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ANALISES OF PROPAGATION PROCESS OF HEPATITIS C VIRUS IN CULTURED LIVER CELL AND APPLICATIONS FOR ANTI-VIRUS DRUGS.

Research Project

Project/Area Number 12670477
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Gastroenterology
Research InstitutionMie University

Principal Investigator

KAITO Masahiko  MIE UNIVERSITY, FACULTY OF MEDICINE, ASSISTANT PROFESSOR, 医学部, 講師 (70214244)

Co-Investigator(Kenkyū-buntansha) KOHARA Michinori  TOKYO METROPOLITAN INSTITUTE OF MEDICAL SCIENCE, HEAD OF DEPARTMENT OF MICROBIOLOGY AND CELL BIOLOGY, 感染生体防御研究部門, 室長 (10250218)
WATANABE Shozo  MIE UNIVERSITY, STUDENTS HEALTH CENTER, PROFFESSOR, 保健管理センター, 教授 (20134934)
IKOMA Jiro  MIE UNIVERSITY, HOSPITAL, RESEARCH ASSOCIATE, 医学部附属病院, 助手 (80283521)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsHEPATITIS C VIRUS / CELL CULTURE / ELECTRON MICROSCOPE / IMMUNOELECTRON MICROSCOPY / IMMUNOHISTOCHEMISTRY / VIRUS VECTOR / LACTOFERRIN / IRON RESTRICTION / 鉄製限食
Research Abstract

A hybrid cell line (IMY) derived from human hepatocytes and HepG2 cells and that is highly sensitive to hepatitis C virus (HCV) infection was recently established. We also established a HCV replication system by using the entire HCV cDNA and T7 RNA polymerase recombinant adeno system. To detect HCV particles in the IMY cell culture supernatant, an immunoelectron microscopy of cell culture supernatant was carried out using monoclonal and polyclonal antibodies specific to the HCV structural proteins. The highest HCV RNA titer (3xl0^7 copies/ml) was found at the density between 1.12 and 1.15 g/ml of sucrose fractions. Morphological features of HCV particles in the culture supernatant are perfectly identical to those found in plasma samples from HCV-infected blood donors. HCV core particles are spherical particles of 33 to 40 nm in diameter and with an icosahedral symmetry. Separation and purification of HCV may enable the development of HCV vaccine and antiviral drugs. Moreover, the characteristic intracellular propagation of HCV has not been as yet evaluated in infected cells. To assess the intracellular propagation of HCV, we carried out immunoelectron microscopy in IMY cells. HCV core particles, of 30 to 35 nm in diameter, are localized in the cytoplasm. HCV particles, of 50 nm in diameter, were localized in cisternae of dilated endoplasmic reticulum and vacuoles. This study would allow us to clarify the mechanism of viral pathogenesis. We reported these research results in The American Association for the Study of Liver Diver Diseases (AASLD) 51^<ST> and 52^<ST> Annual Meeting.
We also reported in Hepato-Gastroenterology, 49(44) : 537-539, 2002 and American Journal of Gastroenterology. 97 : 3. 766-767, 2002 that lactoferrin therapy and iron restriction thepapy were effective for chronic hepatitis C patients.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (34 results)

All Other

All Publications (34 results)

  • [Publications] Motoh Iwasa, Masahiko Kaito, Jiro Ikoma, Shozo Watanabe et al.: "Dietary Iron Restriction Improves Aminotransferase Levels in Chronic Hepatitis C Patients"Hepato-Gastroenterology. 49(44). 537-839 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Jiro Ikoma, Masahiko Kaito, Shozo Watanabe, et al.: "Early Diagnosisi of Hepatocellular Carcinoma Using a Sensitive Assay for Serum Des-γ-Carboxy Prothrombin : a Prospective Study"Hepato-Gastroenterology. 49(43). 235-238 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Motoh Iwasa, Masahiko Kaito, Jiro Ikoma, Shozo Watanabe et al.: "Lactoferrin inhibits hepatitis C virus (HCV) viremia in chronic hepatitis C patients with a high viral load and HCV genotype 1b"American Journal of Gastroenterology. 97(3). 766-767 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Naoki Fujita, Masahiko Kaito, Jiro Ikoma, Shozo Watanabe, et al.: "Non-Immune Complexed HCV RNA Titer in Serum as a Predictor of Interferon Response in Patients with Chronic Hepatitis C. Gastroenterology"American Journal of Gastroenterology. 98(3)(In press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 垣内雅彦: "IMY細胞上清中のHCV粒子に関する免疫電子顕微鏡的研究"西部肝臓フォーラム 01'記録集編集 原田尚/谷川久一. 122-137 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 垣内雅彦, 岩佐元雄, 足立幸彦他: "難治性C型慢性肝炎に対するUDCA併用鉄制限食療法とラクトフェリン療法"消火器科 特集II「難治性C型慢性肝炎の治療法」. (In press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masahiko Kaito, Naoki Fujita, Jiro Ikoma, Shozo Watanabe, et al.: "Hepatitis C Virus Particles Detected by Immunogold Electron Microscopy"Science, Technology and Education of Microscopy, an Overview. A. Mendez Vilas編集(In press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Naoki Fujita, Masahiko Kaito, Satoshi Ishida, Naoki Nakagawa, Jiro Ikoma, Yukihiko Adachi, and Shozo Watanabe: "Paraformaldehyde Protects of Hepatitis C Virus Particles During Ultracenrifugation"Journal of Medical Virology. 63. 108-116 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Satoshi Ishida, Masahiko Kaito, Michinori Kohara, KyoKo Tsukiyama-Kohara, Naoki Fujita, Jiro Ikoma, Yukihiko Adachi, Shozo Watanabe: "Hepatitis C virus core particle detected by immunoelectron microscopy and optical rotation technique"Hepatology Research. 20. 335-347 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Motoh Iwasa, Masahiko Kaito, Jiro Ikoma, Yoshinao Kobayashi, Yuji Tanaka, Kunihiro Higuchi, Keisuke Takeuchi, Kazuko Iwata, Shozo Watanabe and Yukihiko Adachi: "Dietary Iron Restriction Improves Aminotransferase Levels in Chronic Hepatitis C Patients"Hepato-Gastroenterology. 49(44). 537-539 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Jiro Ikoma, Masahiko Kaito, Tomoaki Ishihara, Naoki Nakagawa, Akira Kamei, Naoki Fujita, Motoh Iwasa,Shigenori Tamaki, Shozo Watanabe, Yukihiko Adachi: "Early Diagnosis of Hepatocellular Carcinoma Using a Sensitive Assay for Serum Des-γ-Carboxy Prothrombin : a Prospective Study"Hepato-Gastroenterology. 49(43). 235-238 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Motoh Iwasa, Masahiko Kaito, Jiro Ikoma, Masaki Takeo, Ichiro Imoto, Yukihiko Adachi, Koji Yamagucji, Reiko Koixumi, Susumu Teraguchi: "Lactoferrin inhibits hepatitis C virus (HCV) viremia in chronic hepatitis C patients with a high viral load and HCV genotype 1b"American Journal of Gastroenterology. 97:3. 766-767 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masahiko Kaito, Noriko Yasui-Kawamura, Motoh Iwasa, Yoshinao Kobayashi, Naoki Nakagawa, Naoki Fujita, Jiro Ikoma, Esteban C.Gabazza, Shozo Watanabe, Yukihiko Adachi: "Twice-a-day versus one-a-day interferon-β therapy in chronic hepatitis C"Hepato-Gastroenterology. 50. 775-778 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Naoki Fujita, Masahiko Kaito, Masaki Takeo, Motoh Iwasa, Jiro Ikoma, Shozo Watanabe, and Yukihiko Adachi: "Non-Immune Complexed HCV RNA Titer in Serum as a Predictor of Interferon Response in Patients with Chronic Hepatitis C"The American Journal of Gastroenterology. 98(3)(in press). (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masahiko Kaito, Shozo Watanabe, Michinori Kohara, Kyoko Tsukiyama-Kohora, Satoshi Ishida, Masaki Takeo, Hideaki Tanaka, Shinichiro Horiike, Naoki Fujita, Motoh Iwasa, Esteban C Gabazza, Jiro Ikoma and Yukihiko Adachi: "Hepatitis C Virus Particles Detected by Immunogold Electron Microscopy"Science, Technology and Education of Microscopy : An Overview. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masahiko KAITO, Shozo WATANABE, Satoshi ISHIDA, Kyoko,TSUKIYAMA-KOHARA, Takayoshi ITO, Kotaro YASHUI, Naoki NAKAGAWA, Naoki FUJITA, Jiro IKOMA, Yukihiko ADACHI and. Michinori Kohara: "HEPATITIS C VIRUS PARTICLES DETECTED IN THE IMY CELL CULTURE SUPERNATANT BY IMMUNOELECTRON MICROSCOPY"Hepatology(AASLD ABSTRACTS). 32(4). 175-175 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masahiko KAITO, Shozo WATANABE, Kyoko TSUKIYAMA-KOHARA, Takayoshi ITO, Kotaro YASHUI, Masaki TAKEO, Naoki FUJITA, Jiro IKOMA, Yukihiko ADACHI and. Michinori Kohara: "PROPAGATION FEATURES OF HEPATITIS C VIRUS IN IMY CELL BY IMMUNOGOLD ELECTRON MICROSCOPY"Hepatology(AASLD ABSTRACTS). 34(4). 489-489 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Motoh lwasa, Masahiko Kaito, Jiro Ikoma, Shozo Watanabe et al.: "Dietary Iron Restriction Improves Aminotransferase Levels in Chronic Hepatitis C Patients"Hepato-Gastroenterology. 49(44). 537-839 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Jiro Ikoma, Masahiko Kaito, Shozo Watanabe, et al.: "Early Diagnosis of Hepatocellular Carcinoma Using a Sensitive Assay for Serum Des-γ-Carboxy Prothrombin : a Prospective Study"Hepato-Gastroenterology. 49(43). 235-238 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Motoh Iwasa, Masagiko Kaito, Jiro Ikoma, Shozo Watanabe et al.: "Lactoferrin inhibits hepatitis C virus (HCV) viremia in chronic hepatitis C patients with a high viral load and HCV genotype 1b"American Journal of Gastroenterology. 97(3). 766-767 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Naoki Fujita, Masahiko Kaito Jiro Ikoma, Shozo Watanabe, et al.: "Non-Immune Complexed HCV RNA Titer in Serum as a Predictor of Interferon Response in Patients with Chronic Hepatitis C. Gastroenterology"American Journal of Gastroenterology. 98(3)(In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 垣内雅彦: "IMY細胞上清中のHCV粒子に関する免疫電子顕微鏡的研究"西部肝臓フォーラム 01'記録集編集 原田 尚/谷川久一. 122-137 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 垣内雅彦, 岩佐元雄, 足立幸彦他: "難治性C型慢性肝炎に対するUDCA併用鉄制限食療法とラクトフェリン療法"消化器科 特集II 「難治性C型慢性肝炎の治療法」. (In press).

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiko Kaito, Naoki Fujita, Jiro Ikoma, Shozo Watanabe, et al.: "Hepatitis C Virus Particles Detected by Immunogold Electron Microscopy"Science, Technology and Education of Microscopy, an Overview. A. Mendez Vilas 特集(In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Satoshi Ishida, Masahiko Kaito, Michinori Kohara, Shozo Watanabe et al: "Hepatitis C virus core particle detected by immunoelectron microscopy and rotation technique"Hepatology Research. 20. 335-347 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Naoki Fujita, Masahiko Kaito, Jiro Ikoma, Shozo Watanabe et al.: "Paraformaldehyde protects of hepatitis C virus particles during ultacentrifugation"Journal of Medical Virology. 63・2. 108-116 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kaito Masahiko, Jiro Ikoma, Shozo Watanabe, Michinori Kohara et al: "Hepatitis C virus particles detected in the IMY cell culture supernatant by immunoelectron microscopy"Hepatology(AASLD ABSTRACTS). 32・4. 175-175 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Masaki Takeo, Masahiko Kaito, Motoh Iwasa, Naoki Fujita, et al.: "Lactoferrin inhibits hepatitis C virus(HCV)viremia in chronic hepatitis C patients with a high viral load and HCV genotype 1b"Hepatology(AASLD ABSTRACTS). 34・4. 331-331 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Masahiko Kaito, Shozo Watanabe, Kyoko Tsukiyama-Kohara, et al.: "Propagation feature of hepatitis C virus in IMY cell by immunogold electron microscopy"Hepatology(AASLD ABSTRACTS). 34・4. 489-489 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Motoh Iwasa, Masahiko Kaito, Yukihiko Adachi, et al.: "Lactoferrin inhibits hepatitis C virus(HCV)viremia in chronic hepatitis C patient with a high viral load and HCV genotype Ib"American Journal of Gastroenterology. 97・3. 766-767 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Satoshi Ishida,Masahiko Kaito,Michinori Kohara,Shozo Watanabe et al: "Hepatitis C virus core particle detected by immunoelectron microscopy and rotation technique."Hapatology Research. (In press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Naoki Fujita,Masahiko Kaito,Jiro Ikoma,Shozo Watanabe et al.: "Paraformaldehyde protects of hepatitis C virus particles during ultacentrifugation."Journal of Medical Virology. 63・2. 108-116 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kaito Masahiko,Jiro Ikoma,Shozo Watanabe,Michinori Kohara et al: "Hepatitis C virus particles detected in the IMY cell culture supernatant by immunoelectron microscopy."Hepatology (AASLDABSTRACTS). 32・4. 175-175 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Naoki Fujita,Masahiko Kaito,Jiro Ikoma,Shozo Watanabe et al.: "Clinical significance of serum levels of non-immune complexed hepatitis C virus in patients with chronic hepatitis C : Correlation with genotype and response to IFN therapy"Hepatology (AASLDABSTRACTS). 32・4. 288-288 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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