| Project/Area Number |
13670879
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | Nagoya University |
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Principal Investigator |
MURO Yoshinao Nagoya University, Graduate School of Medicine Associate Professor, 大学院・医学系研究科, 助教授 (80270990)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Atopic Dermatitis / Epitope / Anti-DFS70 Antibodies / Alopecia / Recombinant protein / ELISA / 坑DFS70抗体 |
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| Research Abstract |
1. I established ELISA for quantitative analysis of anti-DFS70 antibodies using highly purified recombinant protein expressed in E. coli.
2. According to results of immunoblotting with bacterial recombinant protein and immunoprecipitation with in vitro translation and transcription product, it was suggested that some anti-DFS70 antibodies could recognize the comformational structure of the protein.
3. Epitope mapping studies with various lengthes of recombinant protein showed that the antibodies reacted to a single portion of the protein with little heterogeneity and that several dozens of amino acid residues were necessary to form the epitope. The negative data on epitope mapping studies with synthetic peptides derived from 12 amino acid sequences also suggested that the autoepitope of DFS70 is protein conformation dependent.
4. Anti-DFS70 antibodies were also found in some patients with alopecia and healthy individuals.Titers of the antibodies were higher in patients with atopic dermatitis and/or alopecia than in healthy individuals.
5. Polyclonal rabbit antibodies against DFSF70 were successful to be produced with synthetic peptides derived from the C-terminal sequence of the protein, which were applicable to immunostaining of cultured cells.
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