| Project/Area Number |
11670536
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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 |
Gastroenterology
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| Research Institution | Jikei University School of Medicine |
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Principal Investigator |
ZENIYA Mikio JIKEI UNIVERSITY SCHOOL OF MEDICINE, DIVIGION OF GASTROENTEROLOGY AND HEPATOLOGY, DEPARTMENT OF INTERNAL MEDICINE, ASSISTANT PROFESSOR, 医学部, 助教授 (70138767)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Hiroki JTKEI UNIVERSITY SCHOOL OF MEDICINE, DIVIGION OF GASTROENTEROLOGY AND HEPATOLOGY, DEPARTMENT OF INTERNAL MEDICINE, ASSISTANT, 医学部, 助手 (80256403)
渡辺 文時 東京慈恵会医科大学, 医学部, 助手 (90231711)
AIZAWA Yoshio JIKEI UNIVERSITY SCHOOL OF MEDICINE, DIVIGION OF GASTROENTEROLOGY AND HEPATOLOGY, DEPARTMENT OF INTERNAL MEDICINE, ASSITANT PROFESSOR, 医学部, 講師 (90147273)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | AUTOIMMUNE HEPATITIS / ANIMAL MODEL / DENDRITIC CELL / 肝内T細胞浸潤 / HLA-DR4 / CD25 / AIH / DR4 / 樹状細胞 / 肝癌細胞 / 融合細胞 / 実験的自己免疫性肝炎 / 細胞障害性T細胞 |
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| Research Abstract |
Backaround: We tried to establish a newly animal autoimmune hepatitis (AIH) model by using the fusion cell of hepatocyte and dendritic cell (DC) to investigate the immimopathogenesis of AIH.
Methoth: DC were generated by 5-days culture of bone marrow cells of C57BL/6 female mice with IL-4 and GM-CSF and fusion with well differentiated hepatoma cell Hepal-6 (H-2Db) were carried out in the presence of 50% PEG. Fusion cells were injected to C57BL/6 mice twice a month subcutaneously followed by peritoneal injection of IL-12 (500ng/body) three times a week. Splenocytes were extracted after final IL-12 injection and cultured with IL-2 (50 μg/ml) for 4 days. cytotoxicity of splenocytes against Hepal-6 was analyzed using 4hr 51Cr release assay. As a control, splenocytes obtamed from non treated, IL-12 treated, fusion cells treated mice were studied.
Results: Splenocytes extracted from fusion cell treated mice showed Hepa 1-6 specific cytotoxicity (51% cytolysis, E/T ratio80: 1), but not against
… More
NK cell sensitive Yac-1 cell. T cell population, which were separated using mierobeades and MACS magnetic separation system, of splenocytes obtained from fusion cell treated mice showed Hepa 1-6 specific cytotoxicity (41% cytolysis, EIT ratio=80: 1). But non T cell population did not show cytotoxicity Interestingly, splenocytes extracted from fusion cell and IL-12 treated mice did not show Hepa 1-6 specific cytotoxicity. Splenocytes extracted from non-treated mice nor IL- 12 treated mice didn t show cytotoxicity either. Immuno-histochemical analysis revealed that intrahepatic T cell infiltration was observed in fusion cell and IL-12 treated mice, but not in fusion cell treated mice.
Conclusion: These findings indicate that immunization with fusion cells of dendritic cell and Hepa 1-6 may evoke cytotoxic T cell which recoginize the epitope that is shared between Hepa 1-6 and hepatocyte in vivo, but it is not enough to induce hepatocyte injury. IL-12 may participate in the mechanisms of intrahepatic CTL infiltration and hepatocyte injury in this model. Less
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