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Establishment of an ELISA to Detect Kaposi's Sarcoma-associated Herpesvirus Using Recombinant ORF73 被引量:8
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作者 Xin-xing OUYANG Bi-shi FU +3 位作者 Bao-lin LI Yan ZENG Fan-hong XU Lin-ding WANG 《Virologica Sinica》 SCIE CAS CSCD 2010年第3期168-176,共9页
Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 p... Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Han people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005). 展开更多
关键词 基因蛋白 疱疹病毒 ELISA检测 重组 肉瘤 ELISA法 PAGE电泳 因果关系
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Expression of Kaposi's Sarcoma-associated Herpesvirus ORFK8.1 and Its Preliminary Diagnostic Application 被引量:6
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作者 Bi-shi FU Bao-lin LI +3 位作者 Xin-xing OUYANG Yan ZENG Fan-hong XU Lin-ding WANG 《Virologica Sinica》 SCIE CAS CSCD 2009年第3期202-208,共7页
The ORFK8.1 of Kaposi's sarcoma associated-herpesvirus(KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogala... The ORFK8.1 of Kaposi's sarcoma associated-herpesvirus(KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogalactopyranoside(IPTG) . The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis(SDS-PAGE) . SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant ORFK8.1 protein. The optimal condition of the recombinant ORFK8.1 ELISA assay was confirmed:the concentration of antigen was 5 μg/mL,the dilution of serum was 1:200. We used the ELISA method to investigate the recombinant ORF K8.1 protein's specificity,the data showed that the specificity of ORF K8.1 to detect KSHV was 100%. At the same time,560 sera samples from Hubei province were detected by using ORFK8.1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%. 展开更多
关键词 疱疹病毒 肉瘤 ELISA检测 重组大肠杆菌 聚丙烯酰胺凝胶电泳 ELISA法 十二烷基硫酸钠 蛋白纯化
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The Biology of Kaposi’s Sarcoma-Associated Herpesvirus and the Infection of Human Immunodeficiency Virus 被引量:1
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作者 Di QIN Chun LU 《Virologica Sinica》 SCIE CAS CSCD 2008年第6期473-485,共13页
Kaposi sarcoma-associated herpesvirus (KSHV),also known as human herpesvirus 8 (HHV-8),is discovered in 1994 from Kaposi's sarcoma (KS) lesion of an acquired immunodeficiency syndrome (AIDS) patient. In addition t... Kaposi sarcoma-associated herpesvirus (KSHV),also known as human herpesvirus 8 (HHV-8),is discovered in 1994 from Kaposi's sarcoma (KS) lesion of an acquired immunodeficiency syndrome (AIDS) patient. In addition to its association with KS,KSHV has also been implicated as the causative agent of two other AIDS-associated malignancies: primary effusion lymphoma (PEL) and multicentric Castleman’s disease (MCD). KSHV is a complex DNA virus that not only has the ability to promote cellular growth and survival for tumor development,but also can provoke deregulated angiogenesis,inflammation,and modulate the patient’s immune system in favor of tumor growth. As KSHV is a necessary but not sufficient etiological factor for KS,human immunodeficiency virus (HIV) is a very important cofactor. Here we review the basic information about the biology of KSHV,development of pathogenesis and interaction between KSHV and HIV. 展开更多
关键词 生物学 卡波西氏肉瘤 疱疹病毒 免疫缺陷 临床分析
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Pathogenesis and Associated Diseases of Kaposi's Sarcoma-associated Herpesvirus 被引量:2
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作者 Lin-ding WANG 《中国病毒学》 CSCD 2007年第3期248-255,共8页
Kaposi's sarcoma-associated herpesvirus(KSHV)is the primary etiological agent of Kaposi's sarcoma,primary effusion lymphoma and muticentric Castleman's disease.In common with the other herpesviruses,KSHV e... Kaposi's sarcoma-associated herpesvirus(KSHV)is the primary etiological agent of Kaposi's sarcoma,primary effusion lymphoma and muticentric Castleman's disease.In common with the other herpesviruses,KSHV exhibits both latent and lytic life cycles,both of which are characterized by distinct gene expression profiles and programs.KSHV encodes proteins which play essential roles in the inhibition of host adaptive and innate immunity,the inhibition of apoptosis,and the regulation of the cell cycle.KSHV also encodes several proteins which have transforming and intrcellular signalling activity. 展开更多
关键词 发病机理 疱疹病毒 卡波济氏肉瘤 多中心性巨大淋巴结增生病
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Angiogenesis,Kaposi’s Sarcoma and Kaposi’s Sarcoma-associated Herpesvirus
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作者 Tao KANG Feng-chun Ye +1 位作者 Shou-jiang gao Lin-ding WAN 《Virologica Sinica》 SCIE CAS CSCD 2008年第6期449-458,共10页
Tumor angiogenesis is the uncontrolled growth of blood vessels in tumors,serving to supply nutrients and oxygen,and remove metabolic wastes. Kaposi’s sarcoma (KS),a multifocal angioproliferative disorder characterize... Tumor angiogenesis is the uncontrolled growth of blood vessels in tumors,serving to supply nutrients and oxygen,and remove metabolic wastes. Kaposi’s sarcoma (KS),a multifocal angioproliferative disorder characterized by spindle cell proliferation,neo-angiogenesis,inflammation,and edema,is associated with infection by Kaposi's sarcoma-associated herpesvirus (KSHV). Recent studies indicate that KSHV infection directly promotes angiogenesis and inflammation through an autocrine and paracrine mechanism by inducing pro-angiogenic and pro-inflammatory cytokines. Many of these cytokines are also expressed in KS lesions,implicating a direct role of KSHV in the pathogenesis of this malignancy. Several KSHV genes are involved in KSHV-induced angiogenesis. These studies have provided insights into the pathogenesis of KS,and identified potential therapeutic targets for this malignancy. 展开更多
关键词 血管 卡波西氏肉瘤 疱疹病毒 临床分析
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Immunogenicity Analysis of Prokaryotic Expression Products of Kaposi's Sarcoma Associated Herpesvirus orf65 被引量:6
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作者 Bi-shi FU Bao-lin LI Lin-ding WANG 《Virologica Sinica》 SCIE CAS CSCD 2008年第3期196-202,共7页
To purify the protein encoding the small capsid protein (SCP) of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi's sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic... To purify the protein encoding the small capsid protein (SCP) of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi's sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA. 展开更多
关键词 免疫性分析 kaposiS肉瘤 疱疹病毒 治疗方法
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维吾尔族Kaposi’s肉瘤患者组织中KSHV-vFLIP表达水平与患者临床特征相关性 被引量:3
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作者 王鹏 冯燕艳 +1 位作者 张泽高 马云霞 《中国皮肤性病学杂志》 CAS CSCD 北大核心 2016年第11期1120-1122,1191,共4页
目的探讨维吾尔族卡波西肉瘤(Kaposi’s sarcoma,KS)患者皮损组织内KSHV-vFLIP基因表达水平与临床特征的相关性。方法选取29例维吾尔族卡波西肉瘤患者和12例血管瘤(hemangioma)患者、11例健康对照患者皮损石蜡包埋组织,反转录-聚合酶链... 目的探讨维吾尔族卡波西肉瘤(Kaposi’s sarcoma,KS)患者皮损组织内KSHV-vFLIP基因表达水平与临床特征的相关性。方法选取29例维吾尔族卡波西肉瘤患者和12例血管瘤(hemangioma)患者、11例健康对照患者皮损石蜡包埋组织,反转录-聚合酶链反应(RT-PCR)分析KSHV-vFLIP基因mRNA表达水平。结果 KS患者KSHV-vFLIP基因表达水平(4.436±0.508)显著高于健康对照组(1.145±0.112,P<0.05),其中斑块型组KSHV-vFLIP基因表达水平(8.104±2.225)显著高于结节型组(3.971±0.371,P<0.05)和斑片组(2.774±0.867,P<0.05)。而KS组(4.436±0.507)与血管瘤组(6.343±0.949)差异无统计学意义(P>0.05),且HIV携带组(3.739±0.816)与非HIV携带组(4.618±0.604)差异无统计学意义(P>0.05)。结论在维吾尔族卡波西肉瘤患者组织中,KSHV-vFLIP基因表达水平与瘤体体积正相关。 展开更多
关键词 卡波西肉瘤 kshv-vFLIP基因 实时荧光定量 石蜡包埋 瘤体体积
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一种新的人类疱疹病毒:Kaposi’s肉瘤相关疱疹病毒(KSHV)
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作者 王世珍 刘淑红 耿运琪 《中国病毒学》 CAS CSCD 1999年第1期1-6,共6页
1994年Y.Chang等人从AIDS患者的Kaposi’s肉瘤(KS)组织中分离到了两种独有的序列[1],并由KS建立了基因组文库,对其中外源序列进行克隆和序列分析[2],初步确认一种新的人类疱疹病毒———KS相关... 1994年Y.Chang等人从AIDS患者的Kaposi’s肉瘤(KS)组织中分离到了两种独有的序列[1],并由KS建立了基因组文库,对其中外源序列进行克隆和序列分析[2],初步确认一种新的人类疱疹病毒———KS相关疱疹病毒。1KSHV的发现Kapo... 展开更多
关键词 Kapsi's肉瘤 艾滋病 8型 人类疱疹病毒
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Regulation of Wnt/β-catenin signaling by herpesviruses 被引量:2
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作者 Kevin J Zwezdaryk Joseph A Combs +1 位作者 Cindy A Morris Deborah E Sullivan 《World Journal of Virology》 2016年第4期144-154,共11页
The Wnt/β-catenin signaling pathway is instrumental in successful differentiation and proliferation of mammalian cells. It is therefore not surprising that the herpesvirus family has developed mechanisms to interact ... The Wnt/β-catenin signaling pathway is instrumental in successful differentiation and proliferation of mammalian cells. It is therefore not surprising that the herpesvirus family has developed mechanisms to interact with and manipulate this pathway. Successful coexistence with the host requires that herpesviruses establish a lifelong infection that includes periods of latency and reactivation or persistence. Many herpesviruses establish latency in progenitor cells and viral reactivation is linked to host-cell proliferation and differentiation status. Importantly, Wnt/β-catenin is tightly connected to stem/progenitor cell maintenance and differentiation. Numerous studies have linked Wnt/β-catenin signaling to a variety of cancers, emphasizing the importance of Wnt/β-catenin pathways in development, tissue homeostasis and disease. This review details how the alpha-, beta-, and gammaherpesviruses interact and manipulate the Wnt/β-catenin pathway to promote a virus-centric agenda. 展开更多
关键词 herpesvirus Herpes simplex virus-1 VARICELLA ZOSTER VIRUS Cytomegalovirus Epstein-Barr VIRUS kaposi’s sarcoma-associated herpesvirus WNT/Β-CATENIN Glycogen synthase kinase-3 Axin
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The Role of Human Herpesvirus 8 Molecular Characterization in the Management of HIV Infected Patients Diagnosed with Malignancies Associated with Its Infection
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作者 Martínez Pedro Ariel Kourí Vivian +11 位作者 Blanco Orestes Capó Virginia Abad Yoandra Alemán Yoan Verdasquera Denis Jiménez Narciso Caballero Iraida Fleites Gilberto Ugarte Yaumara Calderón Odalys álvarez Alina Ulrich Hengge 《World Journal of AIDS》 2013年第3期221-230,共10页
Despite the progress has been reached with Human herpesvirus 8 (HHV-8) research, there are gaps in the knowledge of viral induced oncogenesis. The aim of the present study was to identify possible associations between... Despite the progress has been reached with Human herpesvirus 8 (HHV-8) research, there are gaps in the knowledge of viral induced oncogenesis. The aim of the present study was to identify possible associations between HHV-8 subtypes, HHV-8 loads and clinical manifestations of HIV infected patients diagnosed with different malignancies associated with HHV-8 infection. Forty six HIV-1 infected individuals diagnosed with different HHV-8 associated diseases were studied [37 epidemic Kaposi’s sarcoma (KS), 3 pleural effusion lymphoma (PEL);5 peripheral lymphadenopathies (PL);1 Hodgkin’s lymphoma (HL);1 non Hodgkin’s lymphoma (NHL)]. HHV-8 loads were determined by quantitative real time PCR (qRT-PCR) whilst HHV-8 subtypes were determined by open-reading frame (ORF)-K1 gen genotyping. HHV-8 subtypes B, A, C, A5 and E were exhibited by 31.8%, 23.4%, 19.1%, 17% and 8.5% of the studied patients, respectively. The median HHV-8 viral load did not differ between subtypes (p > 0.05) but HHV-8 viral loads were significantly higher in PEL than in epidemic KS lesion or lymph nodes (p = 0.04). Subtype B was detected in 60% of patients with B cell lymphoma (NHL, PEL and HL) whereas subtype E was only detected in patients with epidemic KS diagnosis. Our data suggest that HHV-8 DNA quantification instead of subtype identification could be used as a surrogate marker for monitoring its infection, not only in epidemic KS patients but also in HIV infected individuals with lymphoproliferative disorders. 展开更多
关键词 Human herpesvirus 8 or kaposi’s sarcoma-associated herpesvirus Real Time PCR SUBTYPES LYMPHOPROLIFERATIVE Disorders Cuban HIV/AIDS
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Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis 被引量:3
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作者 Denis Avey +3 位作者 Brittany Brewers Fanxiu Zhu 《Virologica Sinica》 SCIE CAS CSCD 2015年第2期130-145,共16页
It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epid... It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis. 展开更多
关键词 kaposi’s sarcoma-associated herpesvirus(kshv) kaposi’s SARCOMA epigenetic EPIGENOME herpesvirus oncovirus chromatin
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Prevalence of Kaposi's sarcoma-associated herpesvirus in Uygur and Han populations from the Urumqi and Kashgar regions of Xinjiang, China 被引量:5
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作者 Jun Zheng Yang Yang +6 位作者 Meng Cui Zhan-Jun Shu Li-Li Han Zhen-Qiu Liu Charles Wood Tiejun Zhang Yan Zeng 《Virologica Sinica》 CAS CSCD 2017年第5期396-403,共8页
Kaposi's sarcoma-associated herpesvirus(KSHV) is the infectious etiologic agent associated with Kaposi's sarcoma(KS), primary effusion lymphoma, and multicentric Castleman disease. It has been shown that high ... Kaposi's sarcoma-associated herpesvirus(KSHV) is the infectious etiologic agent associated with Kaposi's sarcoma(KS), primary effusion lymphoma, and multicentric Castleman disease. It has been shown that high KSHV prevalence and high incidence of both classic KS and AIDSassociated KS are found mostly among people of Uygur ethnicity in Xinjiang, while people of Han ethnicity in Xinjiang have a higher KSHV seroprevalence than those of other Han populations in China's Mainland. However, it is still unclear why there is such geographical and population variation in KSHV distribution in China. In this work, we focused on the populations in the Kashgar region and Urumqi area, where a total of 1294 research subjects were randomly selected to investigate the potential correlation between KSHV prevalence and different ethnicities in endemic areas of Xinjiang, and to determine risk factors that may affect KSHV infection rates or KS incidence. We identified a high seroprevalence of KSHV and high peripheral blood DNA infection in the general Uygur and Han populations in both Urumqi and Kashgar regions of Xinjiang, and determined that advancing age, low education level, and stationary population status affect KSHV infection rates. Further, KSHV-positive Uygur participants were shown to have higher prevalence of neutralizing antibodies and neutralizing antibody titers than KSHV-positive Han participants. 展开更多
关键词 kaposi’s sarcoma associated herpesvirus(kshv) PREVALENCE Uygur ethnicity Han ethnicity XINJIANG
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Lipids, lipid metabolism and Kaposi's sarcoma-associated herpesvirus pathogenesis
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作者 Lu Dai Zhen Lin +2 位作者 Wei Jiang Erik K.Flemington Zhiqiang Qin 《Virologica Sinica》 SCIE CAS CSCD 2017年第5期369-375,共7页
Lipids are essential for mammalian cells to maintain many physiological functions. Emerging evidence has shown that cancer cells can develop specific alterations in lipid biosynthesis and metabolism to facilitate thei... Lipids are essential for mammalian cells to maintain many physiological functions. Emerging evidence has shown that cancer cells can develop specific alterations in lipid biosynthesis and metabolism to facilitate their survival and various malignant behaviors. To date, the precise role of cellular lipids and lipid metabolism in viral oncogenesis is still largely unclear with only a handful of literature covering this topic to implicate lipid metabolism in oncogenic virus associated pathogenesis. In this review, we focus on the role of lipid biosynthesis and metabolism in the pathogenesis of the Kaposi's sarcoma-associated herpesvirus, a common causative factor for cancers arising in the immunocompromised settings. 展开更多
关键词 kaposi’s sarcoma-associated herpesvirus(kshv) herpesvirus lipid metabolism
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Prevalence of Kaposi's sarcoma-associated herpesvirus among intravenous drug users: a systematic review and meta-analysis
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作者 Qiwen Fang Zhenqiu Liu +2 位作者 Zhijie Zhang Yan Zeng Tiejun Zhang 《Virologica Sinica》 SCIE CAS CSCD 2017年第5期415-422,共8页
Intravenous drug users(IDUs) have been demonstrated to be highly vulnerable to HIV/AIDS.Nevertheless, the prevalence of Kaposi's sarcoma associated herpesvirus(KSHV), an important co-infected agent with HIV, among... Intravenous drug users(IDUs) have been demonstrated to be highly vulnerable to HIV/AIDS.Nevertheless, the prevalence of Kaposi's sarcoma associated herpesvirus(KSHV), an important co-infected agent with HIV, among this population remained obscure. We conducted a systematic review on the epidemiological features of KSHV among IDUs worldwide. Eligible studies were retrieved from 6 electronic databases(Pub Med, EMBASE, Web of Science, CBM, CNKI and Wanfang).We calculated the pooled prevalence and 95% confidence interval(CI) overall and among subgroups using either random-effects model or fixed-effects model depending on between-study heterogeneity. The potential publication bias was assessed by the Egger's test. A meta-regression analysis was performed to explore the sources of heterogeneity. Finally, twenty-two studies with a total sample of 7881 IDUs were included in the analysis. The pooled prevalence of KSHV was14.71%(95% CI 11.12%–19.46%) among IDUs. Specifically, KSHV prevalence was 10.86%(95% CI6.95%–16.96%) in HIV-negative IDUs, and 13.56%(95% CI 10.57%–17.38%) in HIV-positive IDUs.Moreover, prevalence among IDUs from the three continents involved in the current study was similar:16.10%(95%CI 7.73%–33.54%) in Asia; 14.22%(95%CI 8.96%–22.57%) in Europe and 14.06%(95%CI11.38%–17.37%) in America. Globally, IDUs are at higher risk of the KSHV infection when compared with the general population, regardless of geographical region or HIV-infection status. 展开更多
关键词 kaposi’s sarcoma-associated herpesvirus(kshv) PREVALENCE intravenous drug users(IDUs)
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Allicin and Glycyrrhizic Acid Display Antiviral Activity Against Latent and Lytic Kaposi Sarcoma-associated Herpesvirus 被引量:1
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作者 Yirui Xie Ying Chen +2 位作者 Yongzheng Guo Ying Huang Biao Zhu 《Infectious Microbes & Diseases》 2020年第1期30-34,共5页
Kaposi sarcoma-associated herpesvirus(KSHV)triggers the development of Kaposi sarcoma,a skin malignancy that is one of the most widespread defining symptoms in acquired immunodeficiency syndrome patients.KSHV manifest... Kaposi sarcoma-associated herpesvirus(KSHV)triggers the development of Kaposi sarcoma,a skin malignancy that is one of the most widespread defining symptoms in acquired immunodeficiency syndrome patients.KSHV manifests in two distinct cycles,a chronic latent cycle and an acute lytic cycle.Current clinical anti-herpesvirus therapeutic agents are predominantly composed of nucleoside analogues that target viral replication in the lytic cycle only,while KSHV latent genes are at the basis tumorigenesis.Currently,there are no effective therapies targeting latent KSHV infections.Therefore,the aim of this study was to identify putative therapeutic compounds with inhibitory activity against latent KSHV.The KSHV-infected primary effusion lymphoma cell line BC-3 was used to study antiviral activity of glycyrrhizic acid(GA),Allicin,and epigallocatechin-3-gallate(EGCG)against latent and lytic KSHV.Activity of GA,Allicin,EGCG,and the established anti-lytic cycle control compound ganciclovir was quantified by real-time polymerase chain reaction of nuclear and virion KSHV DNA yields after treatment compared with the untreated control.GA and Allicin showed antiviral activity against both latent and lytic KSHV,while EGCG displayed activity against lytic KSHV only.Therefore,GA and Allicin are interesting compounds for further development of anti-KSHV therapy against latent cycle infections. 展开更多
关键词 kaposi sarcoma-associated herpesvirus BC-3 cells ALLICIN glycyrrhizic acid
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The replication and transcription activator (RTA) of Kaposi's sarcoma-associated herpesvirus/human herpesvirus-8
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作者 Zhilong YANG Charles WOOD 《Frontiers in Biology》 CSCD 2010年第2期105-115,共11页
Kaposi’s sarcoma-associated herpesvirus(KSHV)is γ-2 herpesvirus with latency and lytic replication stages in its life-cycle.The viral replication and transcription activator(RTA)is the key protein for triggering KSH... Kaposi’s sarcoma-associated herpesvirus(KSHV)is γ-2 herpesvirus with latency and lytic replication stages in its life-cycle.The viral replication and transcription activator(RTA)is the key protein for triggering KSHV lytic gene expression and replication from latency.In this review,we will discuss the gene expression program in KSHV lytic replication and latency,the regulation of the RTA expression,the RTA protein and the mechanisms that RTA utilizes to transactivate its target genes.We will focus on the RTA-mediated transactivation mechanisms,including DNA-binding,interacting with cellular co-factors and promoting repressor degradation. 展开更多
关键词 kaposi’s sarcoma-associated herpesvirus(kshv) replication and transcription activator(RTA) TRANSACTIVATION
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KSHV ORF73C基因原核表达产物的纯化与免疫原性分析 被引量:1
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作者 汤巧 卢春 +3 位作者 黄丽 钱超 曾怡 张玲 《临床检验杂志》 CAS CSCD 北大核心 2004年第5期364-366,共3页
目的 纯化卡波济肉瘤相关疱疹病毒 (KSHV)潜伏相关核抗原 (LANA)羧基端编码基因ORF73C原核表达蛋白并分析其免疫原性。方法 含重组质粒pGEX 6p 1+ORF73C(pORF73C)的宿主菌BL2 1经异丙基硫代 β D 半乳糖苷 (IPTG)诱导表达。用glutath... 目的 纯化卡波济肉瘤相关疱疹病毒 (KSHV)潜伏相关核抗原 (LANA)羧基端编码基因ORF73C原核表达蛋白并分析其免疫原性。方法 含重组质粒pGEX 6p 1+ORF73C(pORF73C)的宿主菌BL2 1经异丙基硫代 β D 半乳糖苷 (IPTG)诱导表达。用glutathioneSepharose 4B亲和层析柱纯化表达产物后 ,以重组 3C蛋白酶对融合蛋白进行解离 ,SDS PAGE对表达及纯化产物进行鉴定。以纯化的ORF73C蛋白免疫小鼠获得抗血清 ,建立ELISA检测其免疫原性。结果 SDS PAGE显示蛋白条带大小分别为 4 90 0 0和 2 30 0 0 ,与预期的ORF73C融合蛋白及ORF73C蛋白分子量大小一致 ;ELISA检测显示 ,抗ORF73C多抗免疫反应性高于未免疫小鼠血清 2倍以上。结论 成功获得了纯化的ORF73C蛋白 ,经ELISA显示其有较强免疫原性。 展开更多
关键词 C蛋白 免疫原性 纯化 原核表达产物 C基因 免疫小鼠 核抗原 IPTG 诱导表达 SDS-PAGE
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HHV6感染激活卡波济肉瘤相关疱疹病毒(KSHV)的溶解性周期复制 被引量:1
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作者 曾怡 卢春 《病毒学报》 CAS CSCD 北大核心 2005年第2期93-100,共8页
运用细胞融合、细胞混合培养、条件培养基培养和病毒直接刺激等方法,研究人类疱疹病毒6型(HHV6)对卡波济肉瘤相关疱疹病毒(KSHV)溶解性周期复制的影响。①将HHV6感染的JJhan细胞(T淋巴细胞系)与BCBL-1细胞(原发性渗出性淋巴瘤,PEL)进行... 运用细胞融合、细胞混合培养、条件培养基培养和病毒直接刺激等方法,研究人类疱疹病毒6型(HHV6)对卡波济肉瘤相关疱疹病毒(KSHV)溶解性周期复制的影响。①将HHV6感染的JJhan细胞(T淋巴细胞系)与BCBL-1细胞(原发性渗出性淋巴瘤,PEL)进行细胞融合形成异核体细胞。②将HHV6感染的JJhan细胞与BCBL-1细胞进行混合培养。③收集HHV6感染的JJhan细胞培养上清液作为条件培养基进行灭活处理,以灭活前后的条件培养基培养BCBL-1细胞。进一步离心纯化HHV6病毒颗粒,并感染BCBL-1细胞,分别设紫外线和热灭活的HHV6病毒颗粒感染BCBL-1细胞为对照。提取上述的实验细胞总RNA,RT-PCR和/或实时定量(Real—time)PCR检测卡波济肉瘤相关疱疹病毒(KSHV)次要衣壳蛋白编码基因ORF26mRNA转录。结果显示:①细胞融合后15h开始出现明显细胞病变,RT-PCR检测不同时间的实验组ORF26mRNA转录水平均明显高于对照组;Real—timePCR检测各时间ORF26mRNA转录水平是对照组的2.3倍以上;②细胞混合培养72h时,实验组ORF26mRNA转录水平是对照组的1.8倍;混合培养5天时,实验组KSHV裂解周期蛋白K8.1表达水平是对照组的2.46倍;③灭活前后的HHV6感染细胞培养上清液培养BCBL-1细胞96h时,ORF26mRNA转录水平分别是对照组的2.73倍和2.22倍;④灭活前后的HHV6均可增强BCBL-1细胞中KSHVORF26mRNA转录水平。提示:HHV6感染可激活KSHV的溶解性周期复制。 展开更多
关键词 感染 对照组 细胞 卡波济肉瘤 MRNA转录 灭活 疱疹病毒 ORF2 PCR检测 性周期
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含KSHV vIL-6基因原核表达载体的构建及融合蛋白的表达纯化与鉴定 被引量:1
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作者 葛高霞 王平 卢春 《江苏大学学报(医学版)》 CAS 2010年第3期244-247,共4页
目的:在大肠埃希菌中表达卡波济肉瘤相关疱疹病毒(Kaposi′s sarcoma-associated herpesvirus,KSHV)vIL-6基因,并进一步纯化和鉴定表达蛋白。方法:以本实验室已构建的重组质粒pGEX-6p-1-vIL-6为模板,PCR扩增目的基因片段,将PCR产物克隆... 目的:在大肠埃希菌中表达卡波济肉瘤相关疱疹病毒(Kaposi′s sarcoma-associated herpesvirus,KSHV)vIL-6基因,并进一步纯化和鉴定表达蛋白。方法:以本实验室已构建的重组质粒pGEX-6p-1-vIL-6为模板,PCR扩增目的基因片段,将PCR产物克隆至原核表达载体pET-32a(+)中,构建vIL-6的重组原核表达质粒pET-32a(+)-vIL-6。将重组质粒转化大肠埃希菌(E.coli)BL21(DE3)感受态细胞,用异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达融合蛋白。表达产物通过镍亲和层析柱纯化并经蛋白质印迹法鉴定。结果:限制性内切酶检测和基因测序证实,成功构建了含有vIL-6基因的原核表达载体。蛋白质印迹法结果显示,在大肠埃希菌BL21(DE3)中His-Tag融合蛋白得到了表达,亲和层析法纯化后获得了相对分子质量为43 000的vIL-6融合蛋白。结论:重组质粒pET-32a(+)-vIL-6能在大肠埃希菌BL21(DE3)中表达,利用镍亲和层析柱可纯化获得融合蛋白。 展开更多
关键词 病毒白细胞介素6基因 原核表达 融合蛋白 卡波济肉瘤相关疱疹病毒
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新疆某地维吾尔族男性吸毒者KSHV血清流行病学初步分析
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作者 杨培荣 郭淑霞 +5 位作者 谭晓华 杨磊 付碧石 王林定 张景玉 张国宁 《石河子大学学报(自然科学版)》 CAS 2010年第1期68-71,共4页
为进一步研究HIV-KSHV感染相关疾病提供基础资料,利用ELISA法检测KSHV潜伏期LANA1、溶解期ORF65、K8.1抗原抗体,采用χ2检验分析有关数据,分析了新疆某地维吾尔族男性吸毒人群的KSHV血清流行病学的特点。吸毒人群主要以注射吸毒者为主(1... 为进一步研究HIV-KSHV感染相关疾病提供基础资料,利用ELISA法检测KSHV潜伏期LANA1、溶解期ORF65、K8.1抗原抗体,采用χ2检验分析有关数据,分析了新疆某地维吾尔族男性吸毒人群的KSHV血清流行病学的特点。吸毒人群主要以注射吸毒者为主(184/199,92.5%),在199份吸毒者血清中共检测到KSHV阳性血清60份,KSHV血清阳性率是30.6%;χ2检验分析显示,年龄、职业、文化程度、婚姻状况、HIV-1血清状态、吸毒方式以及吸毒年限均与KSHV血清阳性率无关,该吸毒人群中KSHV感染率较高,表明KSHV普遍存在于新疆某地维族男性吸毒人群。 展开更多
关键词 卡波氏肉瘤相关病疱疹毒 吸毒人群 血清流行病学
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