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Immunohistochemical and biochemical analysis of antigenetic determinants on neuroendocrine tumors using monoclonal antibody HISL-19

Research Project

Project/Area Number 07671342
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General surgery
Research InstitutionNippon Medical School

Principal Investigator

SHIMIZU Kazuo  Nippon Medical School, Department of Surgery II,Associate Professor, 医学部, 助教授 (20133449)

Co-Investigator(Kenkyū-buntansha) TANAKA Shigeo  Nippon Medical School, Department of Surgery II,Professor, 医学部, 教授 (70089720)
WATANABE Hidehiro  Nippon Medical School, Department of Surgery II,Clinical instructor, 医学部, 助手 (40191788)
IWAMURA Taro  Nippon Medical School, Department of Surgery II,Clinical instructor, 医学部, 助手
NAGAHAMA Mitsuji  Nippon Medical School, Department of Surgery II,Clinical instructor, 医学部, 助手 (30241102)
名取 穣二  日本医科大学, 医学部, 助手
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsmonoclonal antibody / endocrine tumor / HISL-19 / medullary carcinoma of the thyrod / pheochromocytoma / pancreatic islet cell tumor / tumor marker / immunohistochemistry / モノクローナル抗体HISL-19 / 神経内分泌腫瘍 / 内分泌細胞癌 / カルチノイド / 免疫組織化学 / Western Blotting法
Research Abstract

[Introduction]
We studied immunohistochemical and biochemical analysis on neuroendocrine and non-neuroendocrine tumors using various kinds of monoclonal antibodies such as A2B5,4F2, PKK-1 and HISL-19. Among these antibodies, A2B5 and HISL-19 showed similar reactivity to only neuroendocrine cells and their tumors in immunohistochemical investigation. HISL-19, especially, obtained numerously remarkable outcomes because of the advantages that it can vbe applied for formalin-fixed, paraffin-embedded permanent specimens. We, herein, report our results of 2 years study with the HISL-19 as central figure.
[Materials and Methods]
1) Immunohistochemical study : Materials were mainly used formalin-fixed, paraffin embedded surgical specimens of neuroendocrine, and non-neuroendocrine tumors. Indirect immunofluorescence andor SAB procedures were applied for ataining.
2) Biochemical study : Cellular protein from 1 g of cryopreserved surgical specimen of medullary carcinoma of the thyroid (MCT) obtained … More at the time of surgery were extracted. Western blotting procedure was applied to determine the molecular weight of HISL-19 antigenic determinant.
3) Immunoelectron microscopic study : Conventional paraffin specimens for light microscope were first immunostained with HISL-19. Positive area was selected and applied for electron microscopic study. Electron microscopic specimens were processed to pre-embedding procedure and viewed with electron microscope with lead staining or no staining.
[Results]
1) HISL-19 reacted only with MCT in the thyroid neoplasms. In adrenal tumors, pheochromocytoma was positive while adrenocortical tumors were negative to it. In Pancreas, islet cell tumors were positive while pancreatic acinar cells were negative. In gut organ, carcinoid, gastric cell carcinoma were negative, but other esophageal, gastric, rectal, colonic carcinoma were negative. Interestingly, pathologically diagnosed endocrine cell carcinoma from gall bladder were positive to HISL-19. Malignant pheochromocytoma and MCT revealed strongly positive conpared with benign pheochromocytoma in virtue of SAB procedure.
2) Biochemical analysis using Western blotting procedure revealed new antigen HISL-19 band around 42 kDa in benighn pheochromocytoma and MCT which is immunohistochemically positive to HISL-19. Furtheremore, other two bands with mulecular weitght of 55 and 65 kDa were recognized in MCT which showed similar reactivity to malignant pheochromocytoma and did not show in benighn pheochromocytoma.
3) Immunoelectron microscopic procedure demonstrated HISL-19-positive substances which numerously exist in the secretory granules in cytoplasm of the MCT.These positive granules were densely aggregated closed to nucleus and showed a tendency of getting scattered toward apical surface of the tumor cells.
[Summary]
Monoclonal antibody HISL-19 is an clinically useful tool as a apecific marker for neuroendocrine cell tumors described above. Furthermore, malignant endocrine cell tumors appeared as newly synthesized protein. These were demonstrated as two different molecule bands such as 55 and 60 kDa which were not recognized in benign pheochromocytoma (HISL-19-positive : molecular weight of 42 kDa) and other HISL-19-negative tumors. These HISL-19 positive-proteins were located in nuerous secretory granules suggesting unfeasible cell regulation producing endless cell proliveration. Clinically, preoperative immunohistochemical diagnosis using HISL-19 proved to be claryfy the grade of malignancy, estimate the prognosis, make a decision of operative prcedure including extent of resection and lymphnode dissection. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report

Research Products

(14 results)

All Other

All Publications (14 results)

  • [Publications] Kazuo Shimizu,et al: "Immunohistochemical,biochemical and immunoelectron microscopic analysis on neuro-endocrine tumors using HISL-19" Fifth International Congress on Hormone and Cancer at Quebec City (Proceedings). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kazuo Shimizu,et al: "Monoclonal antibody HISL-19 : Its immunohistochemical biochemical and immunoelectron microscopic charaderistics for N-E cells and tumors" 9th annual meecting of the Asia-Pacific Endocrine Conference (Proceedings). 40-40 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Shimizu,et al: "Clinicopathological Study of Clear-cell Tumors of the Thyroid : An Evaluation of 22 Cases." Surg. Today. 25. 1015-1022 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Shimizu,et al: "Study on the Improvement of Thyroid Function after Autotransplantation of Cryopreserved Thyroid Tussue in Rets" Thyroidology,Clin. Exp. 8. 55-62 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, K., et al.: "Immunohistochemical, biochemical and immunoelectron microscopic analysis on neuro-endocrine tumors using monoclonal antibody HISL-19" Fifth International Congress on Hormone and Cancer at Quebec City. (Proceedings). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, K., et al.: "Monoclonal antibody HISL-19 : Its immunohistochemical, biochemical and immunohistochemical characteristics for neuro-endocrine cells and tumors" 9^<th> annual meeting of the Asia-Pacific Endocrine Conference. (Proceedings). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, K., et al.: "Clinicopathological Study of Clear-Cell Tumors of the Thyroid : An Evaluation of 22 Cases" Surg Today. 25. 1015-1022 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shimizu, K., et al.: "Study on the Improvement of Thyroid Function after Autotransplantation of Cryopreserved Thyroid Tissue in Rats" Thyroidology, Clin Exp. 8. 55-62 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kazuo Shimizu,et al.: "Immunohistochemical,biochemical and immunoelection microscopic analysis on neuro-endocrene tumors using HISL-19" Fifth International Congress on Hormone and Cancer at Quebec City (Proceedings). (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kazuo Shimizu,et al.: "Monoclonal antibody HISL-19 : Its immunohistochemical,biochemical and immunoelection microscopic characteristics for N-E cells and For N-E Cells tumors" 9th Annual Meeting of the ASIA-PACIFIC ENDOCRINE CONFERENCE (Proceedings). 40-40 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Shimizu,et al.: "Clinicopathological Study of Clear-Cell Tumors of the Thyroid : An Evaluation of 22 Cases" Surg.Today. 25. 1015-1022 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Shimizu,et al.: "Study on the Improvement of Thyroid Function after Autotransplantation of Ouyopreseeved Thyroid tissue in Rat-subtitle" THYROIDOLOGY,Clin Exp. 8. 55-62 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kazuo Shimizu et al: "monoclonal anttbody HISL-19:Its immunohistochemical,bicchemical and unmuno-electron microscopic characteristica for neuro-endocrine cells and their tumors." 9th Annual meeting of the ASIA-PACIFIC ENDOCRINE CONFERENCE (Proceedings). 40-40 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kazuo Shimizu et al: "lmmunohistochemical,biochemical and immunoelectron micro-scopic analysis on neuro-endocrine tumors using HISL-19" Fifth International Congress on Hormone and Cancer at Quebec city (Proceedings). (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-03-31   Modified: 2016-04-21  

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