| Project/Area Number |
11671553
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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 |
Urology
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| Research Institution | Kobe University Graduate School of Medicine |
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Principal Investigator |
HARA Isao Kobe University Graduate School of Medicine Lecturer, 大学院・医学系研究科, 講師 (10263378)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAI Hiroshi Kobe University Graduate School of Medicine, Assistant, 大学院・医学系研究科, 助手 (80335447)
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| Project Period (FY) |
1999 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | IL-18 / Immune gene therapy / renal cell carcinoma |
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| Research Abstract |
Interleukin 18 (IL-18) is a novel cytokine identified as a strong interferon g inducer. IL-18 has similar bioactivities as interleukin-12 (IL-12) and demonstrates anti-tumor effects. Since IL-18 does not have signal sequence, we constructed the gene, consisting of the signal sequences of interferon b and mature IL 18 cDNA. The modified gene was subsequently transduced into mouse renal cell carcinoma cell line to induce IL-18 secretion from the tumor cells to establish whether this IL-18 secreting tumor cell line can induce systemic tumor immunity. Modified IL-18 cDNA consisting of the signal sequences of interferon b and mature type of IL-18 cDNA was constructed by the overlap extension method. The modified IL-18 cDNA and original IL-18 cDNA were transduced into mouse renal cell carcinoma cell line (RenCa). Expression of IL-18mRNA and concentration of IL-18 in the culture supernatant were determined. Direct anti-tumor effects and tumor vaccine effects were investigated in syngeneic Balb/c mice. To determine the mechanism of the anti-tumor effect, immunodeficient mice were challenged with IL-18 secreting RenCa cells. Although the modified IL-18 and original IL-18 gene transduced RenCa showed almost the same level of IL-18 mRNA expression, only RenCa cells transduced with modified IL-18 gene secreted IL-18 into the culture supernatant and were completely rejected when transplanted into syngeneic mice. T lymphocytes were involved in this anti-tumor effect. Moreover, the modified IL-18 transduced RenCa induced tumor vaccine effect against parental RenCa cells injected at a distant site. In conclusion, Immune gene therapy using modified IL-18 cDNA appears to be effective, including that IL-18 secreting cancer cell may be a candidate for tumor vaccine therapy.
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