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2011年26卷3期

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Review

病毒复制适应性在HIV母婴传播中的作用

陈学庆, 刘畅, 孔晓红

2011, 26(3): 147 doi: 10.1007/s12250-011-3180-2

[HTML全文] [PDF 355 KB] Springerlink

HIV母婴传播指HIV感染母亲将病毒传给婴儿的过程,发生于怀孕,生产和母乳喂养的过程中。全球大约90%的儿童感染源于其母亲,在不采取任何预防措施的情况下,HIV母婴传播的发生率高达30~45%。遗传进化分析和异源双链示踪检测表明,婴儿体内的病毒同源性高于母亲,这是HIV病毒选择性传播的一个重要表现。病毒的选择性传播受母亲和婴儿体内多种因素的影响,HIV病毒复制适应性是近年来备受关注的影响因素之一。它代表了病毒适应宿主环境,进而生存和产生子代病毒的能力。研究表明HIV包膜蛋白gp120是决定传播病毒复制适应性的一个关键因素,说明HIV 自身的遗传背景在HIV 传播过程中有着重要的作用。本文概述了HIV病毒复制适应性在母婴传播中的作用及其研究进展,为HIV 母婴传播的预防,治疗,以及疫苗的研发提供参考。

Characterization of the Receptor-binding Domain of Ebola Glycoprotein in Viral Entry

Jizhen Wang, Balaji Manicassamy, Michael Caffrey, Lijun Rong

2011, 26(3): 156 doi: 10.1007/s12250-011-3194-9

[HTML全文] [PDF 886 KB] Springerlink

Ebola virus infection causes severe hemorrhagic fever in human and non-human primates with high mortality. Viral entry/infection is initiated by binding of glycoprotein GP protein on Ebola virion to host cells, followed by fusion of virus-cell membrane also mediated by GP. Using an human immunodeficiency virus (HIV)-based pseudotyping system, the roles of 41 Ebola GP1 residues in the receptor-binding domain in viral entry were studied by alanine scanning substitutions. We identified that four residues appear to be involved in protein folding/structure and four residues are important for viral entry. An improved entry interference assay was developed and used to study the role of these residues that are important for viral entry. It was found that R64 and K95 are involved in receptor binding. In contrast, some residues such as I170 are important for viral entry, but do not play a major role in receptor binding as indicated by entry interference assay and/or protein binding data, suggesting that these residues are involved in post-binding steps of viral entry. Furthermore, our results also suggested that Ebola and Marburg viruses share a common cellular molecule for entry.

宿主细胞蛋白C9orf69通过与HSV-1 UL25蛋白的作用支持病毒增殖

Ying Zhang, Yan-mei Li, Long-ding Liu, Li Jiang, Ma Ji, Rui-ju Jiang, Lei Guo, Yun Liao and Qi-han Li,

2011, 26(3): 171 doi: 10.1007/s12250-011-3179-8

[HTML全文] [PDF 658 KB] Springerlink

鉴于HSV-1核衣壳蛋白UL25与宿主细胞蛋白相互作用的报道较少,本研究通过酵母双杂交技术寻找能与病毒蛋白UL25相互作用的细胞蛋白,筛选得到功能未知的细胞蛋白C9orf69,并通过荧光共定位、免疫共沉淀等生物实验证实两者在生理状态下存在相互作用。对C9orf69功能的初步研究表明,C9orf69能够促进病毒增殖。进一步研究发现,C9orf69并不是通过调控病毒基因转录水平直接影响病毒的增殖效率,而是该蛋白在与UL25相互作用过程中,间接地促进了病毒增殖速度。

Total Chemical Synthesis, Assembly of Human Torque Teno Virus Genome

Zheng Hou, Gengfu Xiao

2011, 26(3): 181 doi: 10.1007/s12250-011-3187-8

[HTML全文] [PDF 485 KB] Springerlink

Torque teno virus (TTV) is a nonenveloped virus containing a single-stranded, circular DNA genome of approximately 3.8kb. We completely synthesized the 3 808 nucleotides of the TTV (SANBAN isolate) genome, which contains a hairpin structure and a GC-rich region. More than 100 overlapping oligonucleotides were chemically synthesized and assembled by polymerase chain assembly reaction (PCA), and the synthesis was completed with splicing by overlap extension (SOEing). This study establishes the methodological basis of the chemical synthesis of a viral genome for use as a live attenuated vaccine or gene therapy vector.

一种新型Asia 1口蹄疫病毒多表位疫苗对豚鼠的免疫效力研究

邵军军, 王景锋, 常惠芸, 柳纪省

2011, 26(3): 190 doi: 10.1007/s12250-011-3174-0

[HTML全文] [PDF 459 KB] Springerlink

为了研制一种安全高效适用于羊的Asia1型口蹄疫病毒重组亚单位疫苗,选用了口蹄疫病毒的VP1蛋白区的主要抗原表位(137-160和197-211)设计和人工合成了一个串联重复多表位基因,同时引入了具有重要生物学功能的生物大分子,羊免疫球蛋白重链恒定区作为蛋白载体,成功构建了两个原核pET-30a-RE和pET-30a-RE-oIgG,并获得了以包涵体形式表达的重组蛋白。以豚鼠为模型对该疫苗进行了免疫效力评价,结果显示,重要生物功能分子IgG能明显增强表位的免疫效力。重组蛋白RE-oIgG不仅能够诱导高水平的中和抗体和淋巴细胞发生增值反应,而且提供了完全保护。虽然重组蛋白RE不能提供保护,但能延迟疾病出现临川针状的时间,减轻疾病的严重性。因此,我们认为该疫苗研究策略将对未来设计口蹄疫病毒新型疫苗提供新的思路。

建立及其生长特性和对病毒敏感性的研究

吴艳蕾, 姜磊, Yoshifumi Hashimoto, Robert R. Granados, 李国勋

2011, 26(3): 198 doi: 10.1007/s12250-011-3177-x

[HTML全文] [PDF 1284 KB] Springerlink

目前已从鳞翅目sf-9,sf-21和一些家蚕细胞获得多株耐热细胞系,他们在染色体核型,膜脂质组成,形态和生长特性与亲代细胞系有很大的区别。本文报道了其它昆虫种类耐热细胞系的开发,测定 了它们的生长特性和对病毒敏感性。利用通过逐渐分阶段的提高温度,经2个月的处理,成功的获得了耐热细胞系,分别命名为sf9-ht33, sf9-ht35, High5-ht33, High5-ht35, MG1-ht33, MG1-ht35。现已传代70次以上,获得稳定的细胞系。结果表明,耐热细胞系的群体倍增时间比亲代细胞的短1-4小时。细胞形状没有明显变化,而sf9-ht细胞变大,High5和MG1 ht细胞变小。当细胞系在28℃,33℃,35℃,37℃下感染苜蓿银纹夜蛾核型多角体病毒时,芽殖病毒和多角体的产量在相应的细胞系中有最适宜的温度。

新型HPV 16 E6E7 融合基因突变体的转化活性研究

谢强, 张立娜, 徐银胜, 王辛欣

2011, 26(3): 206 doi: 10.1007/s12250-011-3178-9

[HTML全文] [PDF 659 KB] Springerlink

按照哺乳动物细胞密码子偏好性原则优化HPV16的E6和E7基因,并将优化的E6和E7基因进行融合,得到优化融合基因HPV16 ofE6E7。然后对E6的L57G、C113R和E7的C24G、E26G进行定点突变,得到HPV16 E6E7的优化融合基因突变体HPV16 omfE6E7(专利申请号:CN 101100672)。实验研究表明构建的HPV16 omfE6E7基因不仅去除了HPV16 E6 和E7基因的转化活性,而且具有很好的稳定性和较强的抗原活性。这提示,在开发安全有效的HPV16相关肿瘤的治疗性疫苗过程中,HPV16 omfE6E7有望成为理想抗原基因。

猪圆环病毒II型LAMP检测方法的建立

刘业兵, 张磊, 薛青红, 宁宜宝, 张志刚

2011, 26(3): 214 doi: 10.1007/s12250-011-3169-x

[HTML全文] [PDF 457 KB] Springerlink

In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10–5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.