埃博拉出血热及埃博拉病毒的研究进展

埃博拉病毒可以引起一种人畜共患烈性传染病,即埃博拉出血热,此病于1976年始发于埃博拉河流域,并且于该区域严重流行,故而得名。人类一旦感染埃博拉病毒,死亡率可高达88%,从而引起医学界的广泛关注,世界卫生组织已将埃博拉病毒列为对人类危害最为严重的病毒之一。深入地了解埃博拉出血热及埃博拉病毒,及其致病机理,对于埃博拉出血热的预防和控制具有非常重要的意义。     

埃博拉病毒受体的研究进展

埃博拉病毒能在人类和非人灵长类中引起严重的出血热疾病。1976年,国际上首次报道了埃博拉出血热病例,到2014年,非洲又爆发了历史上最严重的埃博拉疫情,然而,至今仍然没有明确的预防疫苗和治疗药物。与大多数病毒一样,埃博拉病毒需要入侵宿主细胞才能完成自身的复制。因此,找到埃博拉病毒入侵的受体并探究其入侵的过程和机制,对于开发病毒疫苗及新型治疗药物具有非常关键的作用。作者希望通过总结埃博拉病毒受体的研究进展,为埃博拉病毒疫苗及治疗药物的开发提供思路。     

埃博拉病毒及其致病机制

自2014年2~3月,西非埃博拉病毒感染的暴发流行已呈播散趋势,受到世界卫生组织的高度重视。我国也提高了防止埃博拉病毒进入国内的警示,并采取了相应措施。现将有关埃博拉病毒的生物学特性、致病机制及相关流行病学与防治策略作简要综述,供参考。     

抗埃博拉病毒药物研究近期动态

对抗埃博拉病毒新药的近期研究成果以及业界提出或可用于治疗埃博拉出血热的已上市药物进行总结归纳,旨在为抗埃博拉病毒药物的进一步开发及埃博拉出血热治疗方案的探索提供参考。     

基于GoPubMed对埃博拉病毒研究文献的数据分析

本研究对埃博拉病毒相关研究载文量的文献计量学分析,以期为埃博拉病毒研究提供文献的数据支持。埃博拉病毒(Ebolavirus,EBOV)是撒哈拉以南的非洲地区高致病性人畜共患传染病,本文基于文献计量学的分析方法,以“Ebolavirus”为主题词检索文献,检索截止至2018年3月2日被PubMed数据库收录的埃博拉病毒(Ebolavirus)相关研究文献,在PubMed中检索到4 391篇文献为研究对象。统计学分析其埃博拉病毒相关研究文献的高频主题词及发表文献的年份、国家、城市和期刊的分布情况,了解埃博拉病毒研究的现状及其发展趋势。结果表明2013-2015年,PubMed数据库收录的埃博拉病毒相关研究的载文量呈逐年上升趋势,2015年至本文统计日的载文量呈现逐年下降。本研究在PubMed数据库检索到的4 391篇埃博拉病毒研究文献中,载文量最多的国家为美国,中国排第六,载文量较多的地区和城市则集中在北美和欧洲等西方发达国家,载文量较多的城市中,中国的北京排第八位。埃博拉病毒相关研究载文量较多的期刊是Journal of Virology(J Virol)。利用GoPubMed检索到的埃博拉病毒相关研究文献,可以从文献数据的角度显示出埃博拉病毒研究的现状和发展趋势。     

埃博拉病毒侵染细胞机制的研究进展

埃博拉病毒能在人类和非人灵长类中引起严重的埃博拉出血热,病死率可高达90%,并且目前还没有针对埃博拉病毒有效的疫苗或者是治疗手段。面对埃博拉病毒带来的挑战,针对埃博拉病毒的相关研究已经成为病毒学的热点问题。其中,关于埃博拉病毒侵染细胞机制的科学研究对于开发病毒疫苗以及新型治疗药物有非常关键的作用。因此,本综述拟就埃博拉病毒侵染细胞机制方面的研究进展进行总结。     

抗流感病毒中药及抗埃博拉病毒活性化合物的发现策略

2014年在非洲爆发流行的埃博拉病毒严重威胁人类健康和生命安全,迄今尚未有治疗药物,目前国内外许多医药机构尝试从不同视角发现抗埃博拉病毒的有效药物.本文在对抗流感病毒中药及活性成分进行回顾的基础上提出了抗埃博拉病毒活性化合物的发现策略,为从中药中发现抗埃博拉病毒有效物质提供借鉴和参考.     

关于埃博拉病毒核酸检测试剂应急审评的探讨

2014年埃博拉病毒出血热疫情引起了全世界的关注,为做好疫情监控,严防疫情输入,药监总局启动埃博拉病毒检测试剂应急审批程序,推动应急用体外诊断试剂产品尽快完成审批。本文就埃博拉病毒核酸检测试剂应急审评要求及审批条件进行解析和介绍,并结合此次应急审批工作提出了对应急审评的思考。     

表达埃博拉病毒Makona突变株糖蛋白的腺病毒载体疫苗在豚鼠和非人灵长类动物体内有效

<正>2014~2016年在西非爆发的埃博拉疫情已导致超过11000人死亡,目前还没有批准使用的埃博拉疫苗。以往研究结果显示,表达埃博拉病毒糖蛋白的重组病毒载体疫苗可以保护非人灵长类动物抵抗致死埃博拉病毒攻毒。然而,这些疫苗表达的糖蛋白来源于在中非流行的埃博拉病毒突变株,与在西非爆发的Makona突变株在氨基酸序列上的同源性为97.3%。本研究中,我们证明了免疫Ad5-Mak GP的豚鼠可产生强烈的体液免疫反应;疫苗完全保护了埃博拉     

人埃博拉病毒密码子偏爱性与聚类分析

埃博拉病毒(Ebola virus,EBOV)宿主广泛,其遗传进化关系复杂,影响埃博拉病毒密码子偏爱性的因素众多。为了明确人埃博拉病毒密码子使用的偏爱性,揭示影响其偏爱性的影响因素和不同宿主来源的埃博拉病毒间的遗传进化关系。本研究通过计算有效密码子数(Effective number of codons,ENC),同义密码子相对使用频率(Relative synonymous codon usage,RSCU)和其他指标,对人埃博拉病毒的密码子使用模式进行综合分析。结果显示,人埃博拉病毒各蛋白的ENC均值分布于55.66～55.77,RSCU1的密码子中77%以上都是以A/U结尾。中性绘图分析和PR2绘图分析等相关分析表明,自然选择是影响密码子使用模式的主要因素,突变压力的影响相对较小。聚类分析结果表明,猪和人来源的埃博拉病毒亲缘关系最近,提示猪来源的埃博拉病毒感染人的风险最大。新发现的bombali型埃博拉病毒与人的亲缘关系距离较远,在大多数蛋白中均单独聚类。本研究结果对深入了解人埃博拉病毒的遗传进化关系,进而研究埃博拉疫苗和制备抗体具有重大意义。     

Protection from Ebola virus mediated by cytotoxic T lymphocytes specific for the viral nucleoprotein

Cytotoxic T lymphocytes (CTLs) are proposed to be critical for protection from intracellular pathogens such as Ebola virus. However, there have been no demonstrations that protection against Ebola virus is mediated by Ebola virus-specific CTLs. Here, we report that C57BL/6 mice vaccinated with Venezuelan equine encephalitis virus replicons encoding the Ebola virus nucleoprotein (NP) survived lethal challenge with Ebola virus. Vaccination induced both antibodies to the NP and a major histocompatibility complex class I-restricted CTL response to an 11-amino-acid sequence in the amino-terminal portion of the Ebola virus NP. Passive transfer of polyclonal NP-specific antiserum did not protect recipient mice. In contrast, adoptive transfer of CTLs specific for the Ebola virus NP protected unvaccinated mice from lethal Ebola virus challenge. The protective CTLs were CD8(+), restricted to the D(b) class I molecule, and recognized an epitope within amino acids 43 to 53 (VYQVNNLEEIC) in the Ebola virus NP. The demonstration that CTLs can prevent lethal Ebola virus infection affects vaccine development in that protective cellular immune responses may be required for optimal protection from Ebola virus.     

Recent progress on the treatment of Ebola virus disease with Favipiravir and other related strategies

Ebola virus is one of the most threatening pathogens with the mortality rate as high as 90% in the world. There are no licensed therapeutic drugs or preventive vaccines for Ebola hemorrhagic fever up to date. Favipiravir, a novel antiviral drug which was mainly used for the treatment of influenza, now has been demonstrated to have a curative effect in treating Ebola virus infection. In this review, we present an overview of recent progress on the treatment of Ebola virus disease with Favipiravir and describe its possible mechanism. Moreover, we give a brief summary of other related treatment strategies against Ebola.     

Successful topical respiratory tract immunization of primates against Ebola virus

Ebola virus causes outbreaks of severe viral hemorrhagic fever with high mortality in humans. The virus is highly contagious and can be transmitted by contact and by the aerosol route. These features make Ebola virus a potential weapon for bioterrorism and biological warfare. Therefore, a vaccine that induces both systemic and local immune responses in the respiratory tract would be highly beneficial. We evaluated a common pediatric respiratory pathogen, human parainfluenza virus type 3 (HPIV3), as a vaccine vector against Ebola virus. HPIV3 recombinants expressing the Ebola virus (Zaire species) surface glycoprotein (GP) alone or in combination with the nucleocapsid protein NP or with the cytokine adjuvant granulocyte-macrophage colony-stimulating factor were administered by the respiratory route to rhesus monkeys--in which HPIV3 infection is mild and asymptomatic--and were evaluated for immunogenicity and protective efficacy against a highly lethal intraperitoneal challenge with Ebola virus. A single immunization with any construct expressing GP was moderately immunogenic against Ebola virus and protected 88% of the animals against severe hemorrhagic fever and death caused by Ebola virus. Two doses were highly immunogenic, and all of the animals survived challenge and were free of signs of disease and of detectable Ebola virus challenge virus. These data illustrate the feasibility of immunization via the respiratory tract against the hemorrhagic fever caused by Ebola virus. To our knowledge, this is the first study in which topical immunization through respiratory tract achieved prevention of a viral hemorrhagic fever infection in a primate model.     

C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans

Ebola virus is a highly lethal pathogen responsible for several outbreaks of hemorrhagic fever. Here we show that the primate lentiviral binding C-type lectins DC-SIGN and L-SIGN act as cofactors for cellular entry by Ebola virus. Furthermore, DC-SIGN on the surface of dendritic cells is able to function as a trans receptor, binding Ebola virus-pseudotyped lentiviral particles and transmitting infection to susceptible cells. Our data underscore a role for DC-SIGN and L-SIGN in the infective process and pathogenicity of Ebola virus infection.     

Protective cytotoxic T-cell responses induced by venezuelan equine encephalitis virus replicons expressing Ebola virus proteins

Infection with Ebola virus causes a severe disease accompanied by high mortality rates, and there are no licensed vaccines or therapies available for human use. Filovirus vaccine research efforts still need to determine the roles of humoral and cell-mediated immune responses in protection from Ebola virus infection. Previous studies indicated that exposure to Ebola virus proteins expressed from packaged Venezuelan equine encephalitis virus replicons elicited protective immunity in mice and that antibody-mediated protection could only be demonstrated after vaccination against the glycoprotein. In this study, the murine CD8(+) T-cell responses to six Ebola virus proteins were examined. CD8(+) T cells specific for Ebola virus glycoprotein, nucleoprotein, and viral proteins (VP24, VP30, VP35, and VP40) were identified by intracellular cytokine assays using splenocytes from vaccinated mice. The cells were expanded by restimulation with peptides and demonstrated cytolytic activity. Adoptive transfer of the CD8(+) cytotoxic T cells protected filovirus na?ve mice from challenge with Ebola virus. These data support a role for CD8(+) cytotoxic T cells as part of a protective mechanism induced by vaccination against six Ebola virus proteins and provide additional evidence that cytotoxic T-cell responses can contribute to protection from filovirus infections.     

Phosphoinositide-3 kinase-Akt pathway controls cellular entry of Ebola virus

The phosphoinositide-3 kinase (PI3K) pathway regulates diverse cellular activities related to cell growth, migration, survival, and vesicular trafficking. It is known that Ebola virus requires endocytosis to establish an infection. However, the cellular signals that mediate this uptake were unknown for Ebola virus as well as many other viruses. Here, the involvement of PI3K in Ebola virus entry was studied. A novel and critical role of the PI3K signaling pathway was demonstrated in cell entry of Zaire Ebola virus (ZEBOV). Inhibitors of PI3K and Akt significantly reduced infection by ZEBOV at an early step during the replication cycle. Furthermore, phosphorylation of Akt-1 was induced shortly after exposure of cells to radiation-inactivated ZEBOV, indicating that the virus actively induces the PI3K pathway and that replication was not required for this induction. Subsequent use of pseudotyped Ebola virus and/or Ebola virus-like particles, in a novel virus entry assay, provided evidence that activity of PI3K/Akt is required at the virus entry step. Class 1A PI3Ks appear to play a predominant role in regulating ZEBOV entry, and Rac1 is a key downstream effector in this regulatory cascade. Confocal imaging of fluorescently labeled ZEBOV indicated that inhibition of PI3K, Akt, or Rac1 disrupted normal uptake of virus particles into cells and resulted in aberrant accumulation of virus into a cytosolic compartment that was non-permissive for membrane fusion. We conclude that PI3K-mediated signaling plays an important role in regulating vesicular trafficking of ZEBOV necessary for cell entry. Disruption of this signaling leads to inappropriate trafficking within the cell and a block in steps leading to membrane fusion. These findings extend our current understanding of Ebola virus entry mechanism and may help in devising useful new strategies for treatment of Ebola virus infection.     

Reverse Genetics Demonstrates that Proteolytic Processing of the Ebola Virus Glycoprotein Is Not Essential for Replication in Cell Culture

Ebola virus, a prime example of an emerging pathogen, causes fatal hemorrhagic fever in humans and in nonhuman primates. Identification of major determinants of Ebola virus pathogenicity has been hampered by the lack of effective strategies for experimental mutagenesis. Here we exploit a reverse genetics system that allows the generation of Ebola virus from cloned cDNA to engineer a mutant Ebola virus with an altered furin recognition motif in the glycoprotein (GP). When expressed in cells, the GP of the wild type, but not of the mutant, virus was cleaved into GP1 and GP2. Although posttranslational furin-mediated cleavage of GP was thought to be an essential step in Ebola virus infection, generation of a viable mutant Ebola virus lacking a furin recognition motif in the GP cleavage site demonstrates that GP cleavage is not essential for replication of Ebola virus in cell culture.     

External Quality Assessment of Molecular Detection of Ebola Virus in China

In 2014, Ebola hemorrhagic fever broke out in West Africa. As contact between China and West Africa is frequent, the possibility that Ebola virus would enter China was high. Thus, an external assessment of the quality of Ebola virus detection was organized by the National Center for Clinical Laboratories in China. Virus-like particles encapsulating known sequences of epidemic strains of Ebola virus from 2014 were prepared as positive quality controls. The sample panel, which was composed of seven positive and three negative samples, was dispatched to 19 laboratories participating in this assessment of Ebola virus detection. Accurate detection was reported at 14 of the 19 participating laboratories, with a sensitivity of 91.43% and a specificity of 100%. Four participants (21.05%) reported false-negative results and were classified as “acceptable.” One participant (5.26%) did not detect any positive samples and was thus classified as “improvable.” Based on the results returned, the ability to detect weakly positive Ebola specimens should be improved. Furthermore, commercial assays and the standard primers offered by the Chinese Centers for Disease Control and Prevention were found to be most accurate and dependable for Ebola detection. A two-target detection approach is recommended for Ebola screening; this approach could reduce the probability of false-negative results. Additionally, standardization of operations and punctual adjustment of instruments are necessary for the control and prevention of Ebola virus.