Preliminary study of HCV mouse model induced by infectious HCV 2a plasmid
-
摘要:
目的制备表达丙型肝炎病毒(HCV)2a型RNA的小鼠模型,为研究HCV高亲和力中和单克隆抗体在体内的病毒清除能力提供可行的动物模型。方法采用引进的HCV 2a型的质粒Jc1,体外表达HCV RNA后转染Huh7细胞,建立可持续感染的Huh7-Jc1丙肝细胞模型。然后以该细胞腹腔注射裸鼠,或以该细胞尾静脉注射裸鼠,并通过小鼠血液中HCV 2a的RNA水平检测,观察小鼠体内的HCV存留情况,以移植未转HCV的Huh7细胞和培养上清为阴性对照。结果建立了可表达HCV RNA和核心蛋白的Huh7-Jc1细胞,且该细胞模型具有一定感染能力,以其培养上清孵育Huh7细胞,可使后者也表达HCV RNA和核心蛋白。BALB/c-nu裸鼠腹腔或尾静脉移植Huh7-Jc1细胞后,裸鼠血清中可检测到HCV RNA的表达。结论显示利用Huh7-Jc1细胞可初步制备表达HCV的丙型肝炎小鼠模型,但高效、长久表达HCV的小鼠模型还有待进一步完善。
Abstract:Objective To establish a mouse model which can produce hepatitis C virus(HCV) RNA for examination of the efficiency of virus clearance of high-affinity HCV monoclonal antibody in vivo.Methods Huh7-Jc1 cell model was established by transfecting a genotype 2a HCV plasmid Jc1 into Huh7 cell line which can produce HCV RNA in vitro.The Huh7-Jc1 cell was injected into the nude mouse abdominally or intravenously and/,and the remaining HCV RNA level was detected by optical in vivo imaging and virus RNA in peripheral blood was also detected.Negative control was established by injecting non-HCV Huh7 cell.Results A HCV cell model was estabished which could express HCV RNA and core protein.and the supernatant of the HCV cell model also had the ability to infect the Huh7 cells.And the HCV RNA could be detected in the serum of the BALB/c-nu nude mouse which were implanted with Huh7-Jc1 cell(intraperitoneal or tail intravenous).Conclusion Our data indicates that implanting Huh7-Jc1 cell is a feasible method for building a HCV expressing mouse model,however,a higher efficiency and long-term virus producing mouse model still needed moreresearch.
-
Key words:
- hepacivirus /
- disease models
-
[1]Sarbah SA,Younossi ZM.Hepatitis C:an update on the silent epidemic[J].J Clin Gastroenterol,2000,30(2):125-143. [2]Hu YW,Rocheleau L,Larke B,et al.Immunoglobulin mimicry by Hepatitis C Virus envelope protein E2[J].Virology,2005,332(2):538-549. [3]Tarr AW,Owsianka AM,Timms JM,et al.Characterization of the hepatitis C virus E2 epitope defined by the broadly neutralizing monoclonal antibody AP33[J].Hepatology,2006,43(3):592-601. [4]Tokushige K,Wakita T,Pachuk C,et a1.Expression and lmmume Response to Hepatitis C Virus Core DNA—Base Vaccine Constructs[J].Hepatology,1996,24(1):l4-20. [5]Guévin C,Lamarre A,LabontéP.Novel HCV replication mouse model using human hepatocellular carcinoma xenografts[J].Antiviral Res,2009,84(1):14-22. [6]Lindenbach BD,Evan MJ,Syder AJ,et al.Complete replication of hepatitis C virus in cell culture[J].Science,2005,309(5734):623-626. [7]Zhu N,Ware CF,Lai MM.Hepatitis C virus core protein enhances FADD-mediated apoptosis and suppresses TRADD signaling of tumor necrosis factor receptor[J].Virology,2001,283(2):178-187. [8]Chou AH,Tsai HF,Wu YY,et al.Hepatitis C virus core protein modulates TRAIL-mediated apoptosis by enhancing Bid cleavage and activation of mitochondria apoptosis signaling pathway[J].J Immunol,2005,174:2160-2166.
计量
- 文章访问数: 2925
- HTML全文浏览量: 13
- PDF下载量: 1017
- 被引次数: 0