2002 Fiscal Year Final Research Report Summary
ANALISES OF PROPAGATION PROCESS OF HEPATITIS C VIRUS IN CULTURED LIVER CELL AND APPLICATIONS FOR ANTI-VIRUS DRUGS.
| Project/Area Number |
12670477
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Mie University |
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Principal Investigator |
KAITO Masahiko MIE UNIVERSITY, FACULTY OF MEDICINE, ASSISTANT PROFESSOR, 医学部, 講師 (70214244)
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| 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)
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| Project Period (FY) |
2000 – 2002
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| Keywords | HEPATITIS C VIRUS / CELL CULTURE / ELECTRON MICROSCOPE / IMMUNOELECTRON MICROSCOPY / IMMUNOHISTOCHEMISTRY / VIRUS VECTOR / LACTOFERRIN / IRON RESTRICTION |
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| 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.
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Research Products
(17 results)
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[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
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[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
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