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2013年28卷1期

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News & Views

How the Key Finds its Door-Identification of HBV Receptor

Yun Wang, Xinwen Chen

2013, 28(1): 1 doi: 10.1007/s12250-013-3301-1

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HBV is the predominant pathogen associated with hepatitis cases in China. Although the HBV replication mechanism has been extensively documented in recent years, the virus entry mechanism remains elusive; in particular, the HBV receptor has yet to be identified. Recently, a research team led by Dr. Wenhui Li finally identified Sodium taurocholate cotransporting polypeptide (NTCP) as the receptor for HBV infection of hepatocytes (Yan H, et al., 2012). This review highlights their research strategy, as well as the significance of the identification of the HBV receptor.

Review

An Overview of the Highly Pathogenic H5N1 Influenza Virus

Jingchuan Yin, Shi Liu, Ying Zhu

2013, 28(1): 3 doi: 10.1007/s12250-013-3294-9

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Since the first human case of H5N1 avian influenza virus infection was reported in 1997, this highly pathogenic virus has infected hundreds of people around the world and resulted in many deaths. The ability of H5N1 to cross species boundaries, and the presence of polymorphisms that enhance virulence, present challenges to developing clear strategies to prevent the pandemic spread of this highly pathogenic avian influenza (HPAI) virus. This review summarizes the current understanding of, and recent research on, the avian influenza H5N1 virus, including transmission, virulence, pathogenesis, clinical characteristics, treatment and prevention.

Research Article

Mutagenesis of D80-82 and G83 Residues in West Nile Virus NS2B: Effects on NS2B-NS3 Activity and Viral Replication

Fan Jia, Jingjing Fan, Bo Zhang, Zhiming Yuan

2013, 28(1): 16 doi: 10.1007/s12250-013-3276-y

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Flaviviral NS2B is a required cofactor for NS3 serine protease activity and plays an important role in promoting functional NS2B-NS3 protease configuration and maintaining critical interactions with protease catalysis substrates. The residues D80DDG in West Nile virus (WNV) NS2B are important for protease activity. To investigate the effects of D80DDG in NS2B on protease activity and viral replication, the negatively charged region D80DD and the conserved residue G83 of NS2B were mutated (D80DD/E80EE, D80DD/K80KK, D80DD/A80AA, G83F, G83S, G83D, G83K, and G83A), and NS3 D75A was designated as the negative control. The effects of the mutations on NS2B-NS3 activity, viral translation, and viral RNA replication were analyzed using kinetic analysis of site-directed enzymes and a transient replicon assay. All substitutions resulted in significantly decreased enzyme activity and blocked RNA replication. The negative charge of D80DD is not important for maintaining NS2B function, but side chain changes in G83 have dramatic effects on protease activity and RNA replication. These results demonstrate that NS2B is important for viral replication and that D80DD and G83 substitutions prevent replication; they will be useful for understanding the relationship between NS2B and NS3.

Detection of the Pandemic H1N1/2009 Influenza A Virus by a Highly Sensitive Quantitative Real-time Reverse-transcription Polymerase Chain Reaction Assay

Zhu Yang, Guoliang Mao, Yujun Liu, Yuan-Chuan Chen, Chengjing Liu, Jun Luo, Xihan Li, Ke Zen, Yanjun Pang, Jianguo Wu, Fenyong Liu

2013, 28(1): 24 doi: 10.1007/s12250-013-3290-0

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A quantitative real time reverse-transcription polymerase chain reaction (qRT-PCR) assay with specific primers recommended by the World Health Organization (WHO) has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin (HA) genes of the pandemic H1N1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic H1N1/2009 virus infection.

An Inactivated Vaccine from a Field Strain of Bovine Herpesvirus-1 (BoHV-1) has High Antigenic Mass and Induces Strong Efficacy in a Rabbit Model

Julian Ruiz-Sáenz, Jairo Jaime, Victor Vera

2013, 28(1): 36 doi: 10.1007/s12250-013-3283-z

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Bovine Herpesvirus-1 (BoHV-1) is a DNA virus belonging to the family Herpesviridae, subfamily Alfaherpesvirinae; it is a worldwide pathogen, causing serious economic losses in livestock. In Colombia there have been multiple isolates of BoHV-1 that have been subjected to molecular characterization, classifying most of the country isolates as BoHV-1.1. In the present study we developed and evaluated an ethyleneimine binary inactivated isolate from the native BoHV-1 strain (Córdoba-2) in a rabbit model of vaccination and infection. The vaccine was evaluated in two phases, one of immunogenicity with vaccination and a booster after 21 days, and an evaluation phase of protection against challenge with a highly virulent reference strain. The results demonstrate optimum serum-conversion, with protective neutralizing antibody titers 28 days post vaccination and optimal protection against challenge with the reference strain with decreased clinical signs of infection, protection against the onset of fever and decrease of virus excretion post challenge. In conclusion, our results show the enormous potential that an immunogenic inactivated vaccine has produced from the native BoHV-1.1 strain, which produces a high antigen mass to the vaccine to induce optimal immunity and protection, and it is a strong candidate for evaluation and possible future use in different cattle populations.

Modification in Media Composition to Obtain Secretory Production of STxB-based Vaccines using Escherichia coli

Mohammad Sadraeian, Mohammad Bagher Ghoshoon, Milad Mohkam, Zeinab Karimi, Sara Rasoul-Amini, Younes Ghasemi

2013, 28(1): 43 doi: 10.1007/s12250-013-3286-9

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Shiga toxin B-subunit (STxB) from Shigella dysenteriae targets in vivo antigen to cancer cells, dendritic cells (DC) and B cells, which preferentially express the globotriaosylceramide (Gb3) receptor. This pivotal role has encouraged scientists to investigate fusing STxB with other clinical antigens. Due to the challenges of obtaining a functional soluble form of the recombinant STxB, such as formation of inclusion bodies during protein expression, scientists tend to combine STxB with vaccine candidates rather than using their genetically fused forms. In this work, we fused HPV16 E7 as a vaccine candidate to the recombinantly-produced STxB. To minimize the formation of inclusion bodies, we investigated a number of conditions during the expression procedure. Then various strategies were used in order to obtain high yield of soluble recombinant protein from E. coli which included the use of different host strains, reduction of cultivation temperature, as well as using different concentrations of IPTG and different additives (Glycin, Triton X-100, ZnCl2). Our study demonstrated the importance of optimizing incubation parameters for recombinant protein expression in E. coli; also showed that the secretion production can be achieved over the course of a few hours when using additives such as glycine and Triton X-100. Interestingly, it was shown that when the culture mediums were supplemented by additives, there was an inverse ratio between time of induction (TOI) and the level of secreted protein at lower temperatures. This study determines the optimal conditions for high yield soluble E7-STxB expression and subsequently facilitates reaching a functionally soluble form of STxB-based vaccines, which can be considered as a potent vaccine candidate for cervical cancer.

Letter

A Case of Hepatitis B Reactivation in an Anti-HBs Positive, Anti-HBc Positive non-Hodgkin’s Lymphoma Patient

Chunchen Wu, Hui Shi, Mengji Lu, Yang Xu, Xinwen Chen

2013, 28(1): 49 doi: 10.1007/s12250-013-3285-x

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Molecular Characterizations of Two Begomoviruses Infecting Vinca rosea and Raphanus sativus in India

Avinash Marwal, Anurag Sahu, Pradeep Sharma, Rajarshi Kumar Gaur

2013, 28(1): 53 doi: 10.1007/s12250-013-3275-z

[HTML全文] [PDF 458 KB] Springerlink