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2008 Vol.23(5)

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Current Status of Natural Products from Plants as Anti-herpes Simplex Virus 1 Agents

Yang-fei XIANG, Ying PEI, Yi-fei WANG

2008, 23(5): 305 doi: 10.1007/s12250-008-2962-7

Received: 26 March 2008 Accepted: 03 July 2008
Abstract: Nucleoside analogues have been the mainstay of clinical treatment of herpes simplex virus 1 (HSV-1) infections since their development. However, the emergence of drug resistant strains has underlined the urgency of the discovery of novel anti-HSV-1 drugs. Natural products, which provided many novel drug leads, are known to be an important source of anti-HSV-1 agents. Herein, we present an overview of natural products with anti-HSV-1 activities isolated from a variety of plants reported in recent years. Several different compounds, mainly belonging to the three groups of polysaccharides, polyphenols and terpenes, showed antiviral effects against HSV-1, indicating their potential to be promising anti-HSV-1 agents

Aptamers Against Viral Hepatitis: from Rational Design to Practical Application*

Hui FENG, Kang-hong HU

2008, 23(5): 315 doi: 10.1007/s12250-008-2979-y

Received: 18 June 2008 Accepted: 03 August 2008
Abstract: Aptamers are short nucleic acids or peptides that strongly bind to a protein of interest and functionally inhibit a given target protein at the intracellular level. Besides high affinity and specificity, aptamers have several advantages over traditional antibodies. Hence, they have been broadly selected to develop antiviral agents for therapeutic applications against hepatitis B and C viruses (HBV, HCV). This review provides a summary of in vitro selection and characterization of aptamers against viral hepatitis, which is of practical significance in drug discovery.

The ORF 113 of Heliocoverpa armigera Single Nucleopolyhedrovirus Encodes a Functional Fibroblast Growth Factor*

Xiang LI, Chang-yong LIANG, Jian-hua SONG, Xin-wen CHEN

2008, 23(5): 321 doi: 10.1007/s12250-008-2969-0

Received: 30 April 2008 Accepted: 12 May 2008
Abstract: Fibroblast growth factor (FGF) is a key regulator of developmental processes. A FGF homolog (vFGF) is found in all lepidopteran baculoviruses. Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) vFGFs are chemotactic factors. Here we analyzed the vfgf of Helicoverpa armigera NPV (HearNPV), a group II NPV. The HearNPV vfgf transcripts were detected from 18 to 96 h post-infection (hpi) of Hz-AM1 cells with HearNPV and encoded a 36 kDa protein, which was secreted into the culture medium. HearNPV vFGF had strong affinity to heparin, a property important for FGF signaling via an FGF receptor. Unlike its AcMNPV homolog, HearNPV vFGF specially chemoattracted Hz-AM1, but not other insect cells such as Sf9 and Se-UCR and not the mammalian cells 293 and HepG2. HearNPV vFGF is also associated with the envelope of BV but is absent in occlusion-derived virus, which coordinated to the chemotatic activity analysis.

Preliminary Study on a Potential Panel for Quality Assurance of ELISPOT

Chun-tao ZHANG, Yu WU, Chen-yan ZHAO, Kun-xue HONG, Chun-yu LIU, Ying WANG, Ping ZHONG, Jian-hui NIE, Xue-lin WU, You-chun WANG

2008, 23(5): 330 doi: 10.1007/s12250-008-2933-2

Received: 15 January 2008 Accepted: 29 July 2008
Abstract: The ELISpot assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories

Silencing of UBP43 by shRNA Enhances the Antiviral Activity of Interferon against Hepatitis B Virus*

He-bin FAN, Bao-ju WANG, Yin-ping LU, You-hua HAO, Xin-xing YANG, Meng-ji LU, Dong-liang YANG

2008, 23(5): 339 doi: 10.1007/s12250-008-2960-9

Received: 24 March 2008 Accepted: 20 July 2008
Abstract: Previous studies have shown that expression of the interferon-sensitive gene (ISG)15 protease UBP43 is increased in the liver biopsy specimens of patients who do not respond to interferon (IFN)-α therapy. We hypothesized that UBP43 might hinder the ability of IFN to inhibit HBV replication. In this study, we investigated whether vector-based siRNA promoted by H1(psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. UBP43 was specifically silenced using shRNA. In HepG2.2.15 cells, the HBeAg and HBV DNA levels were significantly reduced by IFN after transfection of shRNA, so vector-based siRNA promoted by H1 (psiUBP43) could enhance IFN inhibiting HBV replication in cell culture. These data suggest that UBP43 modulates the anti-HBV type I IFN response, and is a possible therapeutic target for the treatment of HBV infection.

An Improved Culture System for Virus Isolation and Detection*

Yu-chen XIA, Zhi-hong HU, Zhi-juan QIU, Zhong-bin MA, Hua-lin WANG, Fei DENG

2008, 23(5): 345 doi: 10.1007/s12250-008-2968-1

Received: 14 April 2008 Accepted: 25 July 2008
Abstract: Cell culture has played an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology study of viruses. In the present study, a new system that could support multiple different cell lines to be simultaneous cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called isolated co-cultured system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.

Secretory Expression of E2 Main Antigen Domain of CSFV C Strain and the Establishment of Indirect ELISA Assay*

Guo-zhen LIN, Chang-qing QIU, Fu-ying ZHENG, Ji-zhang ZHOU, Xiao-an CAO

2008, 23(5): 363 doi: 10.1007/s12250-008-2970-7

Received: 05 May 2008 Accepted: 18 August 2008
Abstract: The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus(DE3)–RIL using the pGEX-4T-1 expression vector and the soluble recombinant product purified with Glutathione Sepharose TM4B by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 g/mL and the optimal dilution of serum was 180. The positive cut-off value of this ELISA assay was OD tested serum / OD negative serum≥2.1. The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.

Expression of Endogenous Retrovirus ev/J gp85 Gene and Analysis of Its Immunoreactivity in Comparison with Exogenous Viral Protein*

Yu-ying YANG, Ai-jian QIN, Xiong-yan LIANG, Shu-mei TONG

2008, 23(5): 369 doi: 10.1007/s12250-008-2971-6

Received: 08 May 2008 Accepted: 26 August 2008
Abstract: The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).

Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asia1 by RT-PCR*

Jun-jun SHAO, Hui-yun CHANG, Tong LIN, Guo-zheng CONG, Jun-zheng DU, Jian-hong GUO, Hui-fang BAO, You-jun SHANG, Ya-min YANG, Xiang-tao LIU, Zai-xin LIU, Ji-xing LIU

2008, 23(5): 378 doi: 10.1007/s12250-008-2980-5

Received: 18 June 2008 Accepted: 12 August 2008
Abstract: To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.