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2016年31卷6期

Viral pathogens frequently hijack the host actin polymerization machinery to facilitate their infection. Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) is one of the unique insect virus that inducing actin polymerization occurring in the nucleus, which requires relocation of cytoplasmic actin polymerization machinery, including actin-related protein 2/3 complex (Arp2/3), to the nucleus. Therefore, the alphabaculovirus-infection system could serve as a research model for investigating the nuclear import mechanism of Arp2/3. In this issue, Wang et al. investigated the spatial changes of Arp2/3 subunits induced by the viral replication-associated protein Ac34 in Sf9 cells. The cover shows the relocation of Arp2/3 subunits (P40, Arp2, P34, P21, and P20) in the presence of the viral late gene product Ac34. See page 472-479 for details.

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Review

激酶IKKε在天然免疫和获得性免疫中的作用概述

张军杰, 田茂, 夏赞贤, 冯平辉

2016, 31(6): 457 doi: 10.1007/s12250-016-3898-y

[HTML全文] [PDF 1297 KB] Springerlink
IKKε作为一种非经典的IκB激酶,在天然免疫中被广泛研究。最近的研究发现IKKε在干扰素信号通路中发挥重要作用。同时, 最新的研究发现IKKε在获得性免疫中也具有重要的功能。IKKε负调控CD8 T 细胞激活,暗示IKKε可能是一个潜在的免疫治疗的靶点,可以通过抑制IKKε来提高抗病毒和抗肿瘤T细胞免疫。我们概述了IKKε在干扰素信号通路及T细胞免疫中的研究进展,同时提出了亟待解决的相关学术问题。

卡波氏肉瘤相关疱疹病毒对机体双链DNA识别系统的免疫逃逸机制

高航, 宋艳艳, 刘成容, 梁启明

2016, 31(6): 466 doi: 10.1007/s12250-016-3877-3

[HTML全文] [PDF 780 KB] Springerlink
天然免疫系统通过模式识别受体cGAS识别细胞内双链DNA来启动1型干扰素通路和自噬通路,二者协同作用抑制病原体感染并激活机体的细胞免疫应答。疱疹病毒的基因组属于双链DNA,在感染中别cGAS识别。然而,为了成功的进行持续性感染,疱疹病毒已进化出多个病毒基因来调节宿主免疫信号通路的不同方面。本综述详细总结了卡波氏肉瘤相关疱疹病毒对机体双链DNA识别系统的免疫逃逸机制以及其对KSHV生命周期的意思。

微小RNA分子在肝细胞代谢和乙肝病毒复制中的调控作用

邓万玉, 陆蒙吉

2016, 31(6): 472 doi: 10.1007/s12250-016-3924-0

[HTML全文] [PDF 469 KB] Springerlink
目前,虽然已有较高效的预防性疫苗和抗病毒治疗方法,但乙型肝炎病毒(hepatitis B virus, HBV)感染,特别是慢乙肝感染仍然是一个全球性的健康问题。虽然对病毒的感染方式和它的复制周期已经有相对比较明确的认识,但是当前对病毒感染诱发的宿主细胞代谢异常的机制以及参与调控病毒复制的宿主因子还有待进一步挖,因此,通过对 HBV-宿主相互作用的研究可以为抗病毒治疗提供一定的理论支持。微小RNA(microRNA,miRNA)是最近几年发展起来的一类转录后调节性小RNA分子,它们参与调控很多胞内活动及信号通路,包括细胞生长代谢和病毒复制。 在这篇综述中,我们以病毒-宿主细胞之间的相互作用为宗旨,旨在探讨微小RNA分子在HBV复制和宿主细胞代谢中的调控作用。
Research Article

The role of viral protein Ac34 in nuclear relocation of subunits of the actin-related protein 2/3 complex

2016, 31(6): 480 doi: 10.1007/s12250-016-3912-4

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The actin nucleator actin-related protein complex (Arp2/3) is composed of seven subunits: Arp2,Arp3, p40/ARPC1 (P40), p34/ARPC2 (P34), p21/ARPC3 (P21), p20/ARPC4 (P20), and p16/ARPC5(P16). Arp2/3 plays crucial roles in a variety of cellular activities through regulation of actin polymerization. Autographa californica multiple nucleopolyhedrovirus (AcMNPV), one of the beststudied alphabaculoviruses, induces Arp2/3 nuclear relocation and mediates nuclear actin polymerization to assist in virus replication. We have demonstrated that Ac34, a viral late-gene product, induces translocation of the P40 subunit of Arp2/3 to the nucleus during AcMNPV infection. However, it remains unknown whether Ac34 could relocate other Arp2/3 subunits to the nucleus. In this study, the effects of the viral protein Ac34 on the distribution of these subunits were studied by an immunofluorescence assay. Arp2, P34, P21, and P20 cloned from Spodoptera frugiperda (Sf9) cells showed mainly cytoplasmic localization and were relocated to the nucleus in the presence of Ac34. In addition, Arp3 was localized in the cytoplasm in both the presence and absence of Ac34, and P16 showed whole-cell localization. In contrast to Sf9 cells, all subunits of mammalian Arp2/3 showed no nuclear relocation in the presence of Ac34. Co-immunoprecipitation analysis of the interaction between Ac34 and Arp2/3 subunits revealed that Ac34 bound to P40, P34, and P20 of Sf9 cells. However, none of the subunits of mammalian Arp2/3 interacted with Ac34, indicating that protein-protein interaction is essential for Ac34 to relocate Arp2/3 subunits to the nucleus.

Characterization of two monoclonal antibodies, 38F10 and 44D11, against the major envelope fusion protein of Helicoverpa armigera nucleopolyhedrovirus

2016, 31(6): 490 doi: 10.1007/s12250-016-3831-4

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The envelope fusion protein F of baculoviruses is a class I viral fusion protein which play a significant role during virus entry into insect cells. F is initially synthesized as a precursor (F0) and then cleaved into a disulfide-linked F1 and F2 subunits during the process of protein maturation and secretion. To facilitate further investigation into the structure and function of F protein during virus infection, monoclonal antibodies (mAbs) against the F2 subunit of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) (HaF) were generated. Two kinds of mAbs were obtained according to their different recognition epitopes: one kind of mAbs, as represented by 38F10, recognizes amino acid (aa) 85 to 123 of F2 and the other kind, represented by 44D11, recognizes aa 148 to 173 of F2. Western blot and immunofluorescence assay confirmed that both of the mAbs recognized the F protein expressed in HearNPV infected cells, however, only 44D11 could neutralize HearNPV infection. The results further showed that 44D11 may not interact with a receptor binding epitope, rather it was demonstrated to inhibit syncytium formation in cells expressing the HaF protein. The results imply that the monoclonal antibody 44D11 recognizes a region within HaF2 that may be involved in the F-mediated membrane fusion process.

埃博拉病毒糖蛋白在大肠杆菌内的成功表达

宰俊杰, 伊尹华, 夏菡, 张波, 袁志明

2016, 31(6): 500 doi: 10.1007/s12250-016-3845-y

[HTML全文] [PDF 14835 KB] Springerlink
埃博拉病毒(EBOV)主要感染人和灵长目动物,可引起严重的出血热,致死率最高可达90%。糖蛋白(GP)是埃博拉病毒唯一的囊膜蛋白,在病毒的吸附和侵入过程中发挥着重要的作用,并且可以刺激机体产生保护性免疫反应。然而至今,埃博拉病毒GP蛋白仍旧无法在大肠杆菌内成功表达,这也阻碍了对病毒蛋白功能的进一步深入研究。本研究中,将埃博拉病毒的基质蛋白40基因(VP40)和GP基因融合在一起,并克隆到原核表达载体中,结果显示,在16℃诱导温度下,VP40-GP和GP-VP40融合蛋白可以在大肠杆菌内表达。此外,结果表明VP40基因在融合蛋白中的位置,极大地影响了融合蛋白的产量;当VP40基因位于GP基因的上游时,融合蛋白的产量更高。总之,我们的实验结果提供了一种可以大量表达埃博拉病毒全长GP蛋白的方法,为将来研究GP蛋白结构和功能之间的关系,和研制埃博拉治疗性抗体奠定良好的基础。
Letter

细胞热休克蛋白90和卡波齐肉瘤相关疱疹病毒编码的蛋白激酶ORF36相互作用中起重要作用的氨基酸的研究

王超, 王敬超, 郭卫, 龙聪, 汪欢, 孙晓平

2016, 31(6): 509 doi: 10.1007/s12250-016-3839-9

[HTML全文] [PDF 2171 KB] Springerlink ESM
蛋白-蛋白相互作用在生物过程中伴有非常重要的角色。拆分荧光素酶互补试验(SRLCA)是研究蛋白间相互作用的方法之一。该技术是将荧光蛋白在合适的位点切开形成不发荧光的2个片段,这2个片段借助融合于其上的目标蛋白的相互作用,彼此靠近,重新形成能具有活性的荧光蛋白。卡波齐肉瘤相关疱疹病毒(KSHV)编码的蛋白激酶(ORF36)在其相关肿瘤的发生发展过程中起重要作用。同时,细胞内的热休克蛋白90(Hsp90)在各种生物过程中也起到极其重要的作用。该研究中,我们用IP和SRLCA证明Hsp90和ORF36可以在细胞中相互作用。ORF36 215-257氨基酸区段中M216, L221, D225, F226, W237, T238和Hsp90 110-180氨基酸区段中A121, Q133, F134, T149, E158, G168在其相互作用中其关键作用。这些结果可以给我们在治疗KSHV相关疾病中提供一些治疗策略。

Detection and pathogenesis of a novel swine H3N2 influenza virus containing three genes from the 2009 pandemic H1N1 influenza viruses in Korea in 2015

2016, 31(6): 513 doi: 10.1007/s12250-016-3848-8

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On July 2, 2015, four pigs with severe respiratory distress and high fever (>40.0 ℃) at a farm in Jeonpook province in the Southern region of South Korea were euthanized, and their lung tissues were collected and homogenized in PBS. The homogenized samples were inoculated into the wells of 6-well plates containing MadinDarby Canine Kidney (MDCK) cells in MEM with 1 μg/mL trypsin. One influenza A virus, A/Swine/Korea/S2001/2015 (H3N2), was isolated from one sample.

Primary EBV infection and hypersensitivity to mosquito bites: acase report

2016, 31(6): 517 doi: 10.1007/s12250-016-3868-4

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Here, we report what we believe is the first case of EBV-associated HMB from Europe. Follow-up schedules and the necessity of reporting of EBV-associated HMB cases are being discussed.

一种快速拯救基因VII型新城疫病毒的方法的建立

孙玉章, 孙明军, 代永联, 尹仁福, 丁壮

2016, 31(6): 521 doi: 10.1007/s12250-016-3869-3

[HTML全文] [PDF 1602 KB] Springerlink
反向遗传学是研究新城疫病毒(NDV)基因功能、致病机制和新型疫苗候选株的有力工具。基于T7 RNA聚合酶建立的标准新城疫病毒反向遗传操作系统是通过锤头核酶(HamRz)和丁型肝炎病毒核酶(HdvRz)的自催化活性实现病毒反义基因组5'-末端和3'-末端的精确转录。本研究利用具有增强的自剪切活性的优化HdvRz序列,建立起了一种快速拯救基因VII型新城疫病毒的方法。除了提高了拯救新城疫病毒的效率之外,用于检测的微型基因组的报告基因的表达也显著提高。本研究中建立的改进的NDV反向遗传学系统将有助于促进鹅源NDV的传播机理、致病机制、病毒载体和新型疫苗等方向的研究。

中国四种烟草花叶病毒的分子多样性及分布

吴宽, 陈伟, 罗朝鹏, 王冰, 成巨龙, 康振生

2016, 31(6): 525 doi: 10.1007/s12250-016-3728-2

[HTML全文] [PDF 1162 KB] Springerlink ESM
植物病毒病是我国烟草安全生产的重大威胁。为深入研究烟草四种病毒的分布、分子变异,本研究开展了烟草病毒病普查,在2010和2011年采集409份样品。RT-PCR方法用于检测这些样品,结果表明271份样品至少被一种病毒感染:TMV在221个样品中检测到, CMV在132个样品, PVY在86个样品,TEV在62个样品. 从阳性样品中,18 TMV 分离物, 21 CMV分离物, 21 PVY分离物, 15 TEV分离物被鉴定得到并用于多样性分析。基于外壳蛋白全球TMV、CMV、TEV和PVY多样分析表明TMV、CMV、PVY被分离成2群组,TEV分离成4个群组。本文系统研究我国四种烟草病毒分布及遗传多样性,研究结果将为我国制定合理的烟草病毒病防治策略提供重要的理论基础。