| Project/Area Number |
62480199
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Keio University |
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Principal Investigator |
MORIZANE Toshio School of Med. Keio Univ. Internal Med Lecturer, 医学部・内科学, 専任講師 (20129703)
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| Co-Investigator(Kenkyū-buntansha) |
IWABUCHI Naoto Scool of Med. Keio Univ. Internal Med., Assistant, 医学部・内科学, 助手 (90168592)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1989: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Hepatocellular carcinoma / Monoclonal antibody / Tumor-specific antigen / Tumor-specific antibody / 1-butanol extract / HCCM / 1ーbutanol extract / tumor-sepecific antigen / モノクローナル抗体 / 癌化 / 血清診断 / 肝特異抗原 / 肝細胞がん / 細胞膜抗原 / 肝細胞 |
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| Research Abstract |
We have attempted to detect antibodies against tumor-specific antigens (TSA) of hepatocellular carcinoma (HCC) in sera from patients with HCC using a human hepatoma cell line, HCC-M, which we had established in our laboratory. we have also attempted to produce reurine monoclonal antibodies reactive with this cell line. Cell surface antigen molecules of HCC-M cells were extracted with 1-butanol extraction method from the suspension of living cells. An enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibodies to this extract. Antibody binding was detectable in sera from patients with liver cirrhosis (LC) as well as HCC. There was no statistically significant difference between these two diseases. However, after absorbing sera with a normal liver antigen preparation, it was possible to demonstrate HCC-specific antibody binding to the HCC-M butanol extract. Therefore, we thought that this antigen preparation contained TSA which was common in HCC.
we developed reurine monoclonal antibodies against the HCC-M butanol extract and obtained 1H-9 monoclonal antibody. It was demonstrated by immunoperoxidase technique and ELISA that 1H-9 monoclonal antibody was reactive with HCC-M cells, HCC tissues but not with cancer cells of other types. The reactivity of 1H-9 with normal liver cells was almost negligible. We developed an ELISA for determining the serum level of the corresponding antigen. It was higher in 19 out of 26 cases of HCC than normal range. This result suggests that the determination of the serum level of the antigen recognized by 1H-9 monoclonal antibody serves ad a diagnostic tool for HCC. The 1-butanol extract of HCC-M cells was divided into 17 fractions by a high pressure liquid chromatography and immune reactivities evaluated by leukocyte adherence inhibition assay and antibody binding assay were demonstrable in a small molecule fraction. These results also suggest that the 1-butanol extract of HCC-M contains TSA of HCC.
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