2000 Fiscal Year Annual Research Report

BP230とBP180のリコンビナント蛋白を用いた自己免疫水疱症の解析

Research Project

Project/Area Number	11670857
Research Institution	Kurume University School of Medicine
Principal Investigator	駒井礼子久留米大学, 医学部, 助手 (70281532)
Co-Investigator(Kenkyū-buntansha)	橋本隆久留米大学, 医学部, 教授 (20129597)
Keywords	自己免疫性水疱症 / 水疱性類天疱瘡 / ヘミデスモソーム / リコンビナント蛋白 / 免疫ブロット法 / BP180 / BP230 / ELISA
Research Abstract	昨年度はBP180cDNAの4つの部分(細胞外部分全長をカバーする3部位およびNC16a部位)のリコンビナント蛋白を作製し、水疱性類天疱瘡と瘢痕性類天疱瘡の反応を検討した。本年度は、さらに、C末端から15番目のコラーゲン部位(C15)をコードするcDNAを、特異的プライマーと正常ケラチノサイトcDNAライブラリーを用いたPCR法により得た。そのcDNAを大腸菌発現ベクターpGEXに組込み、大腸菌にトランスフェクトし、リコンビナントのGST融合蛋白を誘導した。その蛋白を用いた免疫ブロット法で、線状IgA水疱性皮膚症の血清がこのリコンビナント蛋白に特異的に反応することを見いだした。この事実は、線状IgA水疱性皮膚症が、BP180分子上の特異的なエピトープに反応することを示唆した。同様に、特異的プライマーと正常ケラチノサイトcDNAライブラリーを用いたPCR法により、全長のBP230分子cDNAをカバーするほぼ同じ大きさ(約3kb)の異なった3つのcDNAを得た。そのcDNAを大腸菌発現ベクターpGEXに組込み、大腸菌にトランスフェクトし、GST蛋白とのリコンビナント融合蛋白を得た。その蛋白を用いた免疫ブロット法で、多くの水疱性類天疱瘡が各部位のリコンビナント蛋白、とくC末端部位に特異的にまた強く反応することが判明した。古典的天疱瘡や腫瘍随伴性天疱瘡患者血清の一部もこれらのBP230の部位特異的リコンビナント蛋白にわずかに反応したが、その反応は水疱性類天疱瘡に比べて非常に弱いものであった。この事実はBP230が水疱性類天疱瘡特異的抗原であることを示唆した。さらに、以前作製したBP180 NC16a部位のリコンビナント蛋白を用いたELISAを作製し、現在、水疱性類天疱瘡を含めた各種患者血清の反応を検討中である。今後従来の蛍光抗体法に代わる検査法になる可能性がある。

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] Hashimoto T et al: "Continuing Medical Education : The new pemphigus variants"J Am Acad Dermatol. 40. 649-671 (1999)
[Publications] Hashimoto T et al: "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)
[Publications] Komai A,Hashimoto T et al: "Desmoglein 1 and desmoglein 3 are the target autoantigens in herpetiform pemphigus"Arch Dermatol. 135. 943-947 (1999)
[Publications] Hashimoto T et al: "IgA antibodies of cicatricial pemphigoid sera specifically react with C-terminus of BP180"J Invest Dermatol. 112. 254-255 (1999)
[Publications] Hashimoto T et al: "Paraneoplastic pemphigus with cutaneous and serological features of pemphigus foliaceus"Br J Dermatol. 141. 351-359 (1999)
[Publications] Komai A,Hashimoto T et al: "Sublamina densa-type linear IgA bullous dermatosis successively treated with oral tetracyclin and niacinamide"Br J Dermatol. 141. 608-609 (1999)
[Publications] Hashimoto T et al: "Unique immunobullous disease in a child with a predominantly IgA response to three desmosomal proteins"Br J Dermatol. 141. 882-886 (1999)
[Publications] Komai A,T.Hashimoto et al: "Pemphigoid nodularis with IgA autoantibodies against the intracellular domain of desmoglein 1"Br J Dermatol. 142. 143-147 (2000)
[Publications] Hashimoto T,Komai A et al: "Bullous pemphigoid associated with silicosis"Dermatology. 201. 265-267 (2000)
[Publications] Hashimoto T et al: "Rapid response of IgA pemphigus of subcorneal pustular dermatosis type to treatment with isotretinoin."J Am Acad Dermatol. 43. 923-926 (2000)
[Publications] Hashimoto T et al: "Autoantibodies in anti-p200 pemphigoid stain skin lacking laminin 5 and type VII collagen"Br J Dermatol. 143. 1043-1049 (2000)
[Publications] Hashimoto T et al: "IgA antibodies of linear IgA bullous dermatosis recognize the 15th collagenous domain of BP180"J Invest Dermatol. 115. 1164-1166 (2000)
[Publications] Komai A,Hashimoto T et al: "Anti-desmoglein autoantibodies in silicosis patients with no bullous diseases"Dermatology. (in press).
[Publications] Hashimoto T et al: "Clinical significance of enzyme-linked immunosorbent assay for the detection of circulating anti-BP180 autoantibodies in patients with bullous pemphigoid"J Dermatol Sci. (in press).
[Publications] Hashimoto T,Komai A et al: "Immunopathological diagnosis of cicatricial pemphigoid with desquamative gigivitis-A case report"J Periodontology. (in press).
[Publications] Komai A,Hashimoto T et al: "The clinical transition between pemphigus foliaceus and pemphigus vulgaris correlates well with the changes in autoantibody profile assessed by an enzyme-linked immunosorbent assay"Br J Dermatol. (in press).
[Publications] Hashimoto T,Komai A et al: "Desmogleins are targeted by IgA autoantibodies of a few IgA pemphigus patients"Arch Dermatol. (in press).
[Publications] Hashimoto T et al: "Bullous pemphigoid sera react specifically with various domains of BP230, most frequently with C-terminal domain, by immunoblot analysis using bacterial recombinant proteins covering the entire molecule"Exp Dermatol. (in press).

2000 Fiscal Year Annual Research Report

BP230とBP180のリコンビナント蛋白を用いた自己免疫水疱症の解析

Principal Investigator

駒井 礼子 久留米大学, 医学部, 助手 (70281532)

Research Products

[Publications] Hashimoto T et al: "Continuing Medical Education : The new pemphigus variants"J Am Acad Dermatol. 40. 649-671 (1999)

[Publications] Hashimoto T et al: "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)

[Publications] Komai A,Hashimoto T et al: "Desmoglein 1 and desmoglein 3 are the target autoantigens in herpetiform pemphigus"Arch Dermatol. 135. 943-947 (1999)

[Publications] Hashimoto T et al: "IgA antibodies of cicatricial pemphigoid sera specifically react with C-terminus of BP180"J Invest Dermatol. 112. 254-255 (1999)

[Publications] Hashimoto T et al: "Paraneoplastic pemphigus with cutaneous and serological features of pemphigus foliaceus"Br J Dermatol. 141. 351-359 (1999)

[Publications] Komai A,Hashimoto T et al: "Sublamina densa-type linear IgA bullous dermatosis successively treated with oral tetracyclin and niacinamide"Br J Dermatol. 141. 608-609 (1999)

[Publications] Hashimoto T et al: "Unique immunobullous disease in a child with a predominantly IgA response to three desmosomal proteins"Br J Dermatol. 141. 882-886 (1999)

[Publications] Komai A,T.Hashimoto et al: "Pemphigoid nodularis with IgA autoantibodies against the intracellular domain of desmoglein 1"Br J Dermatol. 142. 143-147 (2000)

[Publications] Hashimoto T,Komai A et al: "Bullous pemphigoid associated with silicosis"Dermatology. 201. 265-267 (2000)

[Publications] Hashimoto T et al: "Rapid response of IgA pemphigus of subcorneal pustular dermatosis type to treatment with isotretinoin."J Am Acad Dermatol. 43. 923-926 (2000)

[Publications] Hashimoto T et al: "Autoantibodies in anti-p200 pemphigoid stain skin lacking laminin 5 and type VII collagen"Br J Dermatol. 143. 1043-1049 (2000)

[Publications] Hashimoto T et al: "IgA antibodies of linear IgA bullous dermatosis recognize the 15th collagenous domain of BP180"J Invest Dermatol. 115. 1164-1166 (2000)

[Publications] Komai A,Hashimoto T et al: "Anti-desmoglein autoantibodies in silicosis patients with no bullous diseases"Dermatology. (in press).

[Publications] Hashimoto T et al: "Clinical significance of enzyme-linked immunosorbent assay for the detection of circulating anti-BP180 autoantibodies in patients with bullous pemphigoid"J Dermatol Sci. (in press).

[Publications] Hashimoto T,Komai A et al: "Immunopathological diagnosis of cicatricial pemphigoid with desquamative gigivitis-A case report"J Periodontology. (in press).

[Publications] Komai A,Hashimoto T et al: "The clinical transition between pemphigus foliaceus and pemphigus vulgaris correlates well with the changes in autoantibody profile assessed by an enzyme-linked immunosorbent assay"Br J Dermatol. (in press).

[Publications] Hashimoto T,Komai A et al: "Desmogleins are targeted by IgA autoantibodies of a few IgA pemphigus patients"Arch Dermatol. (in press).

[Publications] Hashimoto T et al: "Bullous pemphigoid sera react specifically with various domains of BP230, most frequently with C-terminal domain, by immunoblot analysis using bacterial recombinant proteins covering the entire molecule"Exp Dermatol. (in press).

駒井礼子久留米大学, 医学部, 助手 (70281532)