| Project/Area Number |
03671156
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
NAGAFUCHI Seiho Kyushu Univ.Fac.Med.Assistant Prof., 歯学部, 助手 (00150441)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Minoru Kyushu Univ.Fac.Med.Assist.Prof., 歯学部, 助手 (40217906)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | diabetes / NOD mouse / insulitis / B lymphocytes / Glutamic acid Decarboxylase / anti-Phospholipid Ab / immunoglobulin subclass / autoimmunity / Glutamic Acid docarboxylase / GAD / インスリン |
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| Research Abstract |
We studied the infiltrating lymphocytes to islets and circulating autoantibodies in NOD mice. Anti-glutamic acid decarboxylase (GAD) antibody was produced in 46/99(46.5%) mice. Mice produced the anti-GAD antibody 39.5% at the age of 5-15 weeks and 96.9% at the age of 26 to 35 weeks, thus indicating production of the autoantibody was elevated along with the aging and diabetes occurred.The specificity of anti-GAD antibody was determined by the inhibition assay,inhibited by GAD and not inhibited by insulin. The immunoglobulin subclass of anti-GAD antibody was IgG2a. Anti-phospholipid antibody was produced at the rate of 37.4%. Mice produced anti-phospholipid antibody 6.9% at the age of 5 to 15 weeks,31.6% at the age of 16 to 25 weeks,and 71.9% at the age of 26 to 35 weeks.The antibody level was elevated according to the aging. Moreover,mice with diabetes possessed anti-phospholipid antibody at higher rate(84.6%) and mice without diabetes had the autoantibody at lower rate(63.2%).The specificity of the autoantibody was ensured by the specific inhibition by cardiolipin but not by insulin nor ssDNA. The immuoglobulin subclass of anti-phospholipid antibody was also IgG2a. We also studied the infiltrating lymphocytes by immunohistochemical study. At the early stage,we observed CD5 and CD4 positive T lymphocytes with scattering CD8 positive cells but no NK cells nor B lymphocytes were infiltrated.Along with the progression of insulitis,B lymphocytes were accumulated around CD4 positive T cells. Those B cells expressed IgM and IgG immunoglobulins on their surfaces but IgA positive cells were scarcely detected.The surface immunoglobulin subclass of infiltrated B lymphocytes was initially IgG2a and then IgG3 positive cells increased predominantly. We are now analysing in vitro of infiltrated B cells, whetter they produce autoantibodies as in vivo.The genetic analysis of repertoire of immnoglobulin V regions of infiltrated B lymphocyte are currently underway.
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