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1991 Fiscal Year Final Research Report Summary

Molecular biological studies on a hepatoma-associated target antigen for a human killer T-cell clone

Research Project

Project/Area Number 01480222
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Gastroenterology
Research InstitutionUniversity of Tokyo

Principal Investigator

IMAWARI Michio  University of Tokyo Faculty of Medicine Assistant professor, 医学部(病), 助教授 (70134228)

Co-Investigator(Kenkyū-buntansha) NAKAMURA Ikuo  University of Tokyo Faculty of Medicine Fellow, 医学部(病), 医員
GUNJI Toshiaki  University of Tokyo Faculty of Medicine Fellow, 医学部(病), 医員
NAKAGAMA Hitoshi  University of Tokyo Faculty of Medicine Associate, 医学部(病), 助手
Project Period (FY) 1989 – 1991
Keywordshepatoma / killer T-cell / tumor-associated antigen
Research Abstract

We previously identified a tumor-associated target antigen, ATM-1, for a human killer T-cell clone 5B5, and found the existence of ATM-1 in sera of patients with hepatocellular carcinoma. To clone the gene that coded ATM-1, we screened the CDNA library obtained from a human hepatocellular carcinoma cell line, SK-HEP-1, immunologically using monoclonal antibodies raised against partially purified ATM-1. However we could not succeed in the cloning of ATM-1 gene possibly due to the low specificity and low sensitivity of the monocional antibodies used. Thus we changed the strategy and tried to purify the ATM-1 and determine the N-terminal amino acid sequence of the ATM-1. We purified ATM-1 from sera of patients with hepatocellular carcinoma by gel filtration on Sepharose CL-6B followed by affinity chromatography on anti-ATM-1 -bound Sepharose beads. As a control, serum from a healthy person was processed in the same way. Purified samples were subjected to SDS-polyacrylamide gel electrophoresis, and the separated proteins were transferred to a nitrocellulose membrane electrophoretically and stained. The sample from sera of patients with hepatocellular carcinoma gave a protein band with a molecular wright of approximately 55, 000 that was not observed in the sample from serum of a healthy person. On N-terminal amino acid sequence analysis, the protein band was found to contain two proteins. On homology search, one protein was found to have a sequence of immunoglobulin heavy chain hypervariable region III, and the other protein did not have any known amino acid sequence. We thought that the immunoglobulin was purified as an immune complex with ATM-1. We will raise antibodies against the synthetic peptide for the N-terminal amino acid sequence of the unknown protein, and also synthesize oligomers for the amino acid sequence. We will clone the ATM-1 gene using these antibodies and oligomers. Then we will study the biological and clinical significance of ATM-1.

  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] Kaieda T.: "Identification of a tumor-associated target antigen,ATM-1,for a human T-cell clone with sctivated killer activity and its existence in sera of cancer patients" Cancer Res. 48. 4848-4854 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 中釡 斉: "ヒトキラ-T細胞が認識する新しい肝癌マ-カ-ATM1ー17の物性解析とその肝癌における発現の検討." 第9回腫瘍マ-カ-研究会記録. 14-16 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kaieda T, et al.: "Identification of a tumor-associated target antigen, ATM-1, for a human T-cell clone with activated killer activity and its existence in sera of cancer patients." Cancer Res. 48. 4848-4854 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hitoshi Nakagama, et al.: "Physico-chemical analyses of a new tumor marker, ATM1-17, for hepatocellular carcinoma recognized by a human killer T-cell clone, and studies on the expression of the antigen in hepatoma tissues." Proc of the 9th Meeting on Research of Tumor Markers. 14-16 (1989)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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