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手足口病在世界多个地区,尤其是亚洲爆发并流行,且其感染率和死亡率逐年增高,危害十分严重。肠道病毒71(Enterovirus 71,EV71)是手足口病(Hand,foot,andmouth disease,HFMD)的主要病原体,以感染婴幼儿为主,其感染常伴随神经系统并发症,严重可导致儿童死亡。近年来,分子生物学和抗病毒研究方面取得的进展为EV71感染的预防及治疗提供了新的途径。本文对EV71病毒学特点及抗EV71药物的筛选、疫苗开发、RNA干扰等进行了综述,以期为相关研究提供参考。 相似文献
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肠道病毒A71型(Enterovirus A71,EV-A71)可引起手足口病(Hand,foot,and mouth disease,HFMD),严重者伴有神经系统并发症,如无菌性脑膜炎、神经源性肺水肿等。EV-A71引起的HFMD自2007年以来在全世界,尤其是亚太地区多次暴发,已成为亚太地区公共健康的主要威胁之一。目前尚无有效的抗病毒药物或疫苗。EVA71的致病机制尚未完全研究清楚,而非结构蛋白3C在病毒的复制和抑制天然免疫方面发挥了不可替代的作用。EV-A71 3C蛋白的研究在进一步了解EV-A71的致病机制以及研制抗病毒药物方面发挥着重要的作用。本文将对EV-A71 3C蛋白的结构、功能以及抗3C蛋白病毒药物的研究进展做出综述。 相似文献
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肠道病毒71型（Enterovirus 71,EV71）是引起重症手足口病（Hand,Foot and Mouth Disease,HFMD）的主要病原体。重症HFMD进展迅速,可表现为严重的神经系统并发症,甚至危及生命。目前临床上防治EV71感染缺乏特异、高效的药物,其残疾率和死亡率很高。随着研究的深入,已经发现了大量具有抗EV71能力的化合物,人们探索的药物机制和药物靶点各不相同。因此,本文从药物靶向病毒、宿主等角度出发,针对抗EV71感染的天然药物、合成药物及常见中药中活性成分作用机制的最新进展进行综述与讨论。此外,对抗病毒药物筛选技术进行简要概述,以期为抗EV71药物的筛选与研发设计等相关研究提供参考。 相似文献
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肠道病毒71型(Enterovirus 71,EV71)为小RNA病毒科肠道病毒属A组病毒的代表株,感染可引发手足口病(Hand-foot and mouth disease,HFMD),严重危害儿童健康.EV71 3C蛋白酶(3C)是其编码的主要蛋白酶之一,在病毒多蛋白加工过程中发挥关键作用,同时切割细胞蛋白以利于病毒复制.为进一步了解EV71与宿主间博弈关系,本课题组前期以3C为诱饵进行酵母双杂交实验,筛选与3C发生相互作用的可能底物,钓取到锌指MYM型蛋白2(Zinc finger MYM-type protein 2,ZMYM2).ZMYM2是一种具有锌指结构的转录因子,与细胞中重要的抗病毒小体PML核体(PML nuclear bodies,PML-NBs)的形成及稳定性相关.本文选择3C与ZMYM2关系展开研究,确证二者相互作用并初探生物学功能.首先通过免疫共沉淀实验确证3C与ZMYM2之间存在相互作用;随后分析功能,发现过表达ZMYM2抑制EV71复制;敲减内源ZMYM2有利于EV71的复制;分析3C对ZMYM2影响,发现3C剂量依赖性切割ZMYM2,ZMYM2上至少具有2个3C的识别位点.本研究为解析ZMYM2功能及进一步了解EV71与宿主先天免疫间博弈关系提供了新的实验证据. 相似文献
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目的:建立以蛋白酶Neprilysin(NEP)为靶点的高通量药物筛选模型,应用该模型筛选抑制剂。方法:利用毕赤酵母表达系统。构建重组质粒p PICZα-A-NEP,表达载体通过与酵母菌X-33基因组染色体发生同源重组,将外源基因整合于染色体后实现目的蛋白的表达。应用荧光共振能量转移法(FRET)检测蛋白酶活性,优化反应条件,建立药物筛选体系,筛选抑制剂。结果:成功构建表达载体p PICZα-A-NEP;建立了以NEP为靶标的药物筛选模型,获得模型反应动力学参数Vmax=3.6μM/s,Kcat/Km=4.5×105M-1s-1,测定模型Z-因子为0.89,说明体系稳定可用于以NEP为靶标的药物的高通量筛选;并用该模型对天然产物组分库进行筛选,在0.5mg/ml的药物浓度下,得到抑制率较高的药物为4种,并测得半数抑制浓度IC50值,其中MDCNCL01000242的IC50值最低,为(8.31±0.03)μg/ml。结论:建立的药物筛选模型较为理想,适用于NEP抑制剂的筛选,可促进药物的研发。 相似文献
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为研究肠道病毒71型(Enterovirus 71,EV-A71)经不同攻毒方式感染不同日龄ICR乳鼠的感染情况,了解EV-A71在小鼠体内的动态分布和感染机制,为建立EV-A71感染动物模型,本研究采用分离自重症手足口患儿的EV-A71毒株,分别通过肌肉注射(Intramuscular injection,IM)、腹腔注射(Intraperitoneal injection,IP)以及脑内注射(Intracerebral injection,IC)的方式感染3日、5日和9日龄ICR乳鼠,感染后定期采集血液和各组织,通过Realtime PCR追踪各组织中病毒载量变化,并且通过切片制作和免疫组化对感染乳鼠进行病原学和病理学分析。结果显示:对于低日龄(≤5日龄)乳鼠,注射剂量在104.5 TCID50/g·体重时IM、IP及IC均为较好的感染方式,都会出现神经症状,且有极高的致死率。随着日龄增长,感染后症状有所减轻,但IM和IP感染途径对大日龄乳鼠仍具有良好的致病性,且IM和IP死亡率显著高于IC死亡率。经IM、IP和IC注射感染病毒的3日龄乳鼠在6dpi体重相对于各对照组分别下降了1.54g(31.43%)、1.31g(15.06%)和2.52g(44.28%),而经IM和IP感染的5日龄乳鼠6dpi体重相对于各对照组分别下降了0.605g(8.95%)、0.886g(15.51%),经IC感染的5日龄乳鼠6dpi体重相对于对照组上升了0.904g(14.70%),感染组体重均显著低于对应的同期对照组(P0.05)。经IM、IP和IC感染病毒的3日龄乳鼠9dpi均死亡,而5日龄乳鼠9dpi存活率分别为42.8%、25%和87.5,14dpi存活率分别为0%、0%和25%,9日龄乳鼠9dpi存活率分别为70%、84.62%和100%。病理学及免疫组化检查显示EV-A71病毒具有强烈的嗜神经性及嗜骨骼肌的特性,可导致病毒血症、脑神经元及骨骼肌坏死、心肌间质水肿及多脏器炎症反应。我们系统地研究了不同攻毒方式感染不同日龄的ICR乳鼠后的病毒动态分布及免疫病理损伤,发现经肌肉注射或腹腔注射5日龄乳鼠能够建立理想的EV-A71感染动物模型。 相似文献
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对人肠道病毒EV71型2009年广州分离株Guangzhou09构建序列系统进化树并分析其重组特点。P1、P2和P3区种系发生进化树提示该分离株发生重组,相似性曲线以及bootscan进一步分析显示该分离株于非结构编码蛋白2B片段区(核苷酸位置4 027bp)处存在EV71亚型C4Shanghai-FJ713137与CVA409-HQ728260型间重组。该分离株与中国大陆目前优势株流行趋势一致,是广州首例发生型间重组且由C4亚型及CVA4型发生重组。 相似文献
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根据嗜杀酵母T158c/S14a中L-A病毒-1移码效率改变影响M1病毒的存活,导致K1毒素减少,在低pH的美蓝平板上用杯碟法通过抑菌圈的大小检测酵母K1毒素的嗜杀活性,建立了一个以酵母嗜杀系统为基础的抗病毒药物筛选模型。研究了杯碟法检测酵母毒素嗜杀活性的各种条件。对不同pH和温度下酵母的嗜杀活性进行了研究,确定了模型用于筛选的最适pH范围为4.3~4.7,最适温度范围为20~22℃。运用该模型研究了几种中药对嗜杀活性的抑制作用,发现了金银花和升麻具有一定的抗病毒作用。该模型为抗病毒药物的高通量初筛奠定了基础。 相似文献
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Chen Zhao Liuyun Tang Haixia Ren Peng Li Ning Li Jianwei Huang Xueqin Chen Yi Guan Han You Shuhui Chen Jian Li Tianwei Lin 《Journal of molecular recognition : JMR》2016,29(11):520-527
Enterovirus 71 (EV71) is the causative agent of hand, foot and mouth disease and can spread its infections to the central nervous and other systems with severe consequences. The replication of EV71 depends on its 3C proteinase (3Cpro), a significant drug target. By X‐ray crystallography and functional assays, the interactions between inhibitors and EV71 3Cpro were evaluated. It was shown that improved interactions at S4 for the substrate binding could significantly enhance the potency. A new series of potent inhibitors with high ligand efficiency was generated for developing antivirals to treat and control the EV71‐associated diseases. Copyright © 2016 John Wiley & Sons, Ltd. 相似文献
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Periodic outbreaks of hand, foot and mouth disease(HFMD) occur in children under 5 years old, and can cause death in some cases. The C4 strain of enterovirus 71(EV71) is the main pathogen that causes HFMD in China. Although no drugs against EV71 are available, some studies have shown that candidate vaccines or viral capsid proteins can produce anti-EV71 immunity. In this study, female BABL/c mice(6–8 weeks old) were immunized with virus-like particles(VLPs) of EV71 produced in yeast to screen for anti-EV71 antibodies. Two hybridomas that could produce neutralizing antibodies against EV71 were obtained. Both neutralizing m Abs(D4 and G12) were confirmed to bind the VP1 capsid protein of EV71, and could protect 95% cells from 100 TCID50 EV71 infection at 25 μg/m L solution(lowest concentration). Those two neutralizing m Abs identified in the study may be promising candidates in development for m Abs to treat EV71 infection, and utilized as suitable reagents for use in diagnostic tests and biological studies. 相似文献
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Huajun Li Shuxian Li Jianfeng Zheng Chunyan Cai Bin Ye Jun Yang Zhimin Chen 《Microbiology and immunology》2015,59(3):152-159
Enterovirus 71 (EV71) infection can cause severe neurological complications including meningoencephalitis (ME) in some patients with hand, foot and mouth disease (HFMD). However, to date no studies have reported changes in cytokine concentrations and their correlations with clinical variables in patients with ME following EV71 infection. In this study, responses of Th1/Th2 cytokine, including IL‐2, IL‐4, IL‐6, IL‐10, TNF‐α and IFN‐γ, in cerebrospinal fluid (CSF) from patients with EV71‐related HFMD with ME and patients with febrile convulsions (FC) were analyzed using cytometric bead array technology. It was found that CSF IL‐6 and IFN‐γ concentrations were significantly higher in patients with EV71‐related ME than in those with FC. Additionally, both CSF IL‐6 and IFN‐γ concentrations were correlated with CSF cytology, fever duration and duration of hospital stay. More interestingly, a positive correlation between CSF IL‐6 and IFN‐γ concentrations was observed. Finally, receiver operating characteristic analysis revealed that when a cutoff value of 9.40 pg/mL was set for IL‐6, the sensitivity and specificity were 84.5% and 85.5%, respectively, for discriminating EV71‐related ME from FC. In conclusion, IL‐6 and IFN‐γ may be associated with EV71‐induced neuropathology. 相似文献
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Rong-Rong Zhang Meng-Jiao He Chao Zhou Yan-Peng Xu Wei Tang Tian-Shu Cao Zheng-Jian Wang Mei Wu Tao Ming Yi-Jiao Huang Meng-Xu Sun Hui Zhao Yong-Qiang Deng Xiao-Feng Li Bin Wang Qing Ye Cheng-Feng Qin 《中国病毒学》2024,39(5):812-820
Human Enterovirus 71 (EV71) has emerged as one of the predominant causative agents of hand, foot and mouth disease (HFMD) with global impact. Despite the inactivated vaccine being licensed, other vaccine candidates based on advanced technology platforms are under development. In this report, we rationally designed and constructed two DNA-launched live attenuated vaccine candidates (pDL-EV71) under the control of specific promoters. In vitro and in vivo transfection with pDL-EV71 driven by the CMV promoter successfully yielded fully infectious EV71. More importantly, the administration of pDL-EV71 did not cause clinical symptoms following intracranial or intramuscular inoculation in neonatal and IFNα/βR-/- mice, demonstrating its safety profile. Moreover, a single-dose or two-dose immunization with pDL-EV71 elicited robust neutralizing antibodies against EV71 as well as an antigen-specific cellular response in mice. A single-dose immunization with 10 μg of pDL-EV71 conferred complete protection against lethal EV71 infection in neonates born to immunized maternal mice. Overall, our present results demonstrate that pDL-EV71 is a safe and effective vaccine candidate against EV71 for further development. 相似文献
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Zhihui Li Bin Cui Xiaowen Liu Laicheng Wang Qingjie Xian Zhaoxi Lu Shuntao Liu Yinguang Cao Yueran Zhao 《Microbiology and immunology》2020,64(3):189-201
Enterovirus 71 (EV71) is the predominant pathogen for severe hand, foot, and mouth disease (HFMD) in children younger than 5 years, and currently no effective drugs are available for EV71. Thus, there is an urgent need to develop new drugs for the control of EV71 infection. In this study, LJ04 was extracted from Laminaria japonica using diethylaminoethyl cellulose-52 with 0.4 mol/l NaCl as the eluent, and its virucidal activity was evaluated based on its cytopathic effects on a microplate. LJ04 is composed of fucose, galactose, and mannose and mainly showed good virucidal activity against EV71. The antiviral mechanisms of LJ04 were the direct inactivation of the virus, the blockage of virus binding, disruptions to viral entry, and weak inhibitory activity against the nonstructural protein 3C. The two most important findings from this study were that LJ04 inhibited EV71 proliferation in HM1900 cells, which are a human microglia cell line, and that LJ04 can directly inactivate EV71 within 2 hr at 37°C. This study demonstrates for the first time the ability of a polysaccharide from L. japonica to inhibit viral and 3C activity; importantly, the inhibition of 3C might have a minor effect on the antiviral effect of LJ04. Consequently, our results identify LJ04 as a potential drug candidate for the control of severe EV71 infection in clinical settings. 相似文献
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Enterovirus 71 (EV71) is a major aetiological agent of hand, foot and mouth disease (HFMD). In recent years, several outbreaks in East Asia were associated with neurological complications and numerous deaths. An outbreak in Singapore in October 2000 afflicted thousands of children, resulting in four fatal cases from three of whom EV71 was isolated. The genomes of two representative EV71 strains isolated from a fatal case and a surviving patient were completely sequenced, and their nucleotide and amino acid sequences compared with known EV71 strains. The two outbreak strains were classified under genogroup B, together with those previously isolated in Singapore, Malaysia and Japan. Comparative sequence analysis of the two Singapore strains revealed 99% nucleotide similarity, while their deduced amino acid sequences were almost identical except for residue 1506 in the 3A non-structural region. Given that the outbreak involved closely related genetic variants of EV71, the broad spectrum of disease severity may be attributed to critical factors such as varying viral inoculation doses or differing host immune responses following infection, but is less likely to be due to the emergence of EV71 strains with heightened virulence. 相似文献
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目的了解濮阳市手足口病流行病学特征,为制定防控策略提供科学依据。方法通过国家疾病监测信息管理系统收集的全市2008—2012年6月6日手足口病疫情资料进行描述和分析,并对部分病例和重症病例标本进行肠道病毒病原学检测。结果全市共报手足口病16 492例,发病高峰是每年的3-5月(第12～20周),呈典型的单峰型曲线;发病年龄以0～4岁居多;男性多于女性;散居儿童多于托幼机构儿童,爆发病例多发生在托幼机构,手足口病病原有EV71、CoxA16和其他肠道病毒,以EV71和CoxA16为主。结论手足口病发病有明显的季节性、年龄和性别差异,小年龄组儿童是手足口病预防控制重点人群,流行年度和流行季节的优势毒株为EV71,重症患者中EV71占到86.35%;非流行年和季节手足病例主要由CoxA16和其他肠道病毒引起。手足口病防控重点应体现在对病例分类管理上,同时应继续加强重症病例疫情监测和爆发控制。 相似文献
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为探讨肠道病毒71型(EV71)VP4基因序列与手足口病(HFMD)不同临床类型之间的关系,分析重组蛋白EV71 VP4的抗原性,并初步探讨其与柯萨奇病毒A16(CA16)重组蛋白VP4是否存在交叉反应性,对2007~2009年从北京患HFMD儿童标本分离到的10株EV71的VP4基因进行克隆测序,运用生物学软件对测序结果进行比较分析,并选择其中1株与1株同期分离的CA16的VP4分别进行原核表达;用表达产物对189份正常体检的成人及来首都儿科研究所就医的非HFMD患儿血清中的IgG进行Western Blot检测,并分析14份确诊为EV71感染和12份CA16感染患者急性期血清中的IgM抗体。分析表明这10株EV71 VP4基因核苷酸同源性为94.20%~100.00%,所推导的氨基酸序列则完全相同,从重症与轻症患儿分离的毒株之间VP4的核苷酸序列未见一致性的差异,基于EV71 VP4基因核苷酸序列的进化树分析表明2007~2009年北京地区所流行的毒株均属于C4亚型;本研究中EV71和CA16的VP4核苷酸序列的同源性为69.60%,所推导的氨基酸序列的同源性为78.60%,在运用Western Blot检测189份血清中的VP4特异性IgG时,EV71 VP4的血清阳性率为38.10%,说明其具有良好的抗原性,CA16 VP4的血清阳性率为58.20%,两者差别具有显著统计学意义(2χ=15.30,P<0.01),提示EV71 VP4与CA16 VP4没有交叉反应性;在用表达的VP4检测已确诊为相应病毒的特异性IgM时,两者皆为阴性,提示感染后机体对VP1和VP4产生不同的反应。 相似文献
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Li-chun WANG Song-qing TANG Yan-mei LI Hong-lin ZHAO Cheng-hong DONG Ping-fang CUI Shao-hui MA Yun LIAO Long-ding LIU Qi-han LI 《Virologica Sinica》2010,(2)
The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates s... 相似文献