Hepatocarcinogenesis in transgenic mice carrying hepatitis C virus cDNA

携带丙型肝炎病毒 cDNA 的转基因小鼠的肝癌发生

• 批准号: 11470061
• 负责人: ESUMI Mariko
• 金额: $ 8.58万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470061/
• 关键词: Hepatitis C virus; Hepatocellular carcinoma; Transgenic mouse; Albumin promoter; SRα promoter; Srαプロモーター

1 Expression analysis of transgenic miceWe analyzed 5 lines of transgenic mice carrying the whole genome of hepatitis C virus (HCV); A44, A39 and A48 were controlled under the mouse albumin enhancer/ promoter; S2 and S5 were under the SR_α promoter. The HCV mRNA was expressed in the liver of one of 17 A44 at the age of 12 months and 12 of 17 A44 at the age of 21 months. All mice of A39 and A48 expressed the HCV transgene in the liver, but both S2 and S5 did not. The mRNA was 9.4 kb in length, and the real-time PCR revealed that 50 to 4000 copies of HCV RNA were expressed in 1 μg of total RNA. The expression of HCV mRNA seemed to be localized in the liver tissue. Less than the detection limit of HCV core protein (5 pg/mg protein) was detected in the liver. These transgenic mice A39, A44 and A48 showed the similar level of HCV expression to that of patients with chronic hepatitis C, that is different from other transgenic mice with the high level of HCV expression.2 Histological examinat … More ion of transgenic mouse liverInflammatory cell infiltration and spotty necrosis were observed in transgenic mouse liver of A39, A44 and A48. The number of spotty necrosis foci was significantly higher in A39 and A44 than non-transgenic mice at the age of 21 months. The fatty change of liver was not significant between transgenics and non-transgenics. Liver fibrosis and tumorous lesion were not observed until 21 months old. HCV antibodies were not detected in serum of the mice.3 Molecular pathogenesis of transgenic mouse liverReal-time RT-PCR of mRNA revealed that mRNAs such as interferon-inducible genes and cell cycle-related genes were up-regulated in transgenic mouse liver, that is observed in patients with HCV infection.Thus, these transgenic mice were similar to patients with weak hepatitis. Anti-viral immune response of the mice, however, was so much less than that of patients, suggesting that the introduction of anti-viral immune system to these HCV transgenic mice should be required for establishing a model of hepatocarcinogenesis. Less

1转基因小鼠的表达分析分析了携带丙型肝炎病毒（HCV）整个基因组的5行转基因小鼠。 A44，A39和A48在鼠标专辑增强器/启动子下受到控制； S2和S5在SR_α启动子下。 HCV mRNA在12个月的17个A44之一的肝脏中表达，在21个月大时，在17个A44中的12个A44中表达。 A39和A48的所有小鼠均表示肝脏中的HCV转基因，但S2和S5都没有。 mRNA的长度为9.4 kb，实时PCR显示50至4000份HCV RNA拷贝在1μg的总RNA中表达。 HCV mRNA的表达似乎位于肝组织中。在肝脏中检测到HCV核蛋白（5 pg/mg蛋白）的检测极限。这些转基因小鼠A39，A44和A48表现出与慢性丙型肝炎患者的HCV表达水平相似的水平，这与其他HCV表达高水平的转基因小鼠不同。2组织学检查…更多的转基因小鼠脂肪蛋白炎性细胞浸润和斑点Necrisos的离子在转生小鼠中均在转基因小鼠中观察到A49的44.39。在A39和A44中，斑点坏死灶的数量明显高于21个月时的非转基因小鼠。肝脏的脂肪变化在转化体和非转基因学之间并不显着。直到21个月大，才观察到肝纤维化和肿瘤病变。 3在小鼠的血清中未检测到HCV抗体。3mRNA的转基因小鼠LiveReal-lime rt-PCR的分子发病机制表明，在TrygenIS中，在Tregnic semice中观察到了Trymatic ipnic MICIA，在转染中观察到了与转基因小鼠肝脏中的MRNA，例如干扰素诱导基因和细胞周期相关的基因。然而，小鼠的抗病毒免疫响应比患者少得多，这表明应该需要将抗病毒免疫系统引入这些HCV转基因小鼠，以建立肝癌发生模型。较少的

项目成果

Iwasaki Y, Esumi M., Hosokawa N., Yanai M. and Kawano K.: "Occasional infection of hepatitis C virus occurring in haemodialysis units identified by serial monitoring of the virus infection"J. Hosp. Infect.. 45(1). 54-61 (2000)

Iwasaki Y、Esumi M.、Hosokawa N.、Yanai M. 和 Kawano K.：“通过对病毒感染的系列监测发现血液透析单位偶尔发生丙型肝炎病毒感染”J.

Zhou Y, et al.(2): "Multiple sequence-reactive antibodies induced by a single peptide immunization with hypervariable region 1 of hepatitis C virus"Virology. 262. 360-370 (1999)

Zhou Y等人(2)：“丙型肝炎病毒高变区1单肽免疫诱导的多重序列反应性抗体”病毒学。

Esumi M, et al.(4): "In vivo and in vitro evidence that cross-reactive antibodies to C-terminus of hypervariable region 1 do not neutralize heterologous hepatitis C virus"Vaccine. 20. 3095-3103 (2002)

Esumi M 等人 (4)：“体内和体外证据表明，高变区 1 C 末端的交叉反应抗体不会中和异源丙型肝炎病毒”疫苗。

Zhou Y, Moriyama M. and Esumi M: "Multiple sequence-reactive antibodies induced by a single peptide immunization with hypervariable region 1 of hepatitis C virus"Virology. 256(2). 360-370 (1999)

Zhou Y、Moriyama M.和Esumi M：“丙型肝炎病毒高变区1单肽免疫诱导的多种序列反应性抗体”病毒学。

Esumi M, et al.(6): "Experimental vaccine activities of recombinant E1 and E2 glycoproteins and hypervariable region 1 peptides of hepatitis C virus in chimpanzees"Archives of Virology. 144. 973-980 (1999)

Esumi M等人(6)：“丙型肝炎病毒重组E1和E2糖蛋白和高变区1肽在黑猩猩中的实验疫苗活性”病毒学档案。