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Establishment of molecular biological tests for the diagnosis of various autoimmune bullous skin dermatosis with auto-basement manbrane zone autoantibodies

Research Project

Project/Area Number 09557065
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Dermatology
Research InstitutionKurume University

Principal Investigator

HASHIMOTO Takashi  Professor, Dermatology, Kurume University, 医学部, 教授 (20129597)

Co-Investigator(Kenkyū-buntansha) MIYASATO Minoru  Assistant Professor, Dermatology, Kurume University, 医学部, 講師 (50182001)
MOIRI Osamu  Associate Professor, Dermatology, Kurume University, 医学部, 助教授 (10175630)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1999: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥6,800,000 (Direct Cost: ¥6,800,000)
KeywordsAutoimmune / Keratinocyte / Basement membrane / Bullous pemphigoid / Hemidesmosome / Recombinant protein / PCR / Immunoblotting / 自己抗体 / 類天疱瘡 / 基底膜 / 分子生物学 / 免疫プロット法 / ELISA / cDNA
Research Abstract

We have obtained 3 cDNAs covering entire BP230 by PCR amplification using appropriate primers and cDNA library prepared from mRNA of cultured keratinocytes. We inserted these cDNAs into bacterial expression vector, pGEX, and produced recombinant fusion proteins to GST. Using immunoblotting of these recombinant proteins, we confirmed that bullous pemphigoid (BP) sera reacted with multiple epitopes on the BP230 molecule, and that this reactivity is specific to BP. By the similar method, we also obtained 3 recombinant GST fusion proteins, which covered the entire extracellular domain of BP180. Using immunoblotting of these recombinant proteins, we confirmed that BP sera specifically react with NC16a domain in the N-terminal region of BP180, and that cicatricial pemphigoid sera contain IgG and IgA reactive with C-terminal region of BP180. In addition, the sera of linear IgA bulloud dermatosis of lamina lucida type seem to reacti with epitope on the N-terminal region of BP180, which is a li … More ttle distal to C-terminal, if compared to the epitope for BP. Next, we obtained recombinant proteins of NC1 and NC2 domains of type VII collagen. By immunoblotting using these recombinant proteins, we found that both epidermolysis bullosa acquista and bullous SLE react preferentially with NC1 domain, and less frequently with NC2 domain of type Vli collagen. Linear IgA bullous dermatosis of sublamina densa type never reacted with either recombinant proteins. These immunoblot analyses using various different recombinant proteins of both BP180, BP230 and type VII collagen should be very important methods to obtain a definite diagnosis for various autoimmune bullous dermatosis with anti-basement membrane zone antibodies. We are now attempting to establish ELISA using these recombinant proteins, which will be much more convenient method for the diagnosis of these disease than the conventional immunofluorescence tests. Finally, we are now attempting to identify the 2000 kDa new deraml protein, which is reacted by some patients of psoriasis showing bullous lesions vesivular pemphigoid-like patients. The studies performed so far (cDNA cloning and various biological studies) revealed that this molecule may be related with type VII collagen or lamine gamma 1, or may be a new type of collagen. We will continue to work with this protein. Less

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (46 results)

All Other

All Publications (46 results)

  • [Publications] Murakami H et al.: "Analysis of antigens targeted by circulating IgG and IgA antibodies in 50 patients with cicatricial pemphigoid"J Dermatol Sci. 17. 39-44 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hirako Y et al.: "Cleavage of BP180 (Type XAII collagen) yields a 120 kDa collagenous extracellular peptide"J Biol Chem. 273(16). 9711-9717 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Chan LS et al.: "Epitope spreading : lessons from autoimmune skin diseases"J Invest Dermatol. 110. 103-109 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nousari HC et al.: "The mechanism of respiratory failure in paraneoplastic pemphigus"N Eng J Med. 340. 1406-1410 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nie Z, Hashimoto T: "Complementary peptide against the major epitope in NCa6A of BP180 show no spidicificity as vaccines ot bulous pemphigoid"J Dermatol Sci. 21. 157-164 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hashimoto T: "Skin diseases related to abnormality in desmosomes and hemidesmosomes"Dermatol Sci. 20. 81-84 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 橋本 隆: "KEY WORD皮膚疾患"先端医学. 210 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 橋本 隆: "皮膚科診療プラクティス"文光堂. 305 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Murakami H et al: "Analysis of antigents targeted by circulating IgG and IgA antibodies in 50 patients with cicatricial pemphigoid"J Dermatol Sci. 17. 39-44 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hirako Y et al: "Cleavage of BP180 (Type XVII collagen) yields a 120 kDa collagenous extracellular peptide"J Bio Chem. 273. 9711-9717 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Chan LS et al: "Epitope spreading : lessons form autoimmune skin diseases"J Invest Dermatol. 110. 103-109 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nousari HC et al: "The mechanism of respiratory failure in paraneoplastic pemphigus"N Eng J Med. 340. 1406-1410 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nie Z, Hashimoto T: "Complementary peptide against the major epitope in NCa6A of BP180 show no spidicificity as vaccines ot bulous pemphigoid"J Dermatol Sci. 21. 157-164 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hashimoto T: "Skin diseases related to abnormality in desmosomes and hemidesmosomes"Dermatol Sci. 20. 81-84 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Murakami H et al.: "Analysis of antigens targeted by circulating IgG and IgA antibodies in 50 patients with cicatricial pemphigoid"J Dermatol Sci. 17. 39-44 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hirako Y et al.: "Cleavage of BP180 (Type XVII collagen) vields a 120 kDa collagenous extracellular peptide"J Biol Chem. 273(16). 9711-9717 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] Chan LS et al.: "Epitope spreading: lessons from autoimmune skin diseases"J Invest Dermatol. 110. 103-109 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nousari HC et al.: "The mechanism of respiratory failure in paraneo;lastic pemphigus"N Eng J Med. 340. 1406-1410 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nie Z,Hashimoto T: "Complementary peptide against the major epitope in NCa6A of BP180 show no spidicificity as vaccines of bulous pemphigoid"J Dermatol Sci. 21. 157-164 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hashimoto T: "Skin diseases related to abnormality in desmosomes and hemidesmosomes"Dermatol Sci. 20. 81-84 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 橋本 隆: "KEY WORD皮膚疾患"先端医学. 210 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 橋本 隆: "皮膚科診療プラクティス"文光堂. 305 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kita Y: "T cell receptor clonotypes in skin lesions from patients with systemic lupus erythematosus" J Invest Dermatol. 110. 41-46 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Tanaka H: "A novel bullous dermatosis with autoantibodies against the collagenous domain of type VII collagen" Clin Invest. 77. 623-632 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hirako Y: "Cleavage of BP180(Type XVII collagen)yields a 120 kDa collagenous extracellular peptude" J Biol Chem. 273. 9711-9717 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Amagai M: "Antibodies against desmoglein 3(pemphigus vulgaris antigen)are present in sera from patients with paraneoplastic pemphigus and cause acantholysis in vivo in neonatal mice" J Clin Invest. 102. 775-782 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kiyokawa C: "Envoplakin and periplakin are components of the paraneoplastic pemphigus antigen complex" J Invest Deramtol. 111. 1236-1238 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nie Z: "IgA antibodies of cicatricial pemphigoid sera specifically react with C-terminus of BP180" J Invest Dermatol. 112. 254-255 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 橋本 隆: "今日の治療指針 1998年版" 医学書院, 1437 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 橋本 隆: "専門医のための皮膚科学レビュー" 総合医学社, 164 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ishikawa O: "The presence of anti-basement membrane zone antibodies in the sera of patients with non-bullous lupus" Br J Dematol. 136. 222-226 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kubo A: "Epidermolysis bullosa acquisita exacerbated by systemic estrogen and progesterone treatement and pregnancy" J Am Acad Dermatol. 36. 792-794 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hashimoto T: "Human desmocollin 1(Dsc1)is an autoantigen for subcorneal pustular dermatosis type of IgA pemphigus" J Invest Dermatol. 109. 127-131 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Okura M: "A case of vesicular pemphigoid with anti-desmoplakin autoantibodies" Br J Dermatol. 136. 794-795 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ishii K: "Charaterization of autoantibodies in pemphigus using antigen-specific enzyme-linked immunosorbent assays iwth baculovirus-expressed recombinant desmogleins" J Immunol. 159. 2010-2017 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hashimoto T: "Four cases of cicatricial pemphigoid showing IgG and IgA antibodies against the 180 kD bullous pemphigoid antigen" Br J Dermatol. 137. 305-306 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Endo Y: "Psoriasis vulgaris coexistent with epidermlysis bullosa acquisita" Br J Dermatol. 137. 783-786 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kubo A: "A case of herpetiform pemhigus showing reactivity with pemphigus vulgaris antigen(desmoglein 3)" Br J Dermatol. 137(1). 109-113 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kita Y: "T cell receptor clonotypes in skin lesious from patients with systemic lupus erythematosus" J Invest Dermatol. 110. 41-46 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hashimoto: "Novel non-radioisotope immunoprecipitation studies indicate involvement of pemphigus vulgaris antigen in paraneoplastic pemphigus" J Dermatol Sci. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Tanaka H: "A novel bullous dematosis with autoantibodies against the collagenous domain of type VII collagen" Clin Invest. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Hirako Y: "Cleavage of BP180(Type XVII collagen)yields a 120 kDa collagenous extracellular peptide" J Biol Chem. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] 橋本 隆: "皮膚科レジデント・戦略ガイド" 診断と治療社, 376 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 橋本 隆: "今日の治療指針 1998年版" 医学書院, 1437 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 橋本 隆: "皮膚診療クイックリファレンス" メジカルビュー社, 430 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 橋本 隆: "専門医のための皮膚科学レビュー" 総合医学社, 164 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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