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1989 Fiscal Year Final Research Report Summary

Evaluation of High-molecular Weight Epidermal Growth Factor for Its Distribution in Human and Animal Tissues

Research Project

Project/Area Number 63480204
Research Category

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

Allocation TypeSingle-year Grants
Research Field Gastroenterology
Research InstitutionNagoya City University Medical School

Principal Investigator

ITOH Makoto  Nagoya City Univ. Medical School Associate Professor, 医学部, 助教授 (00080119)

Co-Investigator(Kenkyū-buntansha) YASUE Naoji  Nagoya City Univ. Medical School Associate Professor, 医学部(平成元年度研究分担者), 助手 (80210243)
MIYAMOTO Tadahisa  Nagoya City Univ. Medical School Associate Professor, 医学部(昭和63年度研究分担者), 助手 (90182047)
YOKOYAMA Yoshifumi  Nagoya City Univ. Medical School Associate Professor, 医学部, 助手 (00145723)
Project Period (FY) 1988 – 1989
KeywordsHigh Molecular Weight Epidermal Growth Factor / EGF / Enzyme Immunoassay / Immunohistochemistry / EGF Precursor / EGF前駆体
Research Abstract

To assess in detail the physiological distribution of epidermal growth factor, attempts were made to measure the contents of high molecular weight human EGF (HMW-hEGF) in a variety of tissues in man and animals by establishing an enzyme immunoassay (EIA) system, and to immunohistochemically detect its localization in the tissues.
(1) Preparation of EIA system for HMW-hEGF: In the amino acid sequence of HMW-hEGF lying at 828-1023 on the NH2-terminal side of the molecular structure of HEGF precursor, two segments of polypeptide (amino acids 858-873 and 952-969) were synthesized and coupled to bovine serum albumin or hemocyanin.
These four substances were emulsified with Freund's adjuvant and injected sc into separate groups of the New Zealand albino rabbits. Eighteen weeks later, two anti-HMW-hEGF antisera to the polypeptides were obtained and their r-globulin fractions prepared. Using these antibodies and anti-hEGV- antibody which we had already possessed, sandwich enzyme immunoassay syst … More ems comprising the antibody (Fab')2- immobilized solid-phase and the antibody Fab' fractions labeled with 3-D-galactosidase were prepared.
Standard HMW-hEGF was provided by Wakunaga Pharmaceuticals Co., in Japan. Contrary to expectation, the maximal intensity of fluorescence was limited to 3 times the control. For the time, further studies are planned to establish an EIA system using synthesized polypeptides composing of higher molecules than those used in the present study.
(2) Purification of HMW-hEGF: We purified about 300 ng of HMW-hEGF from 290 g of HMW-hEGF-containing urine material eluted and concentrated from 330 1 of human urine (provided by Hitachi Chemical Co.).
However, HMW-hEGF over 10-20 times as much amount as obtained in the present study is required for the preparation of an EIA system. Since the collection of urine needed for the purification of such a lot of HMW-hEGF is difficult, the attempt to purify HMW-hEGF was judged to be impossible.
(3) Histochemistry using HMW-hEGF Antibodies: Using the 858-872 and 952-969 antibodies, HMW-HEGF was assessed for its distribution in human and animal tissues by employing the ABC method. Both the 858-872 and 952-969 antibodies showed negative staining in all the rat tissues tested: the submandibular gland, pancreas, liver, kidney, stomach, duodenum and the colon. However, the 952-969, but not the 858-872, antibody resulted in positive staining in interstitial and intimal cells and uriniferous tubule cells of the kidney and in ductal and interstitial cells of the submandibular gland in man. A study for identifying the substances stained by the antibody with HMW-hEGF is in progress. Less

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Makoto IToh,Takashi Toh,Shinpei Imai,Tadahisa Miyamoto.Ken Matsusaka et al.: "Experimenthland chinical studiee on epidermalgrowthfactor for gastric mawsal protecticn and healing of gassiriculars." J.Clin Gastroenterol.10 suppl.1. S7-S12 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 宮本忠寿,伊藤誠,横山善文,安江直二,今井新平ほか: "家兎胃粘膜臓器管培養法によるペプシトゲン分泌の検討-Epidermal Gropwth Factor(EGF)の抑制効果について-" 日本消化器病学会雑誌. 85. 1077-1081 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤誠: "EGFの胃粘膜防御,修復作用と抗潰瘍剤としての展望" 医薬ジャ-ナル. 24. 2667-2672 (1988)

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      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤誠,城卓志,今井新平,宮本忠寿,安江直二,武井俊彦: "胃粘膜防御機構としてのEpidermal Gropwth Factor" クリニカ. 15. 348-354 (1988)

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      「研究成果報告書概要(和文)」より
  • [Publications] Takashi Joh,Makoto Itoh,Naojiyaseu,Tadahisa Miyamoto,Akira Iwai,Ken Matsusaka et al.: "a sensitive enjyme immunoassay system of rat epidermalgrowth fador in biologicul fluids and tissue extracts." Acta Endoctinol. 120. 616-623 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤誠: "Epidermal growth Factor" 消化性潰瘍、基礎臨床. 8. 178-189 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤誠(分担執筆): "胃粘膜防御の仕組-あゆみと展望-、EGF" 竹本忠良ほか編集、医歯薬出版社, 9 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 伊藤誠,安江直二,武井俊彦(分担執筆): "胃潰瘍の治療と再発防止-臨床の最前線- 胃潰瘍とEGF、ロイコトリエン、SOD" 岡部治弥ほか編集、医薬ジャ-ナル社, 9 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Makto Itoh, Takashi Joh, Shinpei Imai, Tadahisa Miyamoto, Kei Matsusako, Akira Iwai, Kohei Katsumi, Kazuo Endoh, Kazuo Goto and Toshihiko Takeuchi.: "Experimental and clinical studies on epidermal growth factor for gastric mucosal protection and healing of gastric ulcers." J. Clin. Gastroenterol 10 (Suppl. 1):S7-S12, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tadahisa Miyamoto, Makoto Itoh, Yoshifumi Yokoyama, Naoji Yasue, Shinpei Imai, Takashi Joh, Kazuo Ikeda, Akira Iwai, Kei Matsusako, Toshihiko Takeuchi, Yoshiki Noguchi, Kohei Katsumi and Kiyoshi Yokochi.: "Inhibitory effect of epidermal growth factor on pepsinogen secretion in rabbit gastric mucosa." Gastroenterol. Jpn. 85:1077-1081, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Itoh.: "Effect of EGF on gastric mucosal protection and healing of gastric ulcers - An outlook of EGF for an anti-ulcer drug -." Iyaku Journal, 24:2667-2672, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Itoh, Takashi Joh, Shinpei Imai, Tadahisa Miyamoto, Naoji Yasue and Toshihiko Takeuchi.: "Role of epidermal growth factor in the mechanism of gastric mucosal protection." Clinica 15:348-354, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takashi Joh, Makoto Itoh, Naoji Yasue, Tadahisa Miyamoto, Akira Iwai, Kei Matsusako, Toshihiko Takeuchi, Akihiko Moriyama, Taiji Kato and Ryo Tanaka.: "A sensitive enzyme immunoassay system of rat epidermal growth factor in biological fluids and tissue extracts." Acta Endocrinol. 120:616-623, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Itoh.: "Prospect of EGF as a New Anti-ulcer Drug." Clinica 16:222-228, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Itoh.: "Epidermal Growth Factor" Peptic Ulcer: Kiso to Rinsyo 8:178-189, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kei Matsusako, Makoto Itoh, Takashi Joh, Tadahisa Miyamoto, kiyoshi Yokochi, Toshihiko Tadeuchi, Akihiko Moriyama, Taiji Kato and Ryo Tanaka.: Clin. Chimi. Acta 181:19-26, 1989.

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Published: 1993-03-26  

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