2002 Fiscal Year Final Research Report Summary
Establishment of tet-off HCV Transgenic mice and immunological response to DNA vaccination
| Project/Area Number |
12670520
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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 | Jikei University School of Medicene |
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Principal Investigator |
SATO Jujin Institute of Clinical Medicine and Research, Lecturer, 医学部, 講師 (30205893)
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| Co-Investigator(Kenkyū-buntansha) |
SANJO Akira Internal Medicine, Assistant, 医学部, 助手 (60301529)
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| Project Period (FY) |
2000 – 2002
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| Keywords | Hepatitis C virus / Transganic mouse / tet-off system / DNA vaccine |
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| Research Abstract |
As a prior step for establishing a small animal model to investigate HCV pathogenesis, we firstly established a binary transgenic model in which the conditional expression of lacZ as a reporter transgene can be tightly regulated in the fiver fey means of the tetracycline regulatory system. The mouse albumin promoter was used to achieve liver-specific expression of the teteaeycEne-responsive transcripa'onal activator (CTA) in one set of transgenic mice (Alfc-tTA). These mice were crossed with traasgenic mice carrying lacZ under the control of the tTA-regulated promoter (tetO-lac0 Analyses of mice transgenic for both tTA and lacZ (tTA/&zeZ> revealed thai the Ever-specific expression of the transgene could be suppressed to undetectable levels and regulated in a reversible fashion by tetracyeline administration and withdrawal.
Based on the successful establishment of the 0AJlacZ mice, tae binary transgenic mice carrying both tTA and full-length HCV CDNA (tTA/HCV) were generated by replacing
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lacZ with CDNA corresponding to the entire HCV open reading frame (strain Ib) in tetO-lacZ (tetO-HCV), followed by crossing tetQ-HCV wife Alb-tTA. TVo different hneages of the tTA/HCV have been obtained by screening integration of both HCV and tTA using Southern blot or PCR analysis. Furthermore, RT-PCR analysts demonstrated that HCV-specific RNA transeripts spanning the entire poiyprotem coding region, were induced exclusively in the livers in both of two lines by tefcacyeBae withdrawal. However, the levels of induced HCV core protein, measmed by higMy-sensttive EU」A. was definitely positive in one of two O1A/HCV fines.
These, indueible HCV transgenic mice can be used to observe not only the pathological consequences of HCV protein expression in the liver but the immunological responses against HCV gene products, especially whea the expressions are induced after birth. In addition,we are hopeful mat our model system wiU pave the way for the more precise study of any endogenous or exogenous genes and their interactions with the immune system in the context of viral immunopathology, autoimmunity, or other forms of liver disease. Less
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Research Products
(2 results)
