| Project/Area Number |
18590994
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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 |
Metabolomics
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| Research Institution | Nagasaki University |
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Principal Investigator |
KAWASAKI Eiji Nagasaki University, Hospital of Medicine and Dentistry, Department of Metabolism/Diabetes and Clinical Nutrition, Associate Professor (70336171)
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| Co-Investigator(Kenkyū-buntansha) |
ABIRU Norio Nagasaki University, Graduate School of Biomedical Sciences, Department of Endocrinology and Metabolism, Assistant Professor (00380981)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,950,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2007: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Immunology / Type 1 diabetes / Autoantibody / Antigen recognition / Prediction / Protein / Genetics |
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| Research Abstract |
Japanese type 1 diabetes (T1D) is a heterogeneous disease characterized by the different clinical features in terms of its mode of onset or age at disease onset. It is estimated that the prevalence of adult-onset T1D is similar to that of childhood-onset T1D, and that around two-thirds of adult-onset T1D are categorized in "slow-onset T1D". Although anti-glutamic acid decarboxylase 65 (GAD65) antibodies are good predictive marker for future insulin deficiency in slow-onset T1D, positive predictive value at 5 years is about 65% and one-thirds of GAD65 antibody-positive patients do not need insulin therapy for long period. Therefore, in order to develop the highly predictive marker for future insulin deficiency in slow-onset T1D we performed a fine epitope mapping of anti-AD65 antibodies using random phage display technology.
1. Preparation of human islet GAD random phage display library
In the first year we prepared a human islet GAD random phage display library, and screened this library with pooled sera from patients with slow-onset T1D and healthy control subjects. After amplifying the individual clone that reacted with sera from patients or controls, the PCR-direct sequence analysis was performed to determine the specific DNA sequence for GAD65 molecule.
2. Development of anti-GAD epitope-specific antibody measurement
With the results obtained in the first year, we tried to identify the specific epitopes associated with the progression to insulin deficiency in slow-onset T1D patients by sandwich ELISA method. There were many difficulties to identify the diabetes-specific epitopes for GAD antibodies using random phage display library system with ELISA because of the high Noise/Signal ratio.
We are trying to do the fine epitope mapping using other detection systems.
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