| Project/Area Number |
09670529
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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 | Shinshu University |
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Principal Investigator |
TANAKA Eiji Shinshu University School of Medicine, Lecturer, 医学部, 講師 (50163506)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | in-situ hybridization / liver tissue / hepatitis G virus / hepatitis C virus / chronic hepatitis / lymphocyte / 鎖特異的PCR / HCV / HGV / HGV-C / in-situ hybridization / 単核球 / HGV RNA / 一本鎖RNAプローブ / ジゴキシゲニン |
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| Research Abstract |
1. Condition of in-situ hybridization
Digoxigenine labeled RNA probe was prepared by in vitro transcription for in-situ hybridization. Nucleotide sequence of hybridization probe was selected from the NS5 region in both HCV and HGV. Hybridized probes were detected by DIG detection kit. Paraffin-embedded liver specimens were used in the present study.
2. Localization of HGV in liver tissue
Six patients with chronic HGV infection and 6 controls were analyzed. Positive signal of in-situ hybridization was observed in 2 of the 6 patients with HGV infection, while it was observed in none of the 6 controls. The 2 patients with the positive signal were under immuno-suppressive therapy after liver transplantation and had high level of HGV RNA in serum. Both positive and negative strands of HGV RNA were detected in mononuclear cells infiltrated in portal areas, but not in hepatocytes. Those results indicate that HGV replication is absent or limited in hepatocytes, and that HGV is not a hepatitis virus.
3. Localization of HCV in liver tissue Eight
patients with chronic hepatitis C and 8 controls were analyzed. Positive signal of in-situ hybridization was observed in all 6 patients with HCV infection, while it was observed in none of the 6 controls. The positive signal was observed mainly in hepatocytes (only in cytoplasm), and rarely in epitherium of bile ducts. Both positive and negative strands of HCV RNA were detected in hepatocytes. Hepatocytes with the positive signal tended to be more common in central part of lobule than in its peripheral part. Strength and localization of the positive signal did not differ between patients with low and high viral load. Clinical significance of HCV localization in liver tissue is now under investigation.
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