Establishment of domain specific Enzyme-Linked ImmunoSorbent Assay for pemphigus using recombinant proteins expressed by mammalian cells.

• Project/Area Number: 25860973
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Dermatology
• Research Institution: Kurume University
• Principal Investigator: OHYAMA Bungo 久留米大学, 医学部, 講師 (90461441)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 自己免疫性水疱症 / エピトープ解析 / ドメイン特異的ELISA法 / 哺乳類細胞発現系組み換え蛋白質 / 昆虫細胞発現系組み換え蛋白質 / 免疫沈降法 / 免疫ブロット法 / ELISA法 / バキュロウイルス発現系 / 哺乳細胞発現系

Outline of Final Research Achievements

Pemphigus is autoimmune blistering disease caused by autoantibodies. Autoantibodies react with extracellular domains of desmoglein (Dsg) 1, Dsg3, desmocollin 1, and Dsc3 as antigens, and then cause blisters by abolished the adhesion between Dsg and Dsc proteins.
We attempted to establish domain specific Enzyme-Linked ImmunoSorbent Assay for pemphigus using recombinant proteins expressed by mammalian cells, because recombinant proteins of mammalian expression system keeps higher-order structures of protein than baculovirus insect expression system. We released some interesting data about epitopes which autoantibodies reacted against Dsg EC domains in three reports, and have established of IgA ELISAs of mammalian recombinant proteins of human Dsc 1-3.

Research Products

• [Journal Article] Establishment of IgA ELISAs of mammalian recombinant proteins of human desmocollins 1-3. (2016)
  • Author(s): eye K, Numata S, Ohzono A, Ohyama B, Tsuchisaka A, Koga H, Hachiya T, Tsuruta D, Ishii N, Hashimoto T
  • Journal Title: J Dermatol Sci Volume: 印刷中 Issue: 1 Pages: 75-77
  • DOI: 10.1016/j.jdermsci.2016.04.001
  • Peer Reviewed
• [Journal Article] Clinical and immunological profiles in 17 Japanese patients with drug-induced pemphigus studied at Kurume University. (2014)
  • Author(s): Yoshimura K, Ishii N, Hamada T, Abe T, Ono F, Hashikawa K, Fukuda S, Ohyama B, Koga H, Sogame R, Teye K, Ochiai T, Nakajima H, Nakajima K, Iijima S, Kanzaki M, Kojima K, Nagatani T, Fujimoto W, Karashima T, Nakama T, Ohata C, Furumura M, Tsuruta D, Hashimoto T
  • Journal Title: Br J Dermatol Volume: 171 Issue: 3 Pages: 544-553
  • DOI: 10.1111/bjd.12925
  • Peer Reviewed
• [Journal Article] Detection of Autoantibodies to Precursor Proteins of Desmogleins in Sera of a Patient with Bowen Carcinoma (2014)
  • Author(s): Demitsu T, Yamada T, Nakamura S, Kakurai M, Dohmoto T, Kamiya K, Aoyama Y, Iwatsuki K, Yamagami J, Ohyama B, Ohata C, Koga H, Hashimoto T
  • Journal Title: Acta Derm Venereol Volume: 94 Issue: 5 Pages: 601-603
  • DOI: 10.2340/00015555-1776
  • Peer Reviewed
• [Journal Article] Antibodies to the amino-terminal domain of desmoglein 1 are retained during transition from pemphigus vulgaris to pemphigus foliaceus. (2014)
  • Author(s): Espana A, Koga H, Suarez-Fernandez R, Ohata C, Ishii N, Irarrazaval I, Teye K, Ohyama B, Hashimoto T
  • Journal Title: Eur J Dermatol Volume: in press
  • Peer Reviewed