埃博拉病毒入侵宿主细胞的分子机制

 埃博拉病毒是一类能够感染并引起人和灵长类动物发生埃博拉出血热的烈性囊膜病毒。发现近40年中,埃博拉病毒给人类生命带来了极大威胁。然而,目前人们对于埃博拉病毒的了解非常有限,尤其是病毒与其宿主细胞受体的结合机制和膜融合机制相关信息的缺失,使得针对埃博拉病毒的特效药物的设计和研发工作阻碍重重。本文综述了埃博拉病毒分类、形态、病毒蛋白和病毒生命周期,着重介绍了高福院士团队在埃博拉病毒入侵宿主细胞的分子机制研究中的成果。通过结构学手段解析了埃博拉病毒激活态囊膜糖蛋白GPcl与宿主细胞受体NPC1分子的复合物结构,从原子水平上阐明了埃博拉病毒与宿主细胞相互识别的机制,并在结构基础上对病毒的膜融合促发机制做出推测,提出以埃博拉病毒为代表的新的（第5种）囊膜病毒膜融合激发机制,为抗埃博拉病毒药物和疫苗的设计提供了结构基础。     

Development of a preventive vaccine for Ebola virus infection in primates

Outbreaks of haemorrhagic fever caused by the Ebola virus are associated with high mortality rates that are a distinguishing feature of this human pathogen. The highest lethality is associated with the Zaire subtype, one of four strains identified to date. Its rapid progression allows little opportunity to develop natural immunity, and there is currently no effective anti-viral therapy. Therefore, vaccination offers a promising intervention to prevent infection and limit spread. Here we describe a highly effective vaccine strategy for Ebola virus infection in non-human primates. A combination of DNA immunization and boosting with adenoviral vectors that encode viral proteins generated cellular and humoral immunity in cynomolgus macaques. Challenge with a lethal dose of the highly pathogenic, wild-type, 1976 Mayinga strain of Ebola Zaire virus resulted in uniform infection in controls, who progressed to a moribund state and death in less than one week. In contrast, all vaccinated animals were asymptomatic for more than six months, with no detectable virus after the initial challenge. These findings demonstrate that it is possible to develop a preventive vaccine against Ebola virus infection in primates.     

Accelerated vaccination for Ebola virus haemorrhagic fever in non-human primates

Containment of highly lethal Ebola virus outbreaks poses a serious public health challenge. Although an experimental vaccine has successfully protected non-human primates against disease, more than six months was required to complete the immunizations, making it impractical to limit an acute epidemic. Here, we report the development of accelerated vaccination against Ebola virus in non-human primates. The antibody response to immunization with an adenoviral (ADV) vector encoding the Ebola glycoprotein (GP) was induced more rapidly than with DNA priming and ADV boosting, but it was of lower magnitude. To determine whether this earlier immune response could nonetheless protect against disease, cynomolgus macaques were challenged with Ebola virus after vaccination with ADV-GP and nucleoprotein (NP) vectors. Protection was highly effective and correlated with the generation of Ebola-specific CD8(+) T-cell and antibody responses. Even when animals were immunized once with ADV-GP/NP and challenged 28 days later, they remained resistant to challenge with either low or high doses of virus. This accelerated vaccine provides an intervention that may help to limit the epidemic spread of Ebola, and is applicable to other viruses.     

埃博拉病毒感染数量的一个数学模型

埃博拉病毒病(EVD)是严重的、往往致命的人类疾病,病死率高达90%.埃博拉病毒病疫情主要发生在中非和西非靠近热带雨林的边远村庄.该病毒通过野生动物传到人,并且通过人际间传播在人群中蔓延.病情严重的患者需要获得重症支持治疗,无论对人还是对动物都无可用的已获正式许可的特异性治疗办法或者疫苗.由于缺乏有效的治疗手段和人用疫苗,提高对感染埃博拉危险因素的认识以及个人可以采取一些保护措施,这是减少人类感染和死亡的唯一方法.本文建立一个埃博拉病毒的数学模型,对疫情进行实证分析;并且对疫情的发展也做了一个预测.     

Ebola virus entry requires the cholesterol transporter Niemann-Pick C1

Infections by the Ebola and Marburg filoviruses cause a rapidly fatal haemorrhagic fever in humans for which no approved antivirals are available. Filovirus entry is mediated by the viral spike glycoprotein (GP), which attaches viral particles to the cell surface, delivers them to endosomes and catalyses fusion between viral and endosomal membranes. Additional host factors in the endosomal compartment are probably required for viral membrane fusion; however, despite considerable efforts, these critical host factors have defied molecular identification. Here we describe a genome-wide haploid genetic screen in human cells to identify host factors required for Ebola virus entry. Our screen uncovered 67 mutations disrupting all six members of the homotypic fusion and vacuole protein-sorting (HOPS) multisubunit tethering complex, which is involved in the fusion of endosomes to lysosomes, and 39 independent mutations that disrupt the endo/lysosomal cholesterol transporter protein Niemann-Pick C1 (NPC1). Cells defective for the HOPS complex or NPC1 function, including primary fibroblasts derived from human Niemann-Pick type C1 disease patients, are resistant to infection by Ebola virus and Marburg virus, but remain fully susceptible to a suite of unrelated viruses. We show that membrane fusion mediated by filovirus glycoproteins and viral escape from the vesicular compartment require the NPC1 protein, independent of its known function in cholesterol transport. Our findings uncover unique features of the entry pathway used by filoviruses and indicate potential antiviral strategies to combat these deadly agents.     

埃博拉病毒感染的实验室诊断方法研究进展

 埃博拉病毒病是由埃博拉病毒引起的一种急性出血性传染病,具有极高的传染性,病死率高达90%,世界卫生组织已将埃博拉病毒列为对人类危害最严重的病毒之一,目前虽有一个被美国食品和药物管理局(FDA)批准用于紧急治疗的实验药物TKM-Ebola,但尚无批准上市的用于预防埃博拉病毒的疫苗.建立快速、准确、简便的实验室检测方法对及时进行临床诊断救治、开展流行病学调查并最终控制其传播流行具有重要意义.本文综述埃博拉病毒实验室检测方法的原理、应用及进展,介绍病毒分离、电子显微镜观察、逆转录聚合酶链反应法、抗原检测试验、抗体酶联免疫吸附试验、血清中和试验及其他检测方法.     

Small molecule inhibitors reveal Niemann-Pick C1 is essential for Ebola virus infection

Ebola virus (EboV) is a highly pathogenic enveloped virus that causes outbreaks of zoonotic infection in Africa. The clinical symptoms are manifestations of the massive production of pro-inflammatory cytokines in response to infection and in many outbreaks, mortality exceeds 75%. The unpredictable onset, ease of transmission, rapid progression of disease, high mortality and lack of effective vaccine or therapy have created a high level of public concern about EboV. Here we report the identification of a novel benzylpiperazine adamantane diamide-derived compound that inhibits EboV infection. Using mutant cell lines and informative derivatives of the lead compound, we show that the target of the inhibitor is the endosomal membrane protein Niemann-Pick C1 (NPC1). We find that NPC1 is essential for infection, that it binds to the virus glycoprotein (GP), and that antiviral compounds interfere with GP binding to NPC1. Combined with the results of previous studies of GP structure and function, our findings support a model of EboV infection in which cleavage of the GP1 subunit by endosomal cathepsin proteases removes heavily glycosylated domains to expose the amino-terminal domain, which is a ligand for NPC1 and regulates membrane fusion by the GP2 subunit. Thus, NPC1 is essential for EboV entry and a target for antiviral therapy.     

普通棉耳狨猴中乙型流感病毒的分离和鉴定

乙型流感病毒是引起人类局部流行性感冒的重要病原体，其起源和自然储存宿主目前仍不清楚。1999年夏季在某动物中心饲养的普通棉耳绒猴（Callithrix jacchus）群体中爆发了以呼吸道症状为主的急性传染病，死亡比率高达1／3。通过对死亡狨猴肺组织匀浆接种鸡胚和MDCK细胞的分离培养，分离出病毒。双份血清的红细胞凝集抑制试验证实，此分离株为此次狨猴群体感染流行的病原体。通过甲型和乙型流感病毒标准血清鉴定，证实该分离株为乙型流感病毒，命名为B/marmoset/China/1/99。     

Spontaneous and continuous anti-virus disinfection from nonstoichiometric perovskite-type lanthanum manganese oxide

Viral pathogens have threatened human being's health for a long time, from periodically breakout flu epidemics to recent rising Ebola virus disease. Herein, we report a new application of nonstoichiometric Perovskite-type LaxMn O3(x ? 1, 0.95, and 0.9) compounds in spontaneous and continuous disinfection of viruses. Perovskite-type LaxMn O3(x ? 1, 0.95, and 0.9) is well-known for their catalytic properties involving oxidization reactions, which are usually utilized as electrodes in fuel cells. By utilizing superb oxidative ability of LaxMn O3(x ? 1, 0.95, and 0.9),amino acid residues in viral envelope proteins are oxidized, thus envelope proteins are denatured and infectivity of the virus is neutralized. It is of great importance that this process does not require external energy sources like light or heat. The A/PR/8/34H1N1 influenza A virus(PR8) was employed as the sample virus in our demonstration, and high-throughput disinfections were observed. The efficiency of disinfection was correlated to oxidative ability of LaxMn O3(x ? 1, 0.95, and 0.9) by EPR and H2-TPR results that La0.9Mn O3 had the highest oxidative ability and correspondingly gave out the best disinfecting results within three nonstoichiometric compounds. Moreover, denaturation of hemagglutinin and neuraminidase, the two key envelope proteins of influenza A viruses, was demonstrated by HA unit assay with chicken red blood cells and NA fluorescence assay, respectively. This unique disinfecting application of La0.9Mn O3 is considered as a great make up to current sterilizing methods especially to photocatalyst based disinfectants and can be widely applied to cut-off spread routes of viruses, either viral aerosol or contaminated fluid, and help in controlling the possibly upcoming epidemics like flus and hemorrhagic fever.     

Human immunodeficiency viruses: SIV infection in wild gorillas

Chimpanzees (Pan troglodytes troglodytes) from west central Africa are recognized as the reservoir of simian immunodeficiency viruses (SIVcpzPtt) that have crossed at least twice to humans: this resulted in the AIDS pandemic (from human immunodeficiency virus HIV-1 group M) in one instance and infection of just a few individuals in Cameroon (by HIV-1 group N) in another. A third HIV-1 lineage (group O) from west central Africa also falls within the SIVcpzPtt radiation, but the primate reservoir of this virus has not been identified. Here we report the discovery of HIV-1 group O-like viruses in wild gorillas.     

Structure of the Ebola virus glycoprotein bound to an antibody from a human survivor

Ebola virus (EBOV) entry requires the surface glycoprotein (GP) to initiate attachment and fusion of viral and host membranes. Here we report the crystal structure of EBOV GP in its trimeric, pre-fusion conformation (GP1+GP2) bound to a neutralizing antibody, KZ52, derived from a human survivor of the 1995 Kikwit outbreak. Three GP1 viral attachment subunits assemble to form a chalice, cradled by the GP2 fusion subunits, while a novel glycan cap and projected mucin-like domain restrict access to the conserved receptor-binding site sequestered in the chalice bowl. The glycocalyx surrounding GP is likely central to immune evasion and may explain why survivors have insignificant neutralizing antibody titres. KZ52 recognizes a protein epitope at the chalice base where it clamps several regions of the pre-fusion GP2 to the amino terminus of GP1. This structure provides a template for unravelling the mechanism of EBOV GP-mediated fusion and for future immunotherapeutic development.     

Global trends in emerging infectious diseases

Emerging infectious diseases (EIDs) are a significant burden on global economies and public health. Their emergence is thought to be driven largely by socio-economic, environmental and ecological factors, but no comparative study has explicitly analysed these linkages to understand global temporal and spatial patterns of EIDs. Here we analyse a database of 335 EID 'events' (origins of EIDs) between 1940 and 2004, and demonstrate non-random global patterns. EID events have risen significantly over time after controlling for reporting bias, with their peak incidence (in the 1980s) concomitant with the HIV pandemic. EID events are dominated by zoonoses (60.3% of EIDs): the majority of these (71.8%) originate in wildlife (for example, severe acute respiratory virus, Ebola virus), and are increasing significantly over time. We find that 54.3% of EID events are caused by bacteria or rickettsia, reflecting a large number of drug-resistant microbes in our database. Our results confirm that EID origins are significantly correlated with socio-economic, environmental and ecological factors, and provide a basis for identifying regions where new EIDs are most likely to originate (emerging disease 'hotspots'). They also reveal a substantial risk of wildlife zoonotic and vector-borne EIDs originating at lower latitudes where reporting effort is low. We conclude that global resources to counter disease emergence are poorly allocated, with the majority of the scientific and surveillance effort focused on countries from where the next important EID is least likely to originate.     

Administration of vorinostat disrupts HIV-1 latency in patients on antiretroviral therapy

Despite antiretroviral therapy, proviral latency of human immunodeficiency virus type 1 (HIV-1) remains a principal obstacle to curing the infection. Inducing the expression of latent genomes within resting CD4(+) T cells is the primary strategy to clear this reservoir. Although histone deacetylase inhibitors such as suberoylanilide hydroxamic acid (also known as vorinostat, VOR) can disrupt HIV-1 latency in vitro, the utility of this approach has never been directly proven in a translational clinical study of HIV-infected patients. Here we isolated the circulating resting CD4(+) T cells of patients in whom viraemia was fully suppressed by antiretroviral therapy, and directly studied the effect of VOR on this latent reservoir. In each of eight patients, a single dose of VOR increased both biomarkers of cellular acetylation, and simultaneously induced an increase in HIV RNA expression in resting CD4(+) cells (mean increase, 4.8-fold). This demonstrates that a molecular mechanism known to enforce HIV latency can be therapeutically targeted in humans, provides proof-of-concept for histone deacetylase inhibitors as a therapeutic class, and defines a precise approach to test novel strategies to attack and eradicate latent HIV infection directly.     

Catastrophic ape decline in western equatorial Africa

Because rapidly expanding human populations have devastated gorilla (Gorilla gorilla) and common chimpanzee (Pan troglodytes) habitats in East and West Africa, the relatively intact forests of western equatorial Africa have been viewed as the last stronghold of African apes. Gabon and the Republic of Congo alone are thought to hold roughly 80% of the world's gorillas and most of the common chimpanzees. Here we present survey results conservatively indicating that ape populations in Gabon declined by more than half between 1983 and 2000. The primary cause of the decline in ape numbers during this period was commercial hunting, facilitated by the rapid expansion of mechanized logging. Furthermore, Ebola haemorrhagic fever is currently spreading through ape populations in Gabon and Congo and now rivals hunting as a threat to apes. Gorillas and common chimpanzees should be elevated immediately to 'critically endangered' status. Without aggressive investments in law enforcement, protected area management and Ebola prevention, the next decade will see our closest relatives pushed to the brink of extinction.     

野生鸟类在禽流感传播中所起的作用

野生鸟类很可能是高致病性禽流感（HPAI）在全球传播的媒介．这一论点来自于全球机构在过去数十年对野生鸟类体内禽流感病毒的调查、检测和统计结果．这一结果表明：（1）禽流感病毒能够感染所有种类的野鸟,但以雁鸭类为主的水禽是病毒的天然贮存库;（2）多数禽流感病毒亚型对宿主具有选择性,共同感染多种病毒亚型具有非随机性,这使病毒基因重组突变产生亚型的机会并不多,此结论建议防控野生鸟类传播禽流感的基本措施是：（1）隔离各类动物,避免相互接触．提高生物安全措施;（2）针对数量庞大的水禽．建立高效的检测和监测机制．     

新场上沙溪庙致密碎屑岩气田整体压裂开发方案设计研究

为了解决川西致密碎屑岩气田存在的生产动态复杂、储量难动用、采速低、见产慢、稳产条件差、提高综合开采效益困难等问题和难点,通过研究和现场试验,认为采用整体压裂改造开发技术可以获得较好的产能和经济开发效果。建立了气、水三维二相三重介质低渗致密裂缝性气藏整体压裂模拟模型,模型中首次全面考虑了低渗致密气藏低速非达西型渗流和高速非达西型渗流的影响,结合地应力研究,对新场上沙溪庙致密碎屑岩气田整体压裂开发方案进行了设计研究;为低渗致密气藏整体压裂改造方案的编制提供了思路和工具。     

川东北宣汉-达县地区储层预测技术

川东北宣汉-达县地区以构造圈闭为主要勘探对象,钻探成功率低,为此开展了以储层预测技术为主导的构造-岩性复合圈闭勘探.通过山区地震数据高分辨率采集、处理、储层沉积相研究、储层测井响应特征、储层物性特征及储层层位标定、地震相研究、储层地震预测方法研究,确立了研究区内的三大储层类型及各类储层的研究方法,提出了川东北复杂储层预测以野外地震资料高分辨率采集为基础、多种特殊处理手段相结合,并以沉积相研究为指导的综合性研究思路,为川东北宣汉-达县地区储层预测方法研究提供了指导性意见,为该地区复杂储层勘探的深入与突破打了坚实的基础.     

基于岩石物理相研究特低渗透油藏宏观剩余油分布

对于低渗、特低渗透储层,由于其所受沉积、成岩和构造等一系列作用的影响,致使该类油藏的储集性能和渗流结构存在较大的差异,储层具有较强的非均质性,剩余油分布异常复杂;而储层的非均质性在一定程度上控制着该类油藏的剩余油分布。因此,以延长东部某特低渗透非均质油藏长6_1~1油层为例,提出利用岩石物理相研究该类非均质油藏宏观剩余油的方法。通过建立岩石物理相定量综合评价指标体系,将储集层分为几种不同类别的岩石物理相。针对不同类别岩石物理相分别计算单井储量、标定技术采收率以及研究剩余可采储量分布。研究结果表明岩石物理相是控制该类油藏剩余油分布的基本单元,同种岩石物理相内剩余油的分布规律相似,从而阐明了有利的岩石物理相剩余油潜力分布的特点,实现了宏观剩余油的研究从笼统研究到精细化研究的转化,为油区后期剩余油的挖潜提供了可靠的地质依据。同时也为同类型的特低渗透非均质油藏剩余油潜力分布的研究提供了新颖的研究思路和方法。     

The genomic and epidemiological dynamics of human influenza A virus

The evolutionary interaction between influenza A virus and the human immune system, manifest as 'antigenic drift' of the viral haemagglutinin, is one of the best described patterns in molecular evolution. However, little is known about the genome-scale evolutionary dynamics of this pathogen. Similarly, how genomic processes relate to global influenza epidemiology, in which the A/H3N2 and A/H1N1 subtypes co-circulate, is poorly understood. Here through an analysis of 1,302 complete viral genomes sampled from temperate populations in both hemispheres, we show that the genomic evolution of influenza A virus is characterized by a complex interplay between frequent reassortment and periodic selective sweeps. The A/H3N2 and A/H1N1 subtypes exhibit different evolutionary dynamics, with diverse lineages circulating in A/H1N1, indicative of weaker antigenic drift. These results suggest a sink-source model of viral ecology in which new lineages are seeded from a persistent influenza reservoir, which we hypothesize to be located in the tropics, to sink populations in temperate regions.