1999 Fiscal Year Final Research Report Summary
Identification of antigen for cause of ITP by the new molecular biological method (SEREX)
| Project/Area Number |
10670943
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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 |
Hematology
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| Research Institution | Mie University |
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Principal Investigator |
WADA Hideo Mie University, Faculty of Medicine, assistant, 医学部, 助手 (40158704)
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| Project Period (FY) |
1998 – 1999
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| Keywords | ITP / SEREX / cDNA / specific antibody / sequence / UT-7 / Molecular biology / recombinant protein |
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| Research Abstract |
To identify the cause of specific antigen in idiopathic thrombocytopenic purpura (ITP), we directly analyzed the gene of specific antigen in ITP by high sensitivity of genetic cloning method ; serological identification of antigens by recombinant expression cloning (SEREX). We extracted the RNA from UT-7/thrombopoietin and made the cDNA. After expression of protein from the cDNA in E-Coli, 38 clones which were reacted to the serum of patients with ITP were obtained. Out of them, 9 clones were unknown and 19 were known by the analysis of sequence of gene. We focused human hepatitis delta antigen interacting protein A (dip A), because it strongly reacted to the ITP serum. By RT-PCR method, we detected the expression of dip A in all cell line. We examined the anti-dip A antibody in patients with ITP by western blot method. Sixty percent of ITP patients were positive for dip A antibody, but only 20% of healthy volunteers were positive. The possibility of point mutation in dip-A and role of anti-dip-A antibody in the onset of ITP should be studied furthermore.
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