Molecular Epidemiology of Coxsackievirus B1-5 Associated with HFMD in Fujian Province, China, 2011-2016

Hand,foot and mouth disease(HFMD)is a common infectious disease that usually affects children less than 5 years of age.HFMD is caused by human enteroviruses(HEVs).HEVs,members of the Enterovirus genus of the Picornaviridae(small RNA virus)family.

辽宁省首株柯萨奇病毒B组5型全基因组序列分析

研究引起辽宁地区手足口病的柯萨奇病毒B组5型(coxsackievirus B5,CV-B5)基因组特征。对2018年从辽宁省688份肠道病毒核酸阳性的标本中分离到的1株CV-B5进行高通量测序,并对其全基因组进行遗传进化分析。结果表明,CV-B5辽宁分离株与国内流行株的全基因组核苷酸序列同源性为78.5%~97%,氨基酸序列同源性为75.3%~96.7%。基于全基因组的进化分析将CV-B5流行株分为A~D四个基因型,辽宁分离株属于D基因型。通过重组分析发现其在P3区的3D区段发生重组。首次在辽宁地区手足口病患儿中分离出CV-B5,辽宁省分离株(LN2018-23-21/CHN/2018)可能为重组株。

Exosomal let-7a-5p derived from human umbilical cord mesenchymal stem cells alleviates coxsackievirus B3-induced cardiomyocyte ferroptosis via the SMAD2/ZFP36 signal axis

Viral myocarditis(VMC)is one of the most common acquired heart diseases in children and teenagers.However,its pathogenesis is still unclear,and effective treatments are lacking.This study aimed to investigate the regulatory pathway by which exosomes alleviate ferroptosis in cardiomyocytes(CMCs)induced by coxsackievirus B3(CVB3).CVB3 was utilized for inducing the VMC mouse model and cellular model.Cardiac echocardiography,left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS)were implemented to assess the cardiac function.In CVB3-induced VMC mice,cardiac insufficiency was observed,as well as the altered levels of ferroptosis-related indicators(glutathione) peroxidase 4(GPX4),glutathione(GSH),and malondialdehyde(MDA).However,exosomes derived from human umbilical cord mesenchymal stem cells(hucMSCs-exo)could restore the changes caused by CVB3 stimulation.Let-7a-5p was enriched in hucMSCs-exo,and the inhibitory ffect of hucMSCs-exoa-ie-pmimo on CVB3-induced ferroptosis was higher than that of hucMSCs-exommie N(NC:negative control).Mothers against decapentaplegic homolog 2(SMAD2)increased in the VMC group,while the expression of zinc-finger protein 36(ZFP36)decreased.Let-7a-5p was confirmed to interact with SMAD2 messenger RNA(mRNA),and the SMAD2 protein interacted directly with the ZFP36 protein.Silencing SMAD2 and overexpressing ZFP36 inhibited the expression of ferroptosis-related indicators.Meanwhile,the levels of GPX4,solute carrier family 7,member 11(SLC7A11),and GSH were lower in the SMAD2 overexpression plasmid(oe-SMAD2)+let-7a-5p mimic group than in the oe-NC+let-7a-5p mimic group,while those of MDA,reactive oxygen species(ROS),and Fe^(2+)increased.In conclusion,these data showed that ferroptosis could be regulated by mediating SMAD2 expression.Exo-let-7a-5p derived from hucMSCs could mediate SMAD2 to promote the expression of ZFP36,which further inhibited the ferroptosis of CMCs to alleviate CVB3-induced VMC.

Long noncoding RNA 1392 regulates MDA5 by interaction with ELAVL1 to inhibit coxsackievirus B5 infection

Long noncoding RNAs(lncRNAs)modulate many aspects of biological and pathological processes.Recent studies have shown that host lncRNAs participate in the antiviral immune response,but functional lncRNAs in coxsackievirus B5(CVB5)infection remain unknown.Here,we identified a novel cytoplasmic lncRNA,LINC1392,which was highly inducible in CVB5 infected RD cells in a time-and dose-dependent manner,and also can be induced by the viral RNA and IFN-β.Further investigation showed that LINC1392 promoted several important interferon-stimulated genes(ISGs)expression,including IFIT1,IFIT2,and IFITM3 by activating MDA5,thereby inhibiting the replication of CVB5 in vitro.Mechanistically,LINC1392 bound to ELAV like RNA binding protein 1(ELAVL1)and blocked ELAVL1 interaction with MDA5.Functional study revealed that the 245–835 nt locus of LINC1392 exerted the antiviral effect and was also an important site for ELAVL1 binding.In mice,LINC1392 could inhibit CVB5 replication and alleviated the histopathological lesions of intestinal and brain tissues induced by viral infection.Our findings collectively reveal that the novel LINC1392 acts as a positive regulator in the IFN-I signaling pathway against CVB5 infection.Elucidating the underlying mechanisms on how lncRNA regulats the host innate immunity response towards CVB5 infection will lay the foundation for antiviral drug research.

一株引起病毒性脑炎的柯萨奇B组5型病毒全基因组分析

目的对一株病毒性脑炎病原柯萨奇病毒B5(coxsackievirus B5,CV-B5)分离株KMA2/YN/CHN/2010的全基因组及其遗传特性进行分析。方法对297份病毒性脑炎患者脑脊液在RD细胞进行病毒分离;提取病毒RNA,采用RT-PCR移步法分段扩增CV-B5全长基因,经测序和拼接后,利用MEGA 7.0和SimPlot 3.5.1等软件进行分析。结果通过分离获得一株阳性分离株,经鉴定为CV-B5 KMA2/YN/CHN/2010株,其基因组全长为7 395 bp,编码含2 185个氨基酸残基的多聚蛋白。与其他CV-B5病毒分离株的全基因组的核苷酸和氨基酸同源性分别为79.6%~98.8%和93.1%~98.1%;KMA2/YN/CHN/2010与其他CV-B5株在各区段核苷酸和氨基酸同源性分别为75.0%~100.0%和88.8%~100.0%。其中与A210/KM/09株同源性最高。进化分析发现CV-B5分4个基因型,KMA2/YN/CHN/2010属于C基因型。通过重组分析发现KMA2/YN/CHN/2010可能为重组株。结论 KMA2/YN/CHN/2010与其他中国株一样同属于C基因型。

2014年山东省柯萨奇病毒B5型的分子流行病学研究

为了解山东省柯萨奇病毒B5型(Coxsackievirus B5,CV-B5)的分子进化特征,本研究对山东省2014年1~12月无菌性脑膜炎(Aseptic meningitis,AM)病例标本和环境污水监测标本进行肠道病毒(Enterovirus,EV)分离,阳性分离物采用分子生物学方法鉴定型别,并对CV-B5的VP1区序列进行系统进化树分析。结果显示,2014年AM病例中分离到69株CV-B5,占EV分离株43.95%;环境污水中分离到159株CV-B5,占非脊髓灰质炎EV分离株34.49%。上述CV-B5分离株具有季节性特征,夏秋季高峰明显。系统进化树分析表明,本研究分离株属于A、B两个基因簇,各簇之内AM和环境污水分离株之间有密切的亲缘关系,且环境污水分离株具有一定的地域聚集性。A、B基因簇的组内遗传距离分别为0.026、0.030,A基因簇与B基因簇组间遗传距离为0.085,提示CV-B5在山东地区存在2个传播链共循环,同时可能存在CV-B5跨地区传播和流行。本研究结果证明环境监测具有对EV相关疾病的预警能力。

天津市致病毒性脑炎柯萨奇病毒B组5型全基因组序列分析

肠道病毒是我国病毒性脑炎(Viral encephalitis,VE)的主要病原体。本文研究对4株引起VE的天津柯萨奇病毒B组5型(Coxsackievirus B5,CV-B5)分离株进行Illumina MiniSeq高通量测序,并对其全基因组特征、进化及重组特点进行分析。结果提示,4株CV-B5天津分离株的全基因组核苷酸和氨基酸序列同源性分别为84.5%~100.0%和98.1%~100.0%,与国内流行株的全基因组核苷酸序列同源性为83.2%~96.5%,氨基酸序列同源性为96.4%~99.4%。基于全基因组的系统进化分析将CV-B5流行株分为A-D四个基因型,其中天津与国内流行株均属于C基因型。C基因型进一步分为3个进化分支,而天津分离株处在两个不同的分支上。基于基因组各区段序列的系统进化与SimPlot重组分析结果显示,天津分离株15-39N、15-41N与埃可病毒30型(Echovirus 30,E-30)原型株在P3区3B、3C、3D区域均检测到重组信号。本研究有助于了解CV-B5的全基因组特点和重组规律,为相关疾病的防控提供依据。

Expression Profile and Function Analysis of Long Non-coding RNAs in the Infection of Coxsackievirus B3

The roles of IncRNAs in the infection of enteroviruses have been barely demonstrated.In this study,we used coxsackievirus B3(CVB3),a typical enterovirus,as a model to investigate the expression profiles and functional roles of IncRNAs in enterovirus infection.We profiled IncRNAs and mRNA expression in CVB3-infected HeLa cells by IncRNA-mRNA integrated microarrays.As a result,700 differentially expressed IncRNAs(431 up-regulated and 269 down-regulated)and 665 differentially expressed mRNAs(299 up-regulated and 366 down-regulated)were identified in CVB3 infection.Then we performed IncRNA-mRNA integrated pathway analysis to identify potential functional impacts of the differentially expressed mRNAs,in which IncRNA-mRNA correlation network was built.According to IncRNA-mRNA correlation,we found that XLOC-001188,an IncRNA down-regulated in CVB3 infection,was negatively correlated with NFAT5 mRNA,an anti-CVB3 gene reported previously.This interaction was supported by qPCR detection following siRNA-mediated knockdown of XLOC-001188,which showed an increase of NFAT5 mRNA and a reduction of CVB3 genomic RNA.In addition,we observed that four most significantly altered IncRNAs,SNHG11,RP11-145F16.2,RP11-1023L17.1 and RP11-1021N1.2 share several common correlated genes critical for CVB3 infection,such as BRE and IRF2BP1.In all,our studies reveal the alteration of IncRNA expression in CVB3 infection and its potential influence on CVB3 replication,providing useful information for future studies of enterovirus infection.

2011年龙岩市手足口病例肠道病毒通用引物核酸阳性未分型样本病原分析

目的了解龙岩市大量判定为其他肠道病毒(EV)感染的手足口病(HFMD)样本的病原组成。方法对龙岩市2011年经Real-time RT-PCR检测判为未分型EV感染的104份HFMD咽拭子样本经RD细胞分离培养后以RT-PCR法检测EV、EV71、CoxA16,对仍不能分型的样本以187/222引物进行扩增,对扩增产物进行纯化回收、克隆、测序,并用Blast程序在线比对判定型别。同时对培养物进行Real-time RT-PCR检测EV、EV71、CoxA16。任一法检出培养物阳性即判为阳性。结果 104份样本中检出EV阳性88份(84.62%),包括29份EV71,4份EV71+CoxA16,3份CoxB5,2份CoxA10,50份仍未能分型。结论 Real-time RT-PCR检测未能分型的HFMD病例中,相当部分(37.18%)还是由EV71所致,临床不应该随意排除该类病例EV71感染的可能,同时还包含CoxB5和CoxA10病毒。