期刊文献+
共找到38篇文章
< 1 2 >
每页显示 20 50 100
The woodchuck as an animal model for pathogenesis and therapy of chronic hepatitis B virus infection 被引量:23
1
作者 Stephan Menne Paul J Cote 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第1期104-124,共21页
This review describes the woodchuck and the woodchuck hepatitis virus (WHV) as an animal model for pathogenesis and therapy of chronic hepatitis B virus (HBV) infection and disease in humans. The establishment of wood... This review describes the woodchuck and the woodchuck hepatitis virus (WHV) as an animal model for pathogenesis and therapy of chronic hepatitis B virus (HBV) infection and disease in humans. The establishment of woodchuck breeding colonies, and use of laboratory-reared woodchucks infected with defined WHV inocula, have enhanced our understanding of the virology and immunology of HBV infection and disease pathogenesis, including major sequelae like chronic hepatitis and hepatocellular carcinoma. The role of persistent WHV infection and of viral load on the natural history of infection and disease progression has been firmly established along the way. More recently, the model has shed new light on the role of host immune responses in these natural processes, and on how the immune system of the chronic carrier can be manipulated therapeutically to reduce or delay serious disease sequelae through induction of the recovery phenotype. The woodchuck is an outbred species and is not well defined immunologically due to a limitation of available host markers. However, the recent development of several key host response assays for woodchucks provides experimental opportunities for further mechanistic studies of outcome predictors in neonatal- and adult-acquired infections. Understanding the virological and immunological mechanisms responsible for resolution of self-limited infection, andfor the onset and maintenance of chronic infection, will greatly facilitate the development of successful strategies for the therapeutic eradication of established chronic HBV infection. Likewise, the results of drug efficacy and toxicity studies in the chronic carrier woodchucks are predictive for responses of patients chronically infected with HBV. Therefore, chronic WHV carrier woodchucks provide a well-characterized mammalian model for preclinical evaluation of the safety and efficacy of drug candidates, experimental therapeutic vaccines, and immunomodulators for the treatment and prevention of HBV disease sequelae. 展开更多
关键词 woodchuck woodchuck hepatitis virus hepatitis B virus Neonatalacquired infection Adult-acquired infection Resolution CHRONICITY Humoral immune response Cellular immune response Antiviraltherapy Immunotherapy Combination therapy Hepatocellular carcinoma
下载PDF
Differentially expressed genes in hepatocellular carcinoma induced by woodchuck hepatitis B virus in mice 被引量:11
2
作者 Mark Feitelson 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第4期575-578,共4页
INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-... INTRODUCTIONHepatocellular carcinoma(HCC)is one of the major causes of death in the word.The mechanism of carcinogenesis is unknown,although it is widely accepted that HBV and HCV are clsely related to liver cancer[1-5[1-5].Previously,a variety of studies have described the differences in gene expression which distinguished tumor from nontumor[6-11].Cloning of the genes,especially the genes associated with HBV and HCV,is still very important to account for the development of liver cancer. 展开更多
关键词 Animals Carcinoma Hepatocellular Cloning Molecular DNA Complementary Databases Nucleic Acid Gene Expression Regulation Neoplastic Gene Expression Regulation Viral hepatitis B hepatitis B virus woodchuck Humans MICE Polymerase Chain Reaction Research Support Non-U.S. Gov't
下载PDF
Cross-species hybridization of woodchuck hepatitis virus-induced hepatocellular carcinoma using human oligonucleotide microarrays 被引量:1
3
作者 Paul W Anderson Bud C Tennant Zhenghong Lee 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第29期4646-4651,共6页
AIM: To demonstrate the feasibility of using woodchuck samples on human microarrays, to provide insight into pathways involving positron emission tomography (PET) imaging tracers and to identify genes that could be... AIM: To demonstrate the feasibility of using woodchuck samples on human microarrays, to provide insight into pathways involving positron emission tomography (PET) imaging tracers and to identify genes that could be potential molecular imaging targets for woodchuck hepatocellular carcinoma. METHODS: Labeled cRNA from woodchuck tissue samples were hybridized to Affymetrix U133 plus 2.0 GeneChips. Ten genes were selected for validation using quantitative RT-PCR and literature review was made. RESULTS: Testis enhanced gene transcript (BAX Inhibitor 1), alpha-fetoprotein, isocitrate dehydrogenase 3 (NAD+) beta, acetyI-CoA synthetase 2, carnitine palmitoyltransferase 2, and N-myc2 were up-regulated and spermidine/spermine N1-acetyltransferase was down-regulated in the woodchuck HCC. We also found previously published results supporting 8 of the 10 most up-regulated genes and all 10 of the 10 most downregulated genes. CONCLUSION: Many of our microarray results were validated using RT-PCR or literature search. Hence, we believe that woodchuck HCC and non-cancerous liver samples can be used on human microarrays to yield meaningful results. 展开更多
关键词 Cross-species hybridization Gene expression woodchuck hepatitis virus Hepatocellular carcinoma woodchuck Marmota monax
下载PDF
Application of the woodchuck animal model for the treatment of hepatitis B virus-induced liver cancer 被引量:1
4
作者 Manasa Suresh Stephan Menne 《World Journal of Gastrointestinal Oncology》 SCIE 2021年第6期509-535,共27页
This review describes woodchucks chronically infected with the woodchuck hepatitis virus(WHV)as an animal model for hepatocarcinogenesis and treatment of primary liver cancer or hepatocellular carcinoma(HCC)induced by... This review describes woodchucks chronically infected with the woodchuck hepatitis virus(WHV)as an animal model for hepatocarcinogenesis and treatment of primary liver cancer or hepatocellular carcinoma(HCC)induced by the hepatitis B virus(HBV).Since laboratory animal models susceptible to HBV infection are limited,woodchucks experimentally infected with WHV,a hepatitis virus closely related to HBV,are increasingly used to enhance our understanding of virus-host interactions,immune response,and liver disease progression.A correlation of severe liver pathogenesis with high-level viral replication and deficient antiviral immunity has been established,which are present during chronic infection after WHV inoculation of neonatal woodchucks for modeling vertical HBV transmission in humans.HCC in chronic carrier woodchucks develops 17 to 36 mo after neonatal WHV infection and involves liver tumors that are comparable in size,morphology,and molecular gene signature to those of HBV-infected patients.Accordingly,woodchucks with WHV-induced liver tumors have been used for the improvement of imaging and ablation techniques of human HCC.In addition,drug efficacy studies in woodchucks with chronic WHV infection have revealed that prolonged treatment with nucleos(t)ide analogs,alone or in combination with other compounds,minimizes the risk of liver disease progression to HCC.More recently,woodchucks have been utilized in the delineation of mechanisms involved in innate and adaptive immune responses against WHV during acute,self-limited and chronic infections.Therapeutic interventions based on modulating the deficient host antiviral immunity have been explored in woodchucks for inducing functional cure in HBV-infected patients and for reducing or even delaying associated liver disease sequelae,including the onset of HCC.Therefore,woodchucks with chronic WHV infection constitute a well-characterized,fully immunocompetent animal model for HBV-induced liver cancer and for preclinical evaluation of the safety and efficacy of new modalities,which are based on chemo,gene,and immune therapy,for the prevention and treatment of HCC in patients for which current treatment options are dismal. 展开更多
关键词 woodchuck hepatitis B virus Chronic infection Liver disease Hepatocellular carcinoma Cancer treatment
下载PDF
Animal models for the study of hepatitis B virus infection 被引量:16
5
作者 wei-na guo bin zhu +2 位作者 ling ai dong-liang yang bao-ju wang 《Zoological Research》 SCIE CAS CSCD 2018年第1期25-31,共7页
Even with an effective vaccine, an estimated 240 million people are chronically infected with hepatitis B virus (HBV) worldwide. Current antiviral therapies, including interferon and nucleot(s)ide analogues, rarel... Even with an effective vaccine, an estimated 240 million people are chronically infected with hepatitis B virus (HBV) worldwide. Current antiviral therapies, including interferon and nucleot(s)ide analogues, rarely cure chronic hepatitis B. Animal models are very crucial for understanding the pathogenesis of chronic hepatitis B and developing new therapeutic drugs or strategies. HBV can only infect humans and chimpanzees, with the use of chimpanzees in HBV research strongly restricted. Thus, most advances in HBV research have been gained using mouse models with HBV replication or infection or models with HBV-related hepadnaviral infection. This review summarizes the animal models currently available for the study of HBV infection. 展开更多
关键词 hepatitis B virus Animal model Duckhepatitis B virus woodchuck hepatitis virus
下载PDF
Potential mechanisms of hepatitis B virus induced liver injury 被引量:12
6
作者 Mohd Suhail Hany Abdel-Hafiz +5 位作者 Ashraf Ali Kaneez Fatima Ghazi A Damanhouri Esam Azhar Adeel GA Chaudhary Ishtiaq Qadri 《World Journal of Gastroenterology》 SCIE CAS 2014年第35期12462-12472,共11页
Chronic active hepatitis(CAH) is acknowledged as an imperative risk factor for the development of liver injury and hepatocellular carcinoma.The histological end points of CAH are chronic inflammation,fibrosis and cirr... Chronic active hepatitis(CAH) is acknowledged as an imperative risk factor for the development of liver injury and hepatocellular carcinoma.The histological end points of CAH are chronic inflammation,fibrosis and cirrhosis which are coupled with increased DNA synthesis in cirrhotic vs healthy normal livers.The potential mechanism involved in CAH includes a combination of processes leading to liver cell necrosis,inflammation and cytokine production and liver scaring(fibrosis).The severity of liver damage is regulated by Hepatitis B virus genotypes and viral components.The viral and cellular factors that contribute to liver injury are discussed in this article.Liver injury caused by the viral infection affects many cellular processes such as cell signaling,apoptosis,transcription,DNA repair which in turn induce radical effects on cell survival,growth,transformation and maintenance.The consequence of such perturbations is resulted in the alteration of bile secretion,gluconeogenesis,glycolysis,detoxification and metabolism of carbohydrates,proteins,fat and balance of nutrients.The identification and elucidation of the molecular pathways perturbed by the viral proteins are important in order to design effective strategy to minimize and/or restore the hepatocytes injury. 展开更多
关键词 hepatitis B virus hepatitis B virus genotype Hepatocellular carcinoma woodchuck hepatitis virus Ground squirrel hepatitis virus Peripheral blood mononuclear cells Interferon regulatory factor 7 Interleukin-1 receptor-associated kinase 4 TNF receptor-associated factor 3
下载PDF
New therapeutic vaccination strategies for the treatment of chronic hepatitis B 被引量:9
7
作者 Jia Liu Anna Kosinska +1 位作者 Mengji Lu Michael Roggendorf 《Virologica Sinica》 SCIE CAS CSCD 2014年第1期10-16,共7页
Chronic hepatitis B virus(CHB) is currently treated with either interferon-based or nucleot(s)idebased antiviral therapies.However,treatment with pegylated interferon alpha results in a durable antiviral response in o... Chronic hepatitis B virus(CHB) is currently treated with either interferon-based or nucleot(s)idebased antiviral therapies.However,treatment with pegylated interferon alpha results in a durable antiviral response in only about 30%patients and is associated with side effects.Most patients receiving nucleot(s)ide analogue treatment do not establish long-term,durable control of Infection and have rebounding viremia after cessation of therapy.Thus,novel therapy strategies are necessary to achieve the induction of potent and durable antiviral immune responses of the patients which can maintain long-term control of viral replication.Therapeutic vaccination of HBV carriers is a promising strategy for the control of hepatitis B.Here the authors review new therapeutic vaccination strategies to treat chronic hepatitis B which may be introduced for patient treatment in the future. 展开更多
关键词 hepatitis B virus woodchuck hepatitis virus therapeutic vaccination IMMUNOMODULATION programmed death-1
下载PDF
Recent Advances in Research on Hepadnaviral Infection in the Woodchuck Model
8
作者 Ina Schulte E-juan ZHANG +3 位作者 Zhong-ji MENG Rong-juan PEI Mengji LU Michael Roggendorf 《Virologica Sinica》 SCIE CAS CSCD 2008年第2期107-115,共9页
The woodchuck model is an excellent animal model to study hepadnaviral infection. The new progresses in this model made possible to examine the T-cell mediated immune responses in acute and chronic hepadnaviral infect... The woodchuck model is an excellent animal model to study hepadnaviral infection. The new progresses in this model made possible to examine the T-cell mediated immune responses in acute and chronic hepadnaviral infection. Recently, a new assay for cytotoxic T-cells based on detection of CD107 was established for the woodchuck model. In addition, new immunotherapeutic approaches based on combination of potent antiviral treatment and DNA-protein vaccines were proven to be useful for treatment of chronic hepatitis B. 展开更多
关键词 Chronic hepatitis B woodchuck hepatitis virus whv hepatitis B virus (HBV)
下载PDF
Molecular Cloning,Characterization and Expression Analysis of Woodchuck Retinoic Acid-Inducible GeneⅠ
9
作者 严琦 刘钦 +11 位作者 李蒙蒙 李芳慧 朱彬 Jun-zhong WANG 王俊忠 卢银平 刘嘉 吴珺 郑昕 陆蒙吉 王宝菊 杨东亮 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2016年第3期335-343,共9页
Cytosolic retinoic acid-inducible gene I(RIG-I) is an important innate immune RNA sensor and can induce antiviral cytokines, e.g., interferon-β(IFN-β). Innate immune response to hepatitis B virus(HBV) plays a ... Cytosolic retinoic acid-inducible gene I(RIG-I) is an important innate immune RNA sensor and can induce antiviral cytokines, e.g., interferon-β(IFN-β). Innate immune response to hepatitis B virus(HBV) plays a pivotal role in viral clearance and persistence. However, knowledge of the role that RIG-I plays in HBV infection is limited. The woodchuck is a valuable model for studying HBV infection. To characterize the molecular basis of woodchuck RIG-I(w RIG-I), we analyzed the complete coding sequences(CDSs) of w RIG-I, containing 2778 base pairs that encode 925 amino acids. The deduced w RIG-I protein was 106.847 k D with a theoretical isoelectric point(p I) of 6.07, and contained three important functional structures [caspase activation and recruitment domains(CARDs), DEx D/H-box helicases, and a repressor domain(RD)]. In woodchuck fibroblastoma cell line(WH12/6), w RIG-I-targeted small interfering RNA(si RNA) down-regulated RIG-I and its downstrean effector–IFN-β transcripts under RIG-I' ligand, 5'-ppp double stranded RNA(ds RNA) stimulation. We also measured m RNA levels of w RIG-I in different tissues from healthy woodchucks and in the livers from woodchuck hepatitis virus(WHV)-infected woodchucks. The basal expression levels of w RIG-I were abundant in the kidney and liver. Importantly, w RIG-I was significantly up-regulated in acutely infected woodchuck livers, suggesting that RIG-I might be involved in WHV infection. These results may characterize RIG-I in the woodchuck model, providing a strong basis for further study on RIG-I-mediated innate immunity in HBV infection. 展开更多
关键词 retinoic acid-inducible gene I woodchuck woodchuck hepatitis virus
下载PDF
Asymptomatic Hepadnaviral Persistence and Its Consequences in the Woodchuck Model of Occult Hepatitis B Virus Infection 被引量:7
10
作者 Patricia M.Mulrooney-Cousins Tomasz I.Michalak 《Journal of Clinical and Translational Hepatology》 SCIE 2015年第3期211-219,共9页
Woodchuck hepatitis virus (WHV) is molecularly and pathogenically closely related to hepatitis B virus (HBV).Both viruses display tropism towards hepatocytes and cells of the immune system and cause similar liver path... Woodchuck hepatitis virus (WHV) is molecularly and pathogenically closely related to hepatitis B virus (HBV).Both viruses display tropism towards hepatocytes and cells of the immune system and cause similar liver pathology,where acute hepatitis can progress to chronic hepatitis and to hepatocellular carcinoma (HCC).Two forms of occult hepadnaviral persistence were identified in the woodchuck-WHV model:secondary occult infection (SOI) and primary occult infection (POI).SOI occurs after resolution of a serologically apparent infection with hepatitis or after subclinical serologically evident virus exposure.POI is caused by small amounts of virus and progresses without serological infection markers,but the virus genome and its replication are detectable in the immune system and with time in the liver.SOI can be accompanied by minimal hepatitis,while the hallmark of POI is normal liver morphology.Nonetheless,HCC develops in about 20% of animals with SOI or POI within 3 to 5 years.The virus persists throughout the lifespan in both SOI and POI at serum levels rarely greater than 100 copies/mL,causes hepatitis and HCC when concentrated and administered to virus-na(i)ve woodchucks.SOI is accompanied by virusspecific T and B cell immune responses,while only virusspecific T cells are detected in POI.SOI coincides with protection against reinfection,while POI does not and hepatitis develops after challenge with liver pathogenic doses >1000 virions.Both SOI and POI are associated with virus DNA integration into the liver and the immune system genomes.Overall,SOI and POI are two distinct forms of silent hepadnaviral persistence that share common characteristics.Here,we review findings from the woodchuck model and discuss the relevant observations made in human occult HBV infection (OBI). 展开更多
关键词 hepatitis B Occult hepatitis B virus infection woodchuck model of hepatitis B woodchuck hepatitis virus Secondary occult infection Primary occult infection Consequences of occult hepadnaviral infection Hepatocellular carcinoma
原文传递
中国青海地区喜马拉雅旱獭嗜肝病毒自然感染的组织学研究 被引量:14
11
作者 李新宇 王宝菊 +4 位作者 孟忠吉 汪由坤 赵西平 陆蒙吉 杨东亮 《中国病毒学》 CAS CSCD 2005年第6期570-573,共4页
应用原位杂交技术、免疫组化技术以土拨鼠肝炎病毒(Woodchuck hepatitis virus,WHV)的检测系统检测50份 喜马拉雅旱獭肝组织可能存在的嗜肝病毒c基因、s抗原及c抗原的表达,同时检测肝脏组织病理学改变。结果显示 旱獭肝组织中嗜肝病毒s... 应用原位杂交技术、免疫组化技术以土拨鼠肝炎病毒(Woodchuck hepatitis virus,WHV)的检测系统检测50份 喜马拉雅旱獭肝组织可能存在的嗜肝病毒c基因、s抗原及c抗原的表达,同时检测肝脏组织病理学改变。结果显示 旱獭肝组织中嗜肝病毒s抗原、c抗原的阳性率分别为26%(13/50)、36%(18/50);在抗原双阳性的10份肝组织标本 中有c基因的阳性表达,阳性率为50%。c抗原定位于肝细胞胞浆和/或胞核,呈散在、片簇状分布,c基因定位于肝细 胞的细胞核,阳性细胞散在分布。50份标本中5份出现肝炎的病理改变,与抗原检出间无明显相关性。使用WHV 的病毒检测系统证实青海地区喜马拉雅旱獭可能存在类似WHV的嗜肝病毒感染,从组织学的角度为中国青海地区 喜马拉雅旱獭嗜肝病毒自然感染提供证据,此种动物有可能用于建立嗜肝病毒感染的动物模型。 展开更多
关键词 青海喜马拉雅旱獭 正嗜肝 DNA 病毒属 土拔鼠肝炎病毒 原位杂交 免疫组化
下载PDF
丁型肝炎病毒感染东方土拔鼠的实验研究 被引量:4
12
作者 金志宏 杨波 +9 位作者 杨小昂 赵国龙 焉晋询 马丽丽 汲振余 陈洪涛 路浩 李艳军 孟林敏 买凯 《病毒学报》 CAS CSCD 北大核心 1990年第1期74-76,共3页
丁型肝炎病毒(HDV)原称Delta因子,是一种缺陷性RNA病毒。它的复制需依赖于乙型肝炎病毒(HBV)的辅助作用。由于HBV的体外细胞培养很难成功,故HDV的研究仅限于动物模型。Rizzetto等首先对黑猩猩进行HDV的实验感染并获成功。Summers等在东... 丁型肝炎病毒(HDV)原称Delta因子,是一种缺陷性RNA病毒。它的复制需依赖于乙型肝炎病毒(HBV)的辅助作用。由于HBV的体外细胞培养很难成功,故HDV的研究仅限于动物模型。Rizzetto等首先对黑猩猩进行HDV的实验感染并获成功。Summers等在东方土拨鼠(Marmota monax)中发现一种病毒,性质与HBV相似,称为土拨鼠肝炎病毒(Woodchuck Hepatitis Virus,WHV)。WHV可使土拨鼠患病毒性肝炎和肝细胞癌。Ponzetto等用慢性携带WHV的土拔鼠进行了HDV感染研究,发现WHV对HDV同样有辅助作用,故土拔鼠可作为研究人类肝炎和肝细胞癌的动物模型。目前,国内尚无HDV实验感染的报告。本文对HDV感染东方土拨鼠进行了实验研究,试图探讨土拔鼠丁型肝炎血清学及病理学改变,为提纯丁型肝炎病毒及其抗原提供实验材料。 展开更多
关键词 丁型肝炎病毒 东方土拔鼠 感染
下载PDF
Toll样受体激动剂抑制土拨鼠肝炎病毒作用研究 被引量:5
13
作者 孟忠吉 张永红 +4 位作者 李文兵 汤守兵 李东 柯昌征 陈悦 《肝脏》 2013年第6期378-381,共4页
目的研究Toll样受体(TLR)激动剂对土拨鼠肝炎病毒(WHV)复制的抑制作用。方法从WHV慢性感染的土拨鼠肝脏分离原代肝细胞,转染Poly I:C、CpG或直接用LPS刺激,Southern印迹检测WHV复制中间体,病毒保护试验检测细胞培养上清液中I型干扰素的... 目的研究Toll样受体(TLR)激动剂对土拨鼠肝炎病毒(WHV)复制的抑制作用。方法从WHV慢性感染的土拨鼠肝脏分离原代肝细胞,转染Poly I:C、CpG或直接用LPS刺激,Southern印迹检测WHV复制中间体,病毒保护试验检测细胞培养上清液中I型干扰素的水平;通过尾静脉注射Poly I:C、LPS、CpG到WHV转基因小鼠体内,提取肝脏DNA和RNA,实时PCR检测MxA和OAS1的mRNA水平。结果 Poly I:C、LPS和CpG在原代土拨鼠肝细胞和WHV转基因小鼠显著抑制病毒复制;Poly I:C、LPS和CpG处理后的原代肝细胞分泌高水平的I型干扰素,肝脏的MxA和OAS1 mRNA上调表达。结论TLR通路的活化通过诱导I型干扰素而发挥抗病毒作用,在抗HBV治疗中可能发挥重要作用。 展开更多
关键词 TOLL样受体 土拨鼠肝炎病毒 原代肝细胞 转基因小鼠
下载PDF
土拨鼠肝炎病毒核心蛋白质粒的构建、原核表达及多克隆抗体的制备与鉴定 被引量:2
14
作者 张振华 田拥军 +4 位作者 李磊 王宝菊 孟忠吉 陆蒙吉 杨东亮 《华中科技大学学报(医学版)》 CAS CSCD 北大核心 2007年第5期567-570,577,F0002,共6页
目的构建土拨鼠肝炎病毒核心蛋白质粒并进行原核表达、抗体制备。方法应用基因工程技术将编码截短型土拨鼠肝炎病毒核心蛋白(WHcAg1~149aa)的基因片段装入原核表达载体pQE60上,在JM109菌内进行诱导表达,使用切胶回收及Ni-NTA柱两种方... 目的构建土拨鼠肝炎病毒核心蛋白质粒并进行原核表达、抗体制备。方法应用基因工程技术将编码截短型土拨鼠肝炎病毒核心蛋白(WHcAg1~149aa)的基因片段装入原核表达载体pQE60上,在JM109菌内进行诱导表达,使用切胶回收及Ni-NTA柱两种方法纯化目的蛋白。将纯化蛋白免疫家兔制备多克隆抗体,并采用酶联免疫吸附实验、免疫组织化学及Western blot检测抗体的灵敏度和特异性。结果成功地构建了含截短型土拨鼠肝炎病毒核心区基因的质粒并获得表达,经纯化得到了分子量约为16.5kD的重组核心蛋白,免疫家兔获得了高效价的特异性多克隆抗体,而且与HBcAg有交叉反应。结论获得的重组截短型土拨鼠肝炎病毒核心抗原(1~149aa)纯度高,免疫原性强。获得的兔抗-WHc效价高,特异性好,与HBcAg有交叉反应。 展开更多
关键词 土拨鼠肝炎病毒 核心蛋白 原核表达 多克隆抗体 抗原抗体交叉反应
下载PDF
RNA干扰抑制土拨鼠肝炎病毒X基因表达 被引量:4
15
作者 孟忠吉 吴珺 +4 位作者 张永红 汤守兵 李东 柯昌征 陈悦 《湖北医药学院学报》 CAS 2011年第5期475-479,共5页
目的:研究土拨鼠肝炎病毒(WHV)序列特异性siRNA对WHV X基因表达的抑制作用。方法:以WHV全长质粒为模板PCR扩增WHV X基因,克隆到pXF3H载体构建WHX-HA融合蛋白真核表达质粒(pXF3H-WHx);合成siRNA的模板序列,克隆到siRNA表达质粒psiRNA构建... 目的:研究土拨鼠肝炎病毒(WHV)序列特异性siRNA对WHV X基因表达的抑制作用。方法:以WHV全长质粒为模板PCR扩增WHV X基因,克隆到pXF3H载体构建WHX-HA融合蛋白真核表达质粒(pXF3H-WHx);合成siRNA的模板序列,克隆到siRNA表达质粒psiRNA构建shRNA表达质粒(psiWx1、psiWx2);质粒pXF3H-WHx与psiWx1、psiWx2共转染Hela细胞,Western blot检测WHV X蛋白的表达。结果:Hela细胞中WHVX蛋白的表达受siRNA的抑制,psiWx1表达的siRNA抑制90%以上的X蛋白表达,并呈剂量依赖性,但是WHV X基因靶位点中2个突变的核苷酸可使这种干扰作用消失。结论:质粒表达的siRNA序列特异性抑制WHV X蛋白的表达。 展开更多
关键词 土拨鼠肝炎病毒 土拨鼠肝炎病毒X蛋白 RNA干扰 小干扰RNA
下载PDF
土拨鼠肝炎病毒转基因小鼠模型的建立 被引量:1
16
作者 孟忠吉 张永红 +3 位作者 柯昌征 李东 汤守兵 陈悦 《湖北医药学院学报》 CAS 2014年第4期321-326,F0002,共7页
目的:建立土拨鼠肝炎病毒(Woodchuck hepatitis virus,WHV)转基因小鼠模型,以用于嗜肝病毒感染的发病机制研究。方法:采用微注射技术将1.3-倍WHV基因[野生型和WHV S抗原(WHV surface antigen,WHsAg)缺陷型]注射入C57BL/6xC3H小鼠... 目的:建立土拨鼠肝炎病毒(Woodchuck hepatitis virus,WHV)转基因小鼠模型,以用于嗜肝病毒感染的发病机制研究。方法:采用微注射技术将1.3-倍WHV基因[野生型和WHV S抗原(WHV surface antigen,WHsAg)缺陷型]注射入C57BL/6xC3H小鼠的胚胎,然后与C57BL/6小鼠回交,建立WHV转基因小鼠。肝内WHV DNA复制和mRNA表达水平分别采用Southern杂交和real-time RT-PCR检测,血清中WHV病毒载量采用real-time PCR检测,HE染色进行肝组织病理学检测,ELISA检测血清中WHV核心抗体,流式细胞仪检测WHV核心抗原(WHV core antigen,WHcAg)特异性细胞毒性T淋巴细胞。结果:成功建立野生型WHV转基因小鼠和WHsAg缺陷型转基因小鼠。WHsAg缺陷型小鼠肝内WHV复制水平显著高于野生型小鼠,野生型WHV转基因小鼠血清WHV载量104-108copies/mL。而且雄性小鼠肝内和血清WHV复制水平显著高于雌性小鼠。16周龄以上的WHV转基因小鼠约半数血清可检测出高滴度的WHV核心抗体。而且少数转基因小鼠可以检测到WHcAg特异性细胞毒性T淋巴细胞。结论:这种新建的WHV转基因小鼠肝内可检测到病毒复制和mRNA表达,血清有高水平的WHV病毒载量,可以用于嗜肝病毒感染的发病机制、抗病毒治疗等研究。 展开更多
关键词 土拨鼠肝炎病毒 转基因小鼠 复制 细胞免疫
下载PDF
ELISA检测土拨鼠肝炎病毒核心抗体方法的建立 被引量:1
17
作者 鲍俊杰 王宝菊 +3 位作者 田拥军 江敏 陆蒙吉 杨东亮 《肝脏》 2007年第3期183-185,共3页
目的建立检测土拨鼠肝炎病毒(WHV)核心抗体的ELISA方法,应用于WHV感染的中国旱獭动物模型的研究。方法原核表达重组WHcAg,氯化铯密度梯度离心获得非变性的纯化蛋白;用该纯化蛋白免疫Balb/c小鼠制备多克隆抗体;建立竞争抑制ELISA方法用... 目的建立检测土拨鼠肝炎病毒(WHV)核心抗体的ELISA方法,应用于WHV感染的中国旱獭动物模型的研究。方法原核表达重组WHcAg,氯化铯密度梯度离心获得非变性的纯化蛋白;用该纯化蛋白免疫Balb/c小鼠制备多克隆抗体;建立竞争抑制ELISA方法用于检测旱獭血清中的抗-WHc。结果纯化后的蛋白浓度达0.86mg/mL,纯度达89.48%;免疫小鼠后获得抗血清,ELISA间接法显示其多克隆抗体效价达1∶640000,Western blot分析显示该抗体能特异识别WHcAg;建立的竞争抑制ELISA方法对旱獭血清中可能存在的抗-WHc进行检测,诊断特异性及敏感性均较好,重复测定的批内变异系数和批间变异系数均小于10%。结论成功地建立检测旱獭血清中抗-WHc的竞争抑制ELISA方法。该方法具有稳定、简便、特异、敏感的特点,适用于大量旱獭血清抗-WHc的筛查工作,为进一步研究中国旱獭这一新型乙型肝炎病毒动物模型奠定了基础。 展开更多
关键词 土拨鼠肝炎病毒核心抗体 ELISA 中国旱獭
下载PDF
RNA干扰上调天然免疫 被引量:1
18
作者 孟忠吉 张永红 +3 位作者 李东 汤守兵 柯昌征 陈悦 《湖北医药学院学报》 CAS 2013年第4期281-285,共5页
目的:研究特异性RNA干扰对于天然免疫的上调作用。方法:分别分离WHV感染的土拨鼠和WHV转基因小鼠原代肝细胞,转染针对WHV、小鼠β-actin和GAPDH的siRNA。Northern blot和Real-time RT-PCR分别检测原代肝细胞中WHV、MxA、MHCⅠ、β-actin... 目的:研究特异性RNA干扰对于天然免疫的上调作用。方法:分别分离WHV感染的土拨鼠和WHV转基因小鼠原代肝细胞,转染针对WHV、小鼠β-actin和GAPDH的siRNA。Northern blot和Real-time RT-PCR分别检测原代肝细胞中WHV、MxA、MHCⅠ、β-actin、GAPDH、IFN-β和IP-10的mRNA水平。结果:针对WHV、小鼠β-actin和GAPDH的siRNA抑制靶基因mRNA伴随MxA、MHCⅠ、IP-10等干扰素刺激基因的上调表达。这种干扰素刺激基因的上调表达只有在靶基因被特异siRNA降解时出现,呈剂量和沉寂效应依赖性,而且可以被siRNA特异性抑制剂所阻断。结论:RNAi上调天然免疫可能增强siRNA的抗病毒效应,有可能在抗病毒RNAi研究中发挥重要作用。 展开更多
关键词 RNA干扰 土拨鼠肝炎病毒 原代肝细胞 天然免疫 干扰素刺激基因
下载PDF
siRNA联合恩替卡韦在原代肝细胞中抑制土拨鼠肝炎病毒基因表达和复制 被引量:1
19
作者 孟忠吉 张永红 +3 位作者 柯昌征 李东 汤守兵 陈悦 《湖北医药学院学报》 CAS 2013年第1期35-38,43,共5页
目的:在原代土拨鼠肝细胞中研究siRNA联合恩替卡韦(ETV)对土拨鼠肝炎病毒(WHV)基因表达和复制的抑制作用。方法:分别设计合成针对WHV PreS1、S、C、X基因的4种siRNA,转染原代土拨鼠肝细胞,同时给予ETV处理,Southern和Northern印迹检测WH... 目的:在原代土拨鼠肝细胞中研究siRNA联合恩替卡韦(ETV)对土拨鼠肝炎病毒(WHV)基因表达和复制的抑制作用。方法:分别设计合成针对WHV PreS1、S、C、X基因的4种siRNA,转染原代土拨鼠肝细胞,同时给予ETV处理,Southern和Northern印迹检测WHV病毒复制中间体和mRNA水平,realtime PCR检测细胞上清中WHV DNA。结果:4种siRNA对WHV基因表达和复制的作用显示出不同的动力学特征,siWHs和siWHx的抑制作用最强,而siWHpreS1和siWHc的活性相对较弱。在原代肝细胞中,首先出现的是WHV mRNA水平的降低,然后是WHV复制中间体水平的降低,siWHs和siWHx最高抑制60%~70%的mRNA和病毒复制中间体,同时抑制细胞上清中90%的WHV DNA。另外siWHx与ETV联合应用不仅可以增强抗病毒作用,而且可以防止ETV停药反跳。结论:siRNA在WHV自然感染的原代肝细胞中可以有效抑制病毒的基因表达和复制,因而siRNA有可能开发成为新的抗HBV药物,尤其适用于核苷(酸)类似物的联合应用。 展开更多
关键词 RNA干扰 小干扰性RNA 土拨鼠肝炎病毒 原代肝细胞 核苷(酸)类似物
下载PDF
抗土拨鼠肝炎病毒核心蛋白单克隆抗体的制备、性质鉴定及初步应用
20
作者 张振华 田拥军 +4 位作者 李磊 张珺 龚劲松 陆蒙吉 杨东亮 《中国免疫学杂志》 CAS CSCD 北大核心 2007年第4期341-344,共4页
目的研制出能特异与土拨鼠肝炎病毒核心蛋白(WHc)结合的单克隆抗体,使之能特异性地对土拨鼠肝炎病毒(WHV)进行检测,并可能应用于相关肝炎病毒的筛查。方法以原核表达的土拨鼠肝炎病毒重组核心蛋白(WHc 1~149氨基酸)免疫BALB/c小... 目的研制出能特异与土拨鼠肝炎病毒核心蛋白(WHc)结合的单克隆抗体,使之能特异性地对土拨鼠肝炎病毒(WHV)进行检测,并可能应用于相关肝炎病毒的筛查。方法以原核表达的土拨鼠肝炎病毒重组核心蛋白(WHc 1~149氨基酸)免疫BALB/c小鼠,常规杂交瘤技术进行细胞融合,有限稀释法克隆化,间接酶联免疫吸附试验(ELISA)和免疫组织化学(IHC)筛选、鉴定。结果筛选出5株(4B1E、6C5D、6C5C、6D1D、6D1G)能稳定分泌抗土拨鼠肝炎病毒核心蛋白抗体的杂交瘤细胞株。此5株单抗适用于ELISA、IHC、Western blot等方面的研究,与HBcAg有交叉反应,并且在部分中国旱獭的肝脏组织进行IHC检测呈现阳性反应。结论制备的5株单抗可用于土拨鼠肝炎病毒等嗜肝脱氧核糖核酸病毒的研究,可能在寻找新的相关肝炎病毒中起重要作用。 展开更多
关键词 土拨鼠肝炎病毒 核心蛋白 单克隆抗体 旱獭
下载PDF
上一页 1 2 下一页 到第
使用帮助 返回顶部