EV71疫苗研究进展

肠道病毒71型(enterovims 71,EV71)是手足口病的主要病原体,由于尚无治疗手足口病的有效抗病毒药,因此,开发预防手足口病的有效EV71疫苗成为所有控制措施的首选.目前,EV71疫苗研究已取得一定的进展,正在研发的疫苗包括减毒或灭活疫苗、VP1亚单位疫苗和DNA疫苗.虽然各种疫苗均具有一定的免疫效果,但最适疫苗的确认还需对各种疫苗的安全性和保护效力进行评估.     

EV71疫苗研究进展

肠道病毒71型(enterovims 71,EV71)是手足口病的主要病原体,由于尚无治疗手足口病的有效抗病毒药,因此,开发预防手足口病的有效EV71疫苗成为所有控制措施的首选.目前,EV71疫苗研究已取得一定的进展,正在研发的疫苗包括减毒或灭活疫苗、VP1亚单位疫苗和DNA疫苗.虽然各种疫苗均具有一定的免疫效果,但最适疫苗的确认还需对各种疫苗的安全性和保护效力进行评估.     

我国柯萨奇病毒A组16型疫苗的研究进展

 手足口病是一种主要由柯萨奇病毒A组16型（coxsackievirus A16,CVA16）和肠道病毒71型（enterovirus 71,EV71）引起的全球性传染病,婴幼儿最容易受病毒感染。疫苗接种是控制手足口病的主要措施。与EV71疫苗相比,CVA16疫苗的研究相对滞后,但EV71疫苗的顺利研发促进了CVA16疫苗的研究。此文就我国CVA16灭活疫苗、病毒样颗粒疫苗、亚单位疫苗、DNA疫苗以及CVA16与 EV71联合疫苗的研究进展做一综述。     

Review of hand, foot and mouth disease

Hand, foot and mouth disease (HFMD) is an acute viral illness that primarily affects infants and young children, and often occurs in clusters or outbreaks. The major causative agents of HFMD are coxsackievirus A16 (CVA16), human enterovirus 71 (HEV71) and coxsackievirus A10 (CVA10), of the genus Enterovirus in the family Picornaviridae. HFMD caused by EV71 is associated with severe neurological complications and death. Since the late 1990s, several major epidemics of EV71 HFMD have swept through the Asia-Pacific region, associated with a rapid fulminant course, severe neurological complications, and a large number of fatalities. Until now, little has been known about the genetics and transmission trends of the fast-mutation virus, and there is no effective and specific antiviral therapy or vaccine for HFMD. It is time to step up efforts to understand the molecular epidemics and pathogenesis so that we can develop effective management strategies and surveillance programs.     

Prevention of human enterovirus 71 infection by kappa carrageenan

Enterovirus 71 (EV 71), the newest member of Enteroviridae, is notorious for its etiological role in epidemics of the hand-foot-and-mouth disease, particularly in association with fatal neurological complications in young children. Searching for new and more effective agents against EV 71 infections has never relented as corresponding vaccines or antiviral drugs remain unavailable. Sulfated polysaccharides from seaweed are known to possess a broad range of biological activities across anti-virus, anti-tumor, immunomodulation, anti-coagulation, etc. In this study, we report kappa carrageenan also has a strong and effective anti-EV 71 activity able to reduce plaque formation, prevent viral replication before or during viral adsorption, as well as inhibit EV 71-induced apoptosis. In virus binding assay, kappa carrageenan was shown able to bind EV 71 firmly, forming carrageenan-viruses complexes, whereby the virus-receptor interaction is likely disrupted. Added together, kappa carrageenan may be an ideal candidate worthwhile to develop into anti-EV 71 agents.     

Measures for control of influenza

Influenza viruses cause recurring illnesses among individuals and recurring epidemics among populations. The major effective control measure for preventing infection and illness is inactivated vaccine, which can prevent influenza illnesses and their complications when given before exposure to the virus. While inactivated vaccine is effective for preventing influenza in most individuals, recommendations for its use focus on the prevention of severe disease and death among those who are at high risk of complications. Live attenuated cold-adapted influenza vaccines are nearing availability. They are given by nasal spray and are particularly effective for preventing influenza among young children, but also for preventing influenza among young adults, and enhancing protection against influenza when given with inactivated vaccine to elderly persons. The antiviral agents amantadine and rimantadine are related compounds that are effective for the prevention and treatment of influenza A virus infections and illnesses. Disadvantages are the rapid development of resistance during treatment and CNS adverse effects with amantadine. These drugs are also effective for outbreak control. Ribavirin is an antiviral given by small particle aerosol that is approved for the treatment of respiratory syncytial virus disease; it is also effective for the treatment of influenza. Two new antiviral agents inhibit influenza viral neuraminidase activity; one is given by inhalation or intranasally (zanamivir) and the other orally (GS4104). The former is free of adverse effects, while the latter induces nausea and vomiting in some individuals. Both are effective for the prevention as well as the treatment of influenza A and B illnesses. Thus, various measures for preventing and treating influenza are nearing availability. Their optimal use should further improve the control of influenza in individuals and populations as well as permit efforts to prevent community epidemics.     

肠道病毒71型疫苗的研究进展

肠道病毒71型(enterovirus 71,EV71)是近年来我国流行的手足口病的主要病原体之一,并且由其感染引发的重症和死亡病例的比例较大.目前尚无治疗手足口病的特效药物.我国大陆有3家单位完成了EV71灭活疫苗的Ⅲ期临床试验,国家食品药品监督管理总局批准了其中两家单位的灭活疫苗.目前,减毒活疫苗、病毒样颗粒疫苗、亚单位疫苗和DNA疫苗等已开展动物研究.     

我国肠道病毒71型（EV71）疫苗株生物信息学分析

目的 比较3个EV71疫苗株病毒的生物信息学特点,为疫苗的质控和探讨该病毒疫苗的免疫机制提供参考.方法 本研究利用DNAstar MegAlign、DNAMAN Alignment、ANtheProt等生物学软件,比较了我国已进入临床试验的3家EV71全病毒灭活疫苗毒株的基因组及氨基酸序列,并对英衣壳蛋白二级结构及可能存在的抗原表位进行了预测分析.结果 3个疫苗株病毒基因同源性为97％～99％;氨基酸同源性为98％～99％;二级结构一致率在95％以上;均存在35个潜在抗原表位区域.结论 3个疫苗株生物信息学分析结果存在高度的一致性,提示疫苗具有相似的抗原性和免疫原性.     

肠道病毒71型病毒样颗粒疫苗研究进展

肠道病毒71型(enterovirus 71,EV71)是引起儿童手足口病的主要病原体之一.由于尚无有效的抗病毒药物,故研发安全有效的疫苗是控制手足口病的有效措施.目前研发的EV71疫苗以灭活疫苗和病毒样颗粒(virus-like particle,VLP)疫苗为主,研究表明,VLP具有良好的免疫原性和稳定性.此文对目前EV71 VLP疫苗的研究进展做一综述,以期为预防EV71相关手足口病及其他手足口病的新型疫苗的研制提供思路.     

肠道病毒71型疫苗研究进展及质量标准

肠道病毒71型(enterovirus 71,EV71)是导致手足口病的主要病原体之一,近年来在我国持续流行,对婴幼儿的健康造成了严重威胁.安全有效的疫苗是控制EV71流行的重要手段.全球有多家研究机构正在开展EV71疫苗的相关研究,目前由我国自主研发的3种EV71疫苗已获准上市.同时,为保证疫苗的安全性、有效性和质量可控性,已在研发中建立了合适且规范的质量标准.     

肠道病毒71型的研究进展

肠道病毒71型(enterovirus 71,EV71)是引起手足口病(hand,foot and mouth disease,HFMD)的主要病原体之一,HFMD可并发严重神经系统疾病,甚至导致死亡.事实上,EV71一直被视为继脊髓灰质炎病毒消灭后最重要的嗜神经病毒.此文就EV71感染的易感因素以及疫苗的研究进展进行综述,从而为EV71感染的防控指明方向.     

肠道病毒71型的研究进展

肠道病毒71型(enterovirus 71,EV71)是引起手足口病(hand,foot and mouth disease,HFMD)的主要病原体之一,HFMD可并发严重神经系统疾病,甚至导致死亡.事实上,EV71一直被视为继脊髓灰质炎病毒消灭后最重要的嗜神经病毒.此文就EV71感染的易感因素以及疫苗的研究进展进行综述,从而为EV71感染的防控指明方向.     

磁珠法结合定量PCR检测肠道病毒71型灭活疫苗中宿主细胞DNA残留量的验证及应用

目的  对磁珠法结合定量PCR（quantitative PCR,qPCR）检测肠道病毒71型灭活疫苗〔非洲绿猴肾细胞（Vero细胞）〕（EV71疫苗）中宿主DNA残留量进行适用性验证及应用。方法  利用微磁珠与蛋白溶液中DNA的特异性结合,来提取样品中残留的Vero细胞DNA。将已知浓度Vero细胞DNA对照系列稀释后,作为标准品DNA与提取的DNA同时进行qPCR扩增。根据标准品DNA的循环阈值与浓度之间的线性关系,对未知样品中残留DNA进行定量分析。结果  标准品检测范围在0.03～3 000.00 pg/反应。该方法的标准曲线决定系数≥0.98,扩增效率在90%～110%。质控样品回收率在50%～150%,相对标准偏差均小于30%。实验结果的各项参数均在要求范围内。结论  磁珠法可解决残留DNA检测中样品前处理的技术难点,qPCR能够简便、快速、准确地对生产过程样品的EV71疫苗中DNA残留量进行定量测定。适用于EV71疫苗中DNA残留量的检测及疫苗生产过程和其成品的质量控制,对其他以Vero细胞为基质的病毒性疫苗质量控制具有借鉴意义。     

复方一枝蒿颗粒对防治手足口病的体内外药效学研究

目的 通过观察复方一枝蒿颗粒在体内外对肠道病毒的抑制作用,评价复方一枝蒿颗粒防治手足口病的药效作用。方法 采用复方一枝蒿颗粒的3岁儿童临床2倍、等倍和1/2倍剂量分别对肠道病毒EV71 H株感染的Babl/c乳鼠手足口病动物模型进行预防性给药和治疗性给药,通过观察乳鼠感染的严重程度、死亡数,来计算动物死亡率、死亡保护率和生命延长率;采用细胞病变的CPE法观察复方一枝蒿颗粒对肠道病毒EV71 H株,BrCr株,柯萨奇病毒B3、B4、B5株的抑制作用;采用real time PCR法观察复方一枝蒿颗粒对肠道病毒EV71 BrCr株在RD细胞中增殖的抑制作用。结果 复方一枝蒿颗粒对肠道病毒EV71 H株感染的乳鼠模型具有明显的治疗作用,其可降低感染程度和死亡率,具有明显的死亡保护和延长生命的作用;复方一枝蒿颗粒在无明显毒性的浓度下,对肠道病毒EV71 H株、BrCr株具有明显的抑制作用,其能明显抑制EV71 BrCr株在细胞内的增殖。结论 复方一枝蒿颗粒在体内外对手足口病均具有较好的治疗作用,其作用机制可能与抑制肠道病毒在细胞内增殖有关。     

EV71非结构蛋白的结构和功能及其为靶点的药物研究进展

近年肠道病毒71型(EV71)等引起的手足口病在中国大陆呈现上升的流行趋势,其正链的RNA基因组翻译成一个多聚蛋白,进一步自剪切为结构蛋白和非结构蛋白;随着研究的深入,非结构蛋白在病毒生命周期中的功能逐一被鉴定,本文就EV71非结构蛋白的结构功能及针对这些靶点的抗病毒药物进行综述。     

肠道病毒71型抗原定量检测方法的建立

目的 建立肠道病毒71型(enterovirus 71,EV71)抗原的定量检测方法,用于EV71疫苗生产过程中EV71抗原含量的监测.方法 用EV71抗原免疫新西兰兔制备抗EV71多克隆抗体,纯化后用辣根过氧化物酶标记.采用双抗体夹心ELISA法建立抗原检测系统.以EV71国家抗原标准品为定量标准,建立剂量反应曲线,确定该方法的线性范围和定量限,并对该方法的特异性、精密度、准确性、稳定性及适用性等进行验证.结果 建立的ELISA法的线性范围为5～80 U/ml,定量限为5 U/ml,线性决定系数均大于0.990 0;不同浓度样品回收率为85.0％～110.0％,变异系数均小于15％;置于2～8℃及37℃3、6d的抗体包被板对不同浓度样品的检测值变异系数均小于15％;该方法可特异性检测EV71原液,与非EV71样品无交叉反应.结论 建立了EV71抗原定量检测方法,为EV71疫苗生产工艺过程的质量控制奠定了基础.     

Fighting against infectious diseases in China

Dr. Yin started his research on infectious disease prevention in the 1980s. In 1985, Dr. Yin sucessfully isolated the hepatitis A virus, after which, in 2002, he developed the first proprietary inactivated hepatitis A vaccine in China and soon launched it into the China market. Led by Dr. Yin, Sinovac successfully developed the vaccine prducts against SARS, H5N1, H1N1, hepatitis A and B and infleunza. Currently, Sinovac is working on the R&D of EV71 vaccine against hand, foot and mouth disease, and pneumococcal conjugate vaccine. Sinovac aims to provide Chinese children with international quality vaccines, and provide children in the world with vaccines made in China.     

肠道病毒71型与宿主相互作用

肠道病毒71型(enterovirus 71,EV71)是引起手足口病的主要病原体之一.EV71传播广泛,感染后可引发中枢神经系统疾病,并导致重症手足口病,给公共卫生安全带来极大挑战.EV71的致病机制与病毒和宿主间的相互作用关系密切,涉及EV71病毒复制、微RNA等多个环节.此文就近年来这些方面的研究进展做一综述.     

Induction of protective immune responses against EV71 in mice by baculovirus encoding a novel expression cassette for capsid protein VP1

EV71 is a major causative agent of hand, foot and mouth disease (HFMD) and is responsible for large outbreaks in various Asian Pacific countries. In the present study, we generated the recombinant baculovirus (Bac-VP1) encoding VP1 in a novel expression cassette. The transmembrane domain of hemagglutinin of the H3N2 influenza virus was included in the cassette as a minimal membrane anchor for VP1. The protective immunity of Bac-VP1 was investigated in a mouse model. The results showed that mice vaccinated with live Bac-VP1 had strong VP1 specific antibody responses. In an in vitro neutralization assay Bac-VP1 sera exhibited cross-neutralization against homologous and heterologous EV71 strains with a maximum titer of 1:512. Passive immunization studies confirmed that these sera were able to provide 100% protection against 5 MLD(50) of mouse adapted EV71 (B4 strain). This study revealed that baculovirus displaying VP1 with a HA transmembrane domain efficiently induced cross-neutralizing antibody responses in mice.     

Vero细胞乙型脑炎灭活疫苗的研制

目的 以Vero细胞为基质,研制安全有效的乙型脑炎灭活疫苗.方法 将经Vero细胞适应的乙型脑炎病毒P3株接种Vero细胞,培养后收获病毒液.采用β-丙内酯灭活,通过超滤浓缩、硫酸鱼精蛋白沉淀、蔗糖密度梯度离心等方法纯化病毒,最终配制成预防乙型脑炎病毒感染的疫苗.通过临床试验考察疫苗的安全性及免疫原性.结果 该疫苗的主要考核指标,如病毒滴度、总蛋白含量、Vero细胞DNA残留量、效价等均符合国家标准.临床观察结果显示,接种本疫苗后局部及全身反应轻微;各年龄组乙型脑炎病毒中和抗体阳转率均>90%.结论 采用本法制备的疫苗安全有效.