2003 Fiscal Year Final Research Report Summary
The establishment of diagnostic system for all the autoimmune bullous disease using various biochemical techniques
| Project/Area Number |
13557072
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Dermatology
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| Research Institution | KURUME UNIVERSITY |
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Principal Investigator |
HASHIMOTO Takashi Kurume University, Dermotol, Professor, 医学部, 教授 (20129597)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMA Takekuni Kurume University, Dermotol, As Prof, 講師 (50221453)
YASUMOTO Shinichiro Kurume University, Dermotol, As Prof, 助教授 (10220162)
MORI Osamu Kurume University, Dermotol, As Prof, 助教授 (10175630)
INOUE Mitsuse Kurume University, Dermotol, As Prof, 講師 (10232563)
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| Project Period (FY) |
2001 – 2003
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| Keywords | Autoimmune disease / Pemphigus / Bullous pemphigoid / Recombinant protein / Immunoblotting / ELISA / Desmosome / Hemidesmosome |
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| Research Abstract |
We isolate cDNA of desmocollin (Dsc) 1-3, and produced baculoproteins of Dsc1-3 using vaculovirus expression system. Using these recombinant protein, we established ELISA of Dsc. We produced bacterial recombiant proteins of the 3 different domains of envoplakin and periplakin using pGEX expression system, and detected autoantibodies in paraneoplastic pemphigus sera by immunoblotting using the recombinant proteins. We also produced recombinant proteins of 3 different domains of BP230, and detected autoantibodies in bullous pemphigoid by immunoblotting using the proteins. We produced recombinant proteins of various domains of BP18O, and examined the reactivity of cicatricial pemphigoid by immunoblotting using the recombinant proteins. We established immunoblotting using laminin 5 purified from the keratinocyte cultures and detected autoantibodies in the sera of anti-laminin 5 cicatricial pemphigoid. We established immunoblotting using concentrated culture supernatant of keratinocytes, and detected antibodies against LAD1 in LAD sera. We produced pGEX recombinant proteins of NC1 and NC2 domains of type VII collagen, and found that most EBA sera reacted with NC1 domain, and a few EBA sera reacted with NC2 domain.
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