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Virus-Like Particles Harboring CCL19, IL-2 and HPV16 E7 Elicit Protective T Cell Responses in HLA-A2 Transgenic Mice. 携带CCL19、IL-2和hpv16e7的病毒样颗粒在HLA-A2转基因小鼠中引发保护性T细胞反应
Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1874357901206010270
Victoria Juarez, H Amalia Pasolli, Andrea Hellwig, Natalio Garbi, Angel Cid Arregui

Infection by high-risk genotypes of human papillomaviruses (HR-HPVs) is the cause of cancer of the uterine cervix. Although prophylactic vaccines directed against the two most prevalent HR-HPV types (HPV16 and 18) have been commercialized recently, there is a need for effective therapeutic vaccines against HR-HPVs. We have tested in mice a chimeric protein composed of the hepatitis B small surface antigen (HBsAg(S)) flanked at its N-terminus by chemokine CC ligand 19/macrophage inflammatory protein-3β (CCL19/MIP-3β), and at the C-terminus by interleukin 2 (IL-2) and an artificial HPV16 E7 polytope. This protein is assembled into nanoparticles and both CCL19 and IL-2 conserve their functionality. HLA-A2 (AAD) transgenic mice immunized with a plasmid encoding this protein mounted specific T cell responses against E7 without the need of an adjuvant. Furthermore, vaccination prevented the development of tumors after implantation of the E6/E7-expressing TC-1/A2 tumor cell line. Our results suggest that vaccines based on HBsAg(S) nanoparticles carrying short E7 epitopes and immune-stimulatory domains might be of therapeutic value in the treatment of patients suffering from cervical pre-cancer or cancer lesions caused by HR-HPVs.

高危基因型人乳头瘤病毒(hr - hpv)感染是子宫颈癌的原因。尽管针对两种最流行的人乳头瘤病毒类型(HPV16和18)的预防性疫苗最近已经商业化,但仍需要针对人乳头瘤病毒的有效治疗性疫苗。我们在小鼠中测试了一种由乙型肝炎小表面抗原(HBsAg(S))组成的嵌合蛋白,该嵌合蛋白在其n端由趋化因子CC配体19/巨噬细胞炎症蛋白3β (CCL19/MIP-3β)组成,在其c端由白细胞介素2 (IL-2)和人工hpv16e7多肽组成。这种蛋白质被组装成纳米颗粒,CCL19和IL-2都保留了它们的功能。用编码该蛋白的质粒免疫的HLA-A2 (AAD)转基因小鼠在不需要佐剂的情况下对E7产生特异性T细胞反应。此外,在植入表达E6/ e7的TC-1/A2肿瘤细胞系后,接种疫苗可以阻止肿瘤的发展。我们的研究结果表明,基于携带短E7表位和免疫刺激结构域的HBsAg(S)纳米颗粒的疫苗可能在治疗由hr - hpv引起的宫颈癌前病变或癌症病变的患者中具有治疗价值。
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引用次数: 20
The HPV E2-Host Protein-Protein Interactions: A Complex Hijacking of the Cellular Network. HPV e2 -宿主蛋白-蛋白相互作用:细胞网络的复杂劫持。
Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1874357901206010173
Mandy Muller, Caroline Demeret

Over 100 genotypes of human papillomaviruses (HPVs) have been identified as being responsible for unapparent infections or for lesions ranging from benign skin or genital warts to cancer. The pathogenesis of HPV results from complex relationships between viral and host factors, driven in particular by the interplay between the host proteome and the early viral proteins. The E2 protein regulates the transcription, the replication as well as the mitotic segregation of the viral genome through the recruitment of host cell factors to the HPV regulatory region. It is thereby a pivotal factor for the productive viral life cycle and for viral persistence, a major risk factor for cancer development. In addition, the E2 proteins have been shown to engage numerous interactions through which they play important roles in modulating the host cell. Such E2 activities are probably contributing to create cell conditions appropriate for the successive stages of the viral life cycle, and some of these activities have been demonstrated only for the oncogenic high-risk HPV. The recent mapping of E2-host protein-protein interactions with 12 genotypes representative of HPV diversity has shed some light on the large complexity of the host cell hijacking and on its diversity according to viral genotypes. This article reviews the functions of E2 as they emerge from the E2/host proteome interplay, taking into account the large-scale comparative interactomic study.

人类乳头瘤病毒(hpv)的100多种基因型已被确定为导致隐性感染或从良性皮肤或生殖器疣到癌症等病变的原因。HPV的发病机制是由病毒和宿主因素之间的复杂关系引起的,特别是由宿主蛋白质组和早期病毒蛋白之间的相互作用驱动的。E2蛋白通过将宿主细胞因子募集到HPV调控区,调控病毒基因组的转录、复制以及有丝分裂分离。因此,它是多产的病毒生命周期和病毒持久性的关键因素,是癌症发展的主要危险因素。此外,E2蛋白已被证明参与许多相互作用,通过它们在调节宿主细胞中发挥重要作用。这种E2活性可能有助于为病毒生命周期的连续阶段创造适合的细胞条件,其中一些活性仅在致癌高风险的HPV中得到证实。最近绘制的e2 -宿主蛋白-蛋白质相互作用与HPV多样性的12个基因型的代表,揭示了宿主细胞劫持的复杂性及其根据病毒基因型的多样性。本文回顾了E2与宿主蛋白质组相互作用中E2的功能,并考虑了大规模的比较相互作用研究。
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引用次数: 0
RNA interference for the treatment of papillomavirus disease. RNA干扰治疗乳头瘤病毒病。
Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1874357901206010204
Richa Singhania, Norliana Khairuddin, Daniel Clarke, Nigel Aj McMillan

Human Papillomavirus (HPV)-induced diseases are a significant burden on our healthcare system and current therapies are not curative. Vaccination provides significant prophylactic protection but effective therapeutic treatments will still be required. RNA interference (RNAi) has great promise in providing highly specific therapies for all HPV diseases yet this promise has not been realised. Here we review the research into RNAi therapy for HPV in vitro and in vivo and examine the various targets and outcomes. We discuss the idea of using RNAi with current treatments and address delivery of RNAi, the major issue holding back clinical adoption. Finally, we present our view of a potential path to the clinic.

人乳头瘤病毒(HPV)引起的疾病是我们医疗保健系统的一个重大负担,目前的治疗方法无法治愈。疫苗接种提供了重要的预防性保护,但仍需要有效的治疗方法。RNA干扰(RNAi)在为所有HPV疾病提供高度特异性治疗方面有很大的希望,但这一希望尚未实现。本文综述了体外和体内RNAi治疗HPV的研究进展,并探讨了各种靶点和结果。我们讨论了在当前治疗中使用RNAi的想法,并解决了RNAi的递送问题,这是阻碍临床采用的主要问题。最后,我们提出了我们的观点,一个潜在的路径到诊所。
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引用次数: 0
Human papillomavirus infections and cancer stem cells of tumors from the uterine cervix. 人乳头瘤病毒感染与子宫颈肿瘤的癌干细胞。
Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1874357901206010232
Jacqueline López, Graciela Ruíz, Jorge Organista-Nava, Patricio Gariglio, Alejandro García-Carrancá

Different rate of development of productive infections (as low grade cervical intraepithelial neoplasias), or high grade lesions and cervical malignant tumors associated with infections of the Transformation zone (TZ) by High-Risk Human Papillomavirus (HR-HPV), could suggest that different epithelial host target cells could exist. If there is more than one target cell, their differential infection by HR-HPV may play a central role in the development of cervical cancer. Recently, the concept that cancer might arise from a rare population of cells with stem cell-like properties has received support in several solid tumors, including cervical cancer (CC). According to the cancer stem cell (CSC) hypothesis, CC can now be considered a disease in which stem cells of the TZ are converted to cervical cancer stem cells by the interplay between HR-HPV viral oncogenes and cellular alterations that are thought to be finally responsible for tumor initiation and maintenance. Current studies of CSC could provide novel insights regarding tumor initiation and progression, their relation with viral proteins and interplay with the tumor micro-environment. This review will focus on the biology of cervical cancer stem cells, which might contribute to our understanding of the mechanisms responsible for cervical tumor development.

与高危人乳头瘤病毒(HR-HPV)感染转化区(TZ)相关的生产性感染(如低级别宫颈上皮内瘤变)或高级别病变和宫颈恶性肿瘤的发展速度不同,可能表明存在不同的上皮宿主靶细胞。如果存在不止一种靶细胞,那么它们被 HR-HPV 感染的不同程度可能在宫颈癌的发展过程中起到核心作用。最近,包括宫颈癌(CC)在内的几种实体瘤都支持癌症可能源自具有干细胞特性的稀有细胞群这一概念。根据癌症干细胞(CSC)假说,宫颈癌现在可以被认为是一种疾病,在这种疾病中,TZ 干细胞在 HR-HPV 病毒致癌基因和细胞改变的相互作用下转化为宫颈癌干细胞,而细胞改变被认为是肿瘤发生和维持的最终原因。目前对宫颈癌干细胞的研究可以提供有关肿瘤发生和发展、其与病毒蛋白的关系以及与肿瘤微环境相互作用的新见解。本综述将重点讨论宫颈癌干细胞的生物学特性,这可能有助于我们了解宫颈肿瘤的发生机制。
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引用次数: 0
Laboratory detection of respiratory viruses by automated techniques. 利用自动化技术对呼吸道病毒进行实验室检测。
Pub Date : 2012-01-01 Epub Date: 2012-11-30 DOI: 10.2174/1874357901206010151
Mercedes Pérez-Ruiz, Irene Pedrosa-Corral, Sara Sanbonmatsu-Gámez, María Navarro-Marí

Advances in clinical virology for detecting respiratory viruses have been focused on nucleic acids amplification techniques, which have converted in the reference method for the diagnosis of acute respiratory infections of viral aetiology. Improvements of current commercial molecular assays to reduce hands-on-time rely on two strategies, a stepwise automation (semi-automation) and the complete automation of the whole procedure. Contributions to the former strategy have been the use of automated nucleic acids extractors, multiplex PCR, real-time PCR and/or DNA arrays for detection of amplicons. Commercial fully-automated molecular systems are now available for the detection of respiratory viruses. Some of them could convert in point-of-care methods substituting antigen tests for detection of respiratory syncytial virus and influenza A and B viruses. This article describes laboratory methods for detection of respiratory viruses. A cost-effective and rational diagnostic algorithm is proposed, considering technical aspects of the available assays, infrastructure possibilities of each laboratory and clinic-epidemiologic factors of the infection.

临床病毒学在检测呼吸道病毒方面的进展主要集中在核酸扩增技术上,该技术已成为诊断急性呼吸道病毒感染的参考方法。目前商业分子检测方法的改进主要依靠两种策略来减少操作时间:逐步自动化(半自动化)和整个过程完全自动化。自动核酸提取器、多重 PCR、实时 PCR 和/或用于检测扩增子的 DNA 阵列为前一种策略做出了贡献。目前已有用于检测呼吸道病毒的商用全自动分子系统。其中一些系统可转化为护理点方法,取代抗原检测法,用于检测呼吸道合胞病毒、甲型和乙型流感病毒。本文介绍了实验室检测呼吸道病毒的方法。考虑到现有检测方法的技术方面、各实验室基础设施的可能性以及感染的临床流行病学因素,提出了一种具有成本效益的合理诊断算法。
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引用次数: 0
Improving Clinical Laboratory Efficiency: Introduction of Systems for the Diagnosis and Monitoring of HIV Infection. 提高临床实验室效率:引进HIV感染诊断和监测系统。
Pub Date : 2012-01-01 Epub Date: 2012-11-30 DOI: 10.2174/1874357901206010135
Marta Alvarez, Natalia Chueca, Vicente Guillot, María Del Carmen Bernal, Federico García

Since the first tests for identifying individuals with suspected human immunodeficiency virus (HIV) infection were introduced in the mid-1980s, diagnostic virology testing has greatly evolved. The technological advances, automating in the laboratories and the advances in molecular biology techniques have helped introduce invaluable laboratory methods for managing HIV patients. Tests for diagnosis, specially for screening HIV antibodies, are now fully automated; in the same way, tests for monitoring HIV viral load (HIV RNA copies/ml of plasma), which is used for monitoring infection and response to antiretroviral treatment, are also fully automated; however, resistance testing, tropism determination and minor variant detection, which are used to make decisions for changing antiretroviral treatment regimens in patients failing therapy, still remain highly laborious and time consuming. This chapter will review the main aspects relating to the automating of the methods available for laboratory diagnosis as well as for monitoring of the HIV infection and determination of resistance to antiretrovirals and viral tropism.

自20世纪80年代中期首次引入用于识别疑似人类免疫缺陷病毒(HIV)感染个体的测试以来,诊断病毒学测试已经有了很大的发展。技术进步、实验室自动化和分子生物学技术的进步有助于引入宝贵的实验室方法来管理艾滋病毒患者。诊断测试,特别是筛选艾滋病毒抗体的测试,现在已经完全自动化;同样,用于监测感染和对抗逆转录病毒治疗的反应的监测艾滋病毒载量(血浆中艾滋病毒RNA拷贝数/毫升)的测试也是完全自动化的;然而,用于在治疗失败的患者中决定改变抗逆转录病毒治疗方案的耐药检测、倾向测定和微小变异检测仍然非常费力和耗时。本章将回顾与实验室诊断方法自动化有关的主要方面,以及监测艾滋病毒感染和确定对抗逆转录病毒药物和病毒趋向性的耐药性。
{"title":"Improving Clinical Laboratory Efficiency: Introduction of Systems for the Diagnosis and Monitoring of HIV Infection.","authors":"Marta Alvarez,&nbsp;Natalia Chueca,&nbsp;Vicente Guillot,&nbsp;María Del Carmen Bernal,&nbsp;Federico García","doi":"10.2174/1874357901206010135","DOIUrl":"https://doi.org/10.2174/1874357901206010135","url":null,"abstract":"<p><p>Since the first tests for identifying individuals with suspected human immunodeficiency virus (HIV) infection were introduced in the mid-1980s, diagnostic virology testing has greatly evolved. The technological advances, automating in the laboratories and the advances in molecular biology techniques have helped introduce invaluable laboratory methods for managing HIV patients. Tests for diagnosis, specially for screening HIV antibodies, are now fully automated; in the same way, tests for monitoring HIV viral load (HIV RNA copies/ml of plasma), which is used for monitoring infection and response to antiretroviral treatment, are also fully automated; however, resistance testing, tropism determination and minor variant detection, which are used to make decisions for changing antiretroviral treatment regimens in patients failing therapy, still remain highly laborious and time consuming. This chapter will review the main aspects relating to the automating of the methods available for laboratory diagnosis as well as for monitoring of the HIV infection and determination of resistance to antiretrovirals and viral tropism.</p>","PeriodicalId":23111,"journal":{"name":"The Open Virology Journal","volume":"6 ","pages":"135-43"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2174/1874357901206010135","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31144494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 10
Immunotherapy of human papilloma virus induced disease. 人乳头瘤病毒诱导疾病的免疫治疗。
Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1874357901206010257
Sjoerd H van der Burg

Immunotherapy is the generic name for treatment modalities aiming to reinforce the immune system against diseases in which the immune system plays a role. The design of an optimal immunotherapeutic treatment against chronic viruses and associated diseases requires a detailed understanding of the interactions between the target virus and its host, in order to define the specific strategies that may have the best chance to deliver success at each stage of disease. Recently, a first series of successes was reported for the immunotherapy of Human Papilloma Virus (HPV)-induced premalignant diseases but there is definitely room for improvement. Here I discuss a number of topics that in my opinion require more study as the answers to these questions allows us to better understand the underlying mechanisms of disease and as such to tailor treatment.

免疫疗法是旨在加强免疫系统对抗免疫系统起作用的疾病的治疗方式的总称。设计针对慢性病毒和相关疾病的最佳免疫治疗方法需要详细了解目标病毒与其宿主之间的相互作用,以便确定在疾病的每个阶段最有可能取得成功的具体策略。最近,免疫治疗人类乳头状瘤病毒(HPV)诱导的恶性前病变取得了一系列成功,但肯定还有改进的余地。在这里,我讨论了一些我认为需要更多研究的主题,因为这些问题的答案使我们能够更好地理解疾病的潜在机制,从而制定治疗方案。
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引用次数: 9
Genome Stability of Pandemic Influenza A (H1N1) 2009 Based on Analysis of Hemagglutinin and Neuraminidase Genes. 基于血凝素和神经氨酸酶基因分析的2009年甲型H1N1流感基因组稳定性
Pub Date : 2012-01-01 Epub Date: 2012-04-26 DOI: 10.2174/1874357901206010059
Emilio E Espínola

Influenza A virus (H1N1), which arose in 2009, constituted the fourth pandemic after the cases of 1918, 1957, and 1968. This new variant was formed by a triple reassortment, with genomic segments from swine, avian, and human influenza origins. The objective of this study was to analyze sequences of hemagglutinin (n=2038) and neuraminidase (n=1273) genes, in order to assess the extent of diversity among circulating 2009-2010 strains, estimate if these genes evolved through positive, negative, or neutral selection models of evolution during the pandemic phase, and analyze the worldwide percentage of detection of important amino acid mutations that could enhance the viral performance, such as transmissibility or resistance to drugs. A continuous surveillance by public health authorities will be critical to monitor the appearance of new influenza variants, especially in animal reservoirs such as swine and birds, in order to prevent the potential animal-human transmission of viruses with pandemic potential.

2009年出现的甲型流感病毒(H1N1)是继1918年、1957年和1968年之后的第四次大流行。这种新变种是由猪流感、禽流感和人类流感的基因组片段三重重组形成的。本研究的目的是分析血球凝集素(n=2038)和神经氨酸酶(n=1273)基因序列,以评估2009-2010年流行毒株之间的多样性程度,估计这些基因在大流行阶段是通过阳性、阴性还是中性的进化选择模型进化,并分析全球范围内检测到的能够增强病毒性能(如传播性或耐药性)的重要氨基酸突变的百分比。公共卫生当局的持续监测对于监测新的流感变体的出现至关重要,特别是在猪和鸟类等动物宿主中,以防止具有大流行潜力的病毒在动物-人类之间的潜在传播。
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引用次数: 10
The E89K Mutation in the Matrix Protein of the Measles Virus Affects In Vitro Cell Death and Virus Replication Efficiency in Human PBMC. 麻疹病毒基质蛋白E89K突变影响人PBMC体外细胞死亡和病毒复制效率
Pub Date : 2012-01-01 Epub Date: 2012-06-08 DOI: 10.2174/1874357901206010068
Jianbao Dong, Wei Zhu, Akatsuki Saito, Yoshitaka Goto, Hiroyuki Iwata, Takeshi Haga

Matrix protein is known to have an important role in the process of virus assembly and virion release during measles virus replication. In the present in vitro study, a single mutation of E89K in the matrix protein was shown to affect cell death and virus replication efficiency in human PBMC. One strain with this mutation caused less cell death than the parental virus, and possessed high virus replication efficiency. Moreover, by Annexin V-FITC staining, polycaspase FLICA staining, and double labeling with poly-caspase FLICA and the Hoechst stain, the cell death seen was shown to be apoptosis.

在麻疹病毒复制过程中,基质蛋白在病毒组装和病毒粒子释放过程中起着重要作用。在目前的体外研究中,基质蛋白中E89K的单个突变被证明可以影响人PBMC细胞死亡和病毒复制效率。有这种突变的一个毒株比亲本病毒造成的细胞死亡少,并且具有很高的病毒复制效率。Annexin V-FITC染色、polycaspase FLICA染色、polycaspase FLICA和Hoechst染色双标记显示细胞死亡为凋亡。
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引用次数: 2
Development and Field Testing of a Real-Time PCR Assay for Caprine Arthritis-Encephalitis-Virus (CAEV). 山羊关节炎-脑炎病毒(CAEV)实时荧光定量PCR检测方法的建立及现场试验
Pub Date : 2012-01-01 Epub Date: 2011-07-27 DOI: 10.2174/1874357901206010082
Giovanni Brajon, Daniela Mandas, Manuele Liciardi, Flavia Taccori, Mauro Meloni, Franco Corrias, Caterina Montaldo, Ferdinando Coghe, Cristina Casciari, Monica Giammarioli, Germano Orrù

Caprine arthritis/encephalitis (CAE) of goats and occasionally sheep are persistent virus infections caused by a lentivirus (CAEV). This viral infection results in arthritis in adult animals and encephalitis in kids. Prognosis for the encephalitic form is normally poor, with substantial economic loss for the farm. In this context an early/fast laboratory diagnosis for CAEV infection could be useful for effective prophylactic action. In this work we performed a quantitative real time PCR designed on the CAEV env gene to detect/quantify in goat/sheep samples, viral RNA or proviral DNA forms of CAEV. This procedure was validated in 15 sheep, experimentally infected with CAEV or with a highly correlated lentivirus (visna maedi, MVV); in addition, a total of 37 clinical goat specimens recruited in CAEV positive herds were analyzed and compared using serological analysis (Elisa and AGID). All samples infected with MVV resulted negative. In sheep experimentally infected with CAEV, proviral DNA was detectable 15 days post infection, whereas the serological methods revealed an indicative positivity after 40-60 days.This method showed a sensitivity of 10(2) env fragments/PCR) with a linear dynamic range of quantitation from 10(3) to 10(7)env fragments/PCR; the R2 correlation coefficient was 0.98. All subjects with a clinical diagnosis for Caprine Arthritis-Encephalitis (CAE) resulted CAEV DNA positive.

山羊和绵羊的山羊关节炎/脑炎(CAE)是由慢病毒(CAEV)引起的持续性病毒感染。这种病毒感染导致成年动物关节炎和儿童脑炎。脑病形式的预后通常很差,给养殖场造成重大经济损失。在这种情况下,CAEV感染的早期/快速实验室诊断可能有助于采取有效的预防措施。在这项工作中,我们对CAEV的env基因进行了定量实时PCR设计,以检测/定量山羊/绵羊样品中CAEV的病毒RNA或前病毒DNA形式。该方法在15只羊身上得到了验证,这些羊被实验性地感染了CAEV或高度相关的慢病毒(maedi visna, MVV);此外,采用血清学分析(Elisa和AGID)对CAEV阳性牧群中37份临床山羊标本进行分析和比较。所有感染MVV的样本结果均为阴性。在实验感染CAEV的绵羊中,感染15天后可检测到前病毒DNA,而血清学方法在40-60天后显示指示性阳性。该方法灵敏度为10(2)个env片段/PCR,定量线性动态范围为10(3)~ 10(7)个env片段/PCR;R2相关系数为0.98。所有临床诊断为山羊关节炎-脑炎(CAE)的受试者均为CAEV DNA阳性。
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引用次数: 18
期刊
The Open Virology Journal
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