For best viewing of the website please use Mozilla Firefox or Google Chrome.




向阳飞, 裴赢, 王一飞

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


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


Hui FENG, hong HU

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


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.


李祥, 梁昌镛, 宋建华, 陈新文

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

FGF是一个非常重要的调节因子,它能影响许多细胞的生长、发育和迁移。在鳞翅目杆状病毒中发现了一个FGF同源家族(vFGF)。已经报道AcMNPV和BmNPV的vFGF具有趋化能力,可能在口服感染中起一定作用。本文主要研究棉铃虫核多角体病毒(HearNPV)vFGF的基本特性及其功能。RT-PCR结果发现HearNPV vfgf 从感染后的18 h开始转录,转录水平随时间延长而持续增加,在感染96 h后还能检测到。对病毒感染不同时间的细胞进行Western blotting检测发现,在感染后24 h到96 h都能检测到一条36 kDa特异带;同时在病毒感染细胞的上清中也检测到了vHaFGF的存在,表明vHaFGF能分泌到细胞外,暗示了它作为胞外的配体能跟肝素结合在信号传导过程中起作用。趋化实验结果显示,HearNPV vFGF只能特异地趋化Hz-AM1细胞,而对其它的Sf9 、Se-UCR、293和HepG2细胞没有趋化能力,这与AcMNPV和BmNPV的vFGF不同,它们能趋化多种不同来源的昆虫细胞。另外,我们还发现vHaFGF作为BV的囊膜蛋白而存在,而ODV上没有,这与BV的趋化实验的结果一致,同时也意味着vHaFGF在BV的感染中起作用

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.


张春涛, 吴瑜, 赵晨燕, 洪坤学, 刘春雨, 王盈, 钟平, 聂建辉, 吴雪伶, 王佑春

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


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.


樊和斌, 王宝菊, 卢银平, 郝友华, 杨新星, 陆蒙吉, 杨东亮

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

以前的研究发现对干扰素无应答的患者肝内干扰素刺激基因(ISG)15的蛋白酶UBP43上升 。因此推测UBP43可能阻碍干扰素抑制HBV复制的作用。本研究在HepG2.2.15细胞中,观察了通过载体介导的shRNA沉默UBP43是否能增加干扰素抑制HBV复制的作用。结果显示HBeAg的表达和HBV DNA的复制显著下降,干扰素的抗病毒活性增强。因此,UBP43影响干扰素的抗病毒活性,是治疗HBV感染的一个可能靶点。

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.


夏宇尘, 丘志娟, 马仲彬, 王华林, 胡志红, 邓菲

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


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.

猪瘟病毒C株E2基因主要抗原区的分泌型表达 及间接ELISA方法的建立

蔺国珍, 邱昌庆*, 郑福英, 周继章, 曹小安

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

将猪瘟病毒C株E2基因主要抗原区原核表达载体pGEX-E2转化进大肠杆菌BL21–CodonPlus(DE3)–RIL中,经条件优化,获得可溶性表达的目的蛋白。应用Glutathione Sepharose TM4B介质纯化重组目的蛋白,Western blot试验表明纯化的目的蛋白能被猪瘟病毒阳性血清识别,具有很好的免疫原性。以纯化的目的蛋白作为包被抗原,初步建立了检测猪瘟病毒抗体的iE2-ELISA诊断方法。结果表明,抗原的最佳包被浓度为0.6 g/mL,血清的最佳稀释度为180。阳性标准为待检血清OD 490 / 阴性血清OD 490≥2.1。应用iE2-ELISA和猪瘟间接血凝试验对100份田间血清样品检测,结果表明两种方法的相对敏感性和特异性分别为90.3% 和 94.7%。特异性实验表明,牛病毒性腹泻血清和猪瘟E2蛋白间没有交叉反应

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.

内源性反转录病毒ev/J gp85基因表达及其免疫活性分析

杨玉莹, 秦爱建, 梁雄燕, 童淑梅

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

内源性反转录病毒ev/J是新近鉴定的内源性禽反转录病毒序列,其gp85基因与ALV-J禽白血病病毒具有高度的同源性。本研究利用英杰Bac-toBac杆状病毒表达系统表达了源自商品肉鸡的ev/J gp85基因。使用ALV-J特异性单克隆抗体JE9 (ADOL-4817)、抗ev/J gp85基因编码物(SU)阳性鼠血清和ALV-J自然感染鸡阳性血清,通过免疫荧光、Western blot、间接ELISA和阻断ELISA分析了重组内源性ev/J gp85 SU的免疫原性和免疫反应性。结果表明ev/J SU可以特异识别JE9单抗和ALV-J自然感染鸡阳性血清,而且抗ev/J SU鼠血清能够阻断外源性ALV-J gp85 SU与ALV-J自然感染鸡阳性血清之间的免疫反应。结果表明内源性ev/J gp85基因重组表达物与外源性ALV-J囊膜蛋白(ADOL-4817 and IMC10200)之间有紧密的免疫学相关性。

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).


邵军军, 常惠芸*, 林 彤, 丛国正, 独军政, 郭建宏, 包慧芳, 尚佑军, 杨亚明, 刘湘涛, 刘在新, 柳纪省

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


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.