d JJ Zo “t& Division of Dermatology, University of Utah, Sait Lake City, U:. L&A Ocular dcatricial pemphigoid (OCP) is characterized by linear deposition of IgG and/or IgA along the basement membrane zone (BMZ) in conjunctival biopsies, but the target antigen responsible for the immune response has never been identified. We evaluated the antiaenic soecificitv of OCP sera using Western blots against epidermal extra& and &wndaffinity purified antibodies. Our patient selection was based on the presence of clinical evidence of ocular disease (excluding those with oral and skin disease) and a positive direct immunofluorescence (IF) of conjunctival biopsies for IgG and/or IgA. Indirect IF revealed only IgA (not IgG) antibodies. lmmunobloning wth epidenal extracts was performed using sera from OCP patients and an CCP antibody directed against a 45 kD_antigen was found in all seven patients enrolled in our study. Western blots and separated skin were used as immunoafiinity substrates for purtfication of an IgA antigen-specific antibody. This immunoaffinity purrfied antibody bound to the 45 kD region on Western blot and bound in a linear fashion to the BMZ on separated skin. This IoA antibodv failed to show soecificitv to the 230 or 180 kD bullous pemprhigoid antigens and the 97 kD linea; IgA bullous dermatosis antigen. We conclude that OCP sera contain a unique IgA antibody that binds to a 45 kD BMZ antigen.
a
le have obtained 9 sera which clexly dewnstrated noncurrent anti-intercellular (10 and anti-basement membranezone IBYZ) IgC autoantibodies with i-unofluorescenee of buan skin section. These cases are clinicallr diagnosed as buIIous wmpbigoid (BP), gestationis, BP vegetans or -bigus foliaceus (PF). The purpose of the present study was to identify the antigens recognized by these sera with i-unoblot analysis using nonal humanwide-l extracts Iseparated by either EDT* or diswse treatment) and bovine muzzle de-preparation. A numberof possible antigen moleculeswere detected by these sera in various patterns; i.e.. desmoplakins IDPI I/n. desmoglein (PF antipen), Paphigus vulgaris antigen. dewcollins I/II. the 230 kD BP antigen and the 180 bD BP antigen wre detected by 3. 4, I. 1. 6. and 4 aura. respectiWAY. One serum frrw BP-Like case reactive with DP VII clearly stained the intercalate disks sitb ivunafluorescenee of humanand mousecardiac muscle section. suggesting the anti-DP antibodies are responsible to its IC staining in the epidermis. These results indicate that the cases with concurrentanti_IC and anti-M2 antibodies are bigblr heterogeneousand that sac of tba SW rewesent the eoexsistence or intermediate trpe of BP and papbiws.
herpes
170 Eprtope
mapping
sera by moleculariy Toshlhiro Tanaka,
of
Eprdermolysis
Bullosa
Aquisita
patient
cloned cDNA for type VII collagen. Fukuml Furukawa. and Sadao lmamura
Department of Dermatology, Faculty of Medrcrne. Kyoto Umversrty, Kyoto 606, Japan. Eptdermolysis bullosa acquwta IS a subeptdermal blistermg disease rn whtch pattents have autoantibodtes agamst the nonStarting with collagenous domarn of type VII collagen previously Isolated IKb overlapping cDNAs wtth -3 2Kb, encodmg most
cDNA for thus autoantlgen, we Isolated a combined open reading frame of (-115KDa) of the N-terminal nonTo localize collagenous domarn of type VII collagen immunogenic domain, we produced the maltose brnding protein fusion protem with cDNA encodmg different portrons of thts lmmunoblot analysrs of these fusion protein wtth autoantlgen. eprdermolysrs bullosa acqurstta that each patient serum bmds molecule throughout epitopes collagen patrents
pattent sera demonstrated to drfferent region of this
and that epttopes for these patient sera locate These data suggest that multiple thus autoanttgen on the N-terminal non-collagenous domain of type VII are recogntzed by crrculating autoantrbodies m wth eprdermolysis bullosa acqulslta.
174
171 AlJTOANTIBODIES UNIQUE FORM
z.
hzarova.
TO EPILlGRIN OF CICATRICIAL G.J. mhalt. c. Huff. and N.
RRE
A
SPECIFIC
PEMPHIGCJID W.R. Gammon. Daml”~e-“ultsch.
[CP) R.R.
MARKER FOR A K.B. Yance”, Briscmman,
Bethesda, MD, Hill, NC, Washington, DC, Denver, CO. Baltimore. MD, chapel The malor integrin liqand in cultured human keratinocyte (HK) extracellular matrix (ECM) is a glycoprotein complex epiliqrin 1s found in the In human skin, called eplliqrln. lamina lucida sabreqian of epidermal basement membrane (BM) where it is thought to be associated with anchoring We have identified six patients who have IqG filaments. autoantibodies that stain HK KM, bind the superior aspect of lamina densa in 1 M Nacl split skin, and immunoprecipitate (IP) a set of disulfide-linked polypeptides in and ECM in the same manner as cell extracts, HK media PlEl anti-epilrqrin monoclonal antibody (ilAb) and MAb GB3. Studies of other patients with the same clinical phenotype found that while seven have anti-EM (i.e., autoantibodies (IqG, N=S:IqA, N=x), these autoantibodies are directed exclusively against the epidermal side of present In low titer (i.e., < 1:12), 1 M NaCl split skin, nonreactive with HK ECM, and negative in IP studies of HK These findings indicate that CP media and cell extracts. is a disease phenotype in which patlent autoantibodies tarclet different BM constituents and that anti-epillgrin autoantibodies are a specific marker for a unique form of Cp that we propose to call anti-epiliqrin Cp.
M.
Welch.
CP)(N=10)
Session 12 - Bdous
Diseases
172
169 IDENTIFICATION OF A BASEMENT MEMBRANE ZONE ANTIGEN REACTIVE WITH A CIRCULATING IgA ANTIBODY IN OCULAR CICATRICIAL PEMPHIGOID. FP Smith. TB Tr&t_(.J Mew d JJ Zo “t& Division of Dermatology, University of Utah, Sait Lake City, U:. L&A Ocular dcatricial pemphigoid (OCP) is characterized by linear deposition of IgG and/or IgA along the basement membrane zone (BMZ) in conjunctival biopsies, but the target antigen responsible for the immune response has never been identified. We evaluated the antiaenic soecificitv of OCP sera using Western blots against epidermal extra& and &wndaffinity purified antibodies. Our patient selection was based on the presence of clinical evidence of ocular disease (excluding those with oral and skin disease) and a positive direct immunofluorescence (IF) of conjunctival biopsies for IgG and/or IgA. Indirect IF revealed only IgA (not IgG) antibodies. lmmunobloning wth epidenal extracts was performed using sera from OCP patients and an CCP antibody directed against a 45 kD_antigen was found in all seven patients enrolled in our study. Western blots and separated skin were used as immunoafiinity substrates for purtfication of an IgA antigen-specific antibody. This immunoaffinity purrfied antibody bound to the 45 kD region on Western blot and bound in a linear fashion to the BMZ on separated skin. This IoA antibodv failed to show soecificitv to the 230 or 180 kD bullous pemprhigoid antigens and the 97 kD linea; IgA bullous dermatosis antigen. We conclude that OCP sera contain a unique IgA antibody that binds to a 45 kD BMZ antigen.
a
le have obtained 9 sera which clexly dewnstrated noncurrent anti-intercellular (10 and anti-basement membranezone IBYZ) IgC autoantibodies with i-unofluorescenee of buan skin section. These cases are clinicallr diagnosed as buIIous wmpbigoid (BP), gestationis, BP vegetans or -bigus foliaceus (PF). The purpose of the present study was to identify the antigens recognized by these sera with i-unoblot analysis using nonal humanwide-l extracts Iseparated by either EDT* or diswse treatment) and bovine muzzle de-preparation. A numberof possible antigen moleculeswere detected by these sera in various patterns; i.e.. desmoplakins IDPI I/n. desmoglein (PF antipen), Paphigus vulgaris antigen. dewcollins I/II. the 230 kD BP antigen and the 180 bD BP antigen wre detected by 3. 4, I. 1. 6. and 4 aura. respectiWAY. One serum frrw BP-Like case reactive with DP VII clearly stained the intercalate disks sitb ivunafluorescenee of humanand mousecardiac muscle section. suggesting the anti-DP antibodies are responsible to its IC staining in the epidermis. These results indicate that the cases with concurrentanti_IC and anti-M2 antibodies are bigblr heterogeneousand that sac of tba SW rewesent the eoexsistence or intermediate trpe of BP and papbiws.
herpes
170 Eprtope
mapping
sera by moleculariy Toshlhiro Tanaka,
of
Eprdermolysis
Bullosa
Aquisita
patient
cloned cDNA for type VII collagen. Fukuml Furukawa. and Sadao lmamura
Department of Dermatology, Faculty of Medrcrne. Kyoto Umversrty, Kyoto 606, Japan. Eptdermolysis bullosa acquwta IS a subeptdermal blistermg disease rn whtch pattents have autoantibodtes agamst the nonStarting with collagenous domarn of type VII collagen previously Isolated IKb overlapping cDNAs wtth -3 2Kb, encodmg most
cDNA for thus autoantlgen, we Isolated a combined open reading frame of (-115KDa) of the N-terminal nonTo localize collagenous domarn of type VII collagen immunogenic domain, we produced the maltose brnding protein fusion protem with cDNA encodmg different portrons of thts lmmunoblot analysrs of these fusion protein wtth autoantlgen. eprdermolysrs bullosa acqurstta that each patient serum bmds molecule throughout epitopes collagen patrents
pattent sera demonstrated to drfferent region of this
and that epttopes for these patient sera locate These data suggest that multiple thus autoanttgen on the N-terminal non-collagenous domain of type VII are recogntzed by crrculating autoantrbodies m wth eprdermolysis bullosa acqulslta.
174
171 AlJTOANTIBODIES UNIQUE FORM
z.
hzarova.
TO EPILlGRIN OF CICATRICIAL G.J. mhalt. c. Huff. and N.
RRE
A
SPECIFIC
PEMPHIGCJID W.R. Gammon. Daml”~e-“ultsch.
[CP) R.R.
MARKER FOR A K.B. Yance”, Briscmman,
Bethesda, MD, Hill, NC, Washington, DC, Denver, CO. Baltimore. MD, chapel The malor integrin liqand in cultured human keratinocyte (HK) extracellular matrix (ECM) is a glycoprotein complex epiliqrin 1s found in the In human skin, called eplliqrln. lamina lucida sabreqian of epidermal basement membrane (BM) where it is thought to be associated with anchoring We have identified six patients who have IqG filaments. autoantibodies that stain HK KM, bind the superior aspect of lamina densa in 1 M Nacl split skin, and immunoprecipitate (IP) a set of disulfide-linked polypeptides in and ECM in the same manner as cell extracts, HK media PlEl anti-epilrqrin monoclonal antibody (ilAb) and MAb GB3. Studies of other patients with the same clinical phenotype found that while seven have anti-EM (i.e., autoantibodies (IqG, N=S:IqA, N=x), these autoantibodies are directed exclusively against the epidermal side of present In low titer (i.e., < 1:12), 1 M NaCl split skin, nonreactive with HK ECM, and negative in IP studies of HK These findings indicate that CP media and cell extracts. is a disease phenotype in which patlent autoantibodies tarclet different BM constituents and that anti-epillgrin autoantibodies are a specific marker for a unique form of Cp that we propose to call anti-epiliqrin Cp.
M.
Welch.
CP)(N=10)