| Project/Area Number |
07670597
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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 | The Third Department of Internal Medicine, Ehime University School of Medicine |
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Principal Investigator |
ONJI Morikazu Ehime University School of Medicine Professor, 医学部, 教授 (10112260)
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| Co-Investigator(Kenkyū-buntansha) |
MASUMOTO Toshikazu Ehime University Hospital Assistant, 医学部・付属病院, 助手 (40243779)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | HCV / Lymphoid dendritic cells / Adenovirus vector / allogenic MLR / I-A / H-2K / C型肝炎 ウイルス / MHC class II / Lac Z遺伝子 |
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| Research Abstract |
Lymphoid dendritic cells (LDC) were transfected with HCV gene to clarify whether functional abnormality of murine LDC is related with persistent infection with hepatitis C virus (HCV) or not.
1.LDC (purity>80%) were isolated from C57BL/6 mice according to the method of Steinman-Inaba. These LDC were incubated with adenovirus vector transfected with LacZ gene (Adex1CALacZ) for 3 hours (moi>100) and the expression of LacZ were seen to be >80% on LDC by staining using X-gal.
2.LDC were transfected with adenovirus vector containing HCV gene, expanding between core and envelope region (Adex1CA327) and immunostaining using HCV core protein revealed that >80% LDC expressed HCV core protein.
3.LDC transfected with Adex1CA327 and control adenovirus vector (Adex1w1) were cultured with allogenic T lymphocytes in allogenic MLR and incorporation of [^3H] - Thymidine, expressing the level of DNA synthesis was compared. LDC,transfected with adenovirus vector containing HCV gene had significantly lower stimulatory capacity than the same transfected with control adenovirus vector (p<0.05). LDC transfected with HCV gene were also looked for the expressions of I-A,H-2K,B7-2, and ICAM-1, but no significant difference could be seen in the expressions of these molecules on LDC either being transfected with Adex1CA327 or Adex1w1.
From these observations, it was clarified that the stimulatory capacity of LDC was decreased due to transfection with HCV gene and the role of LDC in persistent HCV infection is postulated. The mechanism underlying this cellular dysfunction will be analyzed in future.
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