This study bioinformatically analyzed the non-VP1 capsid proteins（VP2-VP4） of Coxasckievirus A6（CVA6）, with an attempt to predict their basic physicochemical properties, structural/functional features and linear B...This study bioinformatically analyzed the non-VP1 capsid proteins（VP2-VP4） of Coxasckievirus A6（CVA6）, with an attempt to predict their basic physicochemical properties, structural/functional features and linear B cell eiptopes. The online tools Sub Loc, Target P and the others from Ex PASy Bioinformatics Resource Portal, and SWISS-MODEL（an online protein structure modeling server）, were utilized to analyze the amino acid（AA） sequences of VP2-VP4 proteins of CVA6. Our results showed that the VP proteins of CVA6 were all of hydrophilic nature, contained phosphorylation and glycosylation sites and harbored no signal peptide sequences and acetylation sites. Except VP3, the other proteins did not have transmembrane helix structure and nuclear localization signal sequences. Random coils were the major conformation of the secondary structure of the capsid proteins. Analysis of the linear B cell epitopes by employing Bepipred showed that the average antigenic indices（AI） of individual VP proteins were all greater than 0 and the average AI of VP4 was substantially higher than that of VP2 and VP3. The VP proteins all contained a number of potential B cell epitopes and some eiptopes were located at the internal side of the viral capsid or were buried. We successfully predicted the fundamental physicochemical properties, structural/functional features and the linear B cell eiptopes and found that different VP proteins share some common features and each has its unique attributes. These findings will help us understand the pathogenicity of CVA6 and develop related vaccines and immunodiagnostic reagents.展开更多
BACKGROUND Hand,foot,and mouth disease(HFMD)has become one of the most common infectious diseases in China.Before 2016,the primary causal serotypes were enterovirus A71(EV-A71)and coxsackievirus A16(CV-A16).Following ...BACKGROUND Hand,foot,and mouth disease(HFMD)has become one of the most common infectious diseases in China.Before 2016,the primary causal serotypes were enterovirus A71(EV-A71)and coxsackievirus A16(CV-A16).Following the introduction of EV-A71 vaccines in China since 2016,the situation could change.CV-A6 has recently replaced EV-A71 and CV-A16 in some areas of China.However,the epidemiological characteristics of central China remain unknown.AIM To investigate the clinical symptoms and pathogen spectrum of HFMD in Shiyan City,central China,in recent years.METHODS The epidemiological,clinical,and laboratory data from HFMD cases reported to the Shiyan Center for Disease Control and Prevention between January 2016 and December 2020 were analyzed.196 throat swab specimens were collected from hospitalized HFMD patients between January 2018 and December 2020.To detect and genotype enteroviruses,real-time reverse transcription-polymerase chain reaction and sequencing of the 5'-untranslated region were used.In Shiyan,168 laboratory-confirmed HFMD cases were studied using a logistic regression model to determine the effect of predominant enterovirus serotypes.Based on the logistic regression model,the least absolute shrinkage and selection operator model was used to analyze the correlation between CV-A6 infection and various clinical characteristics in HFMD patients in Shiyan.RESULTS From 2016 to 2020,35840 HFMD cases were reported in Shiyan.The number of cases decreased by 48.4%from 2016 to 2017.Approximately 1.58-fold increases were found in 2018 and 2019 when compared to the previous year,respectively.In 2020,a decrease of about 85.5%was reported when compared to 2019.The most common serotypes shifted from EV-A71 and CV-A16(about 60%-80%in 2016 and 2018)to others(more than 80.0%in 2017,2019,and 2020).EV-A71 lost its dominance in 2017 in Shiyan.Among 196 confirmed HFMD cases,85.7%tested positive for enterovirus,with CV-A6 being the most common serotype(121/168,72.0%).The positive rates for CV-A16 and CVA10 were 4.8%and 3.0%,respectively.There was no EV-A71 discovered.Infection with CV-A6 was linked to fever,myocardial damage,increased creatine kinase MB isoenzyme,and lactate dehydrogenase levels.CONCLUSION CV-A6 was the most common enterovirus serotype in Shiyan City,replacing EV-A71 and CV-A16 as the HFMD pathogen.Developing vaccines against CV-A6 or multiple pathogens,as well as rising CV-A6 surveillance,will help prevent HFMD in central China.展开更多
Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infection...Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infections. Coxsackievirus B6 (CV-B6)belongs to the species EV-B, which currently consists of 63 serotypes, including echovirus (serotypes 1–7,9, 11–21, 24–27, 29–33), coxsackievirus group A (CVA9), coxsackievirus group B (CV-B, serotypes 1–6),the newly identified EVs (serotypes EV-B69, B73–75,B77–88, B93, B97–98, B100–101, B106–107, and B110–113), and the simian enterovirus SA5.展开更多
Coxsackievirus A6 of the D3a genotype(CVA6 D3a)is a primary pathogen causingmainland of China's hand,foot,and mouth disease(HFMD).Viral‐like particle(VLP)vaccines represent a potential candidate vaccine to preven...Coxsackievirus A6 of the D3a genotype(CVA6 D3a)is a primary pathogen causingmainland of China's hand,foot,and mouth disease(HFMD).Viral‐like particle(VLP)vaccines represent a potential candidate vaccine to prevent HFMD.This study collected Anti‐CVA6 D3a VLPs serum from BALB/c female mice immunized using CVA6 D3a VLPs.The neutralizing antibody levels were compared against the representative 14‐JX2018(D3a)and N4‐YN2015(D3b)strains between the antisera of different immune pathways.The immunoprotective effect of anti‐CVA6 D3a VLPs against these strains was monitored using pathological sections and immuno-histochemical results of lung and skeletal muscle tissues in seven‐day‐old Institute of Cancer Research(ICR)mice.Immunological protection against different branches of viruses was evaluated in 7‐day‐old(serum passive immune protection)and 14‐day‐old(VLPs active immune protection)neonatal ICR mice models.Serum‐neutralizing antibody levels were positively correlated with the number of immunizations and higher against 14‐JX2018 than against N4‐YN2015.Furthermore,these levels were significantly higher with abdominal injection than intramuscular injection.The immunized serum of 7‐day‐old ICR mice inoculated three times was 100%protected against CVA6 D3a 14‐JX2018(lethal titer:106.25 TCID 50)and CVA6 D3b N4‐YN2015(lethal titer:105.25TCID 50)fatal attacks,respectively.For ICR mice that have completed two active immunizations for 14 days,both CVA6 D3a 14‐JX2015(challenge titer:108.25 TCID 50)and CVA6 D3b N4‐YN2015(challenge titer:107.25 TCID 50)were used for the challenge,and the mice were able to survive.Overall,the CVA6 D3a VLPs prepared in this study are a potential vaccine candidate for CVA6,as it has the optimal protective effect against both CVA6 D3a and D3b evolutionary branches viruses.展开更多
目的本研究旨在探究柯萨奇病毒A组16型(Coxsackievirus A 16,CVA16)感染后是否会影响N6-甲基腺嘌呤(N6-methyladenosine,m6A)甲基化相关蛋白在人呼吸道上皮细胞(16HBE)、ICR乳鼠和SCARB2人源化小鼠中的表达,以及在细胞中的定位。方法病...目的本研究旨在探究柯萨奇病毒A组16型(Coxsackievirus A 16,CVA16)感染后是否会影响N6-甲基腺嘌呤(N6-methyladenosine,m6A)甲基化相关蛋白在人呼吸道上皮细胞(16HBE)、ICR乳鼠和SCARB2人源化小鼠中的表达,以及在细胞中的定位。方法病毒分别以感染复数(MOI)=0.1感染16HBE和107 CCID50/ml感染小鼠,Western blot分析甲基转移酶、去甲基化酶和甲基化阅读蛋白的表达变化,免疫荧光观察这些蛋白质在细胞中的定位。结果研究结果发现,随着CVA16病毒感染时间的增加,m6A甲基化相关蛋白在细胞中表达水平逐渐下调,在ICR乳鼠和SCARB2小鼠中无明显变化;病毒感染后,m6A甲基化相关蛋白在细胞核与细胞质中重新分布,甚至发生降解。结论CVA16在宿主细胞中复制时可以改变m6A甲基化修饰相关蛋白的表达和细胞定位。本研究结果提示m6A修饰可能是肠道病毒治疗的潜在新靶点。展开更多
本研究旨在通过对手足口病(Hand,foot and mouth disease,HFMD)病例的病原学标本进行肠道病毒分型鉴定,和对CVA6和CVA10开展VP1基因全长序列分析,探究广东省汕头市HFMD病原谱及CVA6和CVA10基因进化特征,为当地HFMD肠道病毒监测及预警提...本研究旨在通过对手足口病(Hand,foot and mouth disease,HFMD)病例的病原学标本进行肠道病毒分型鉴定,和对CVA6和CVA10开展VP1基因全长序列分析,探究广东省汕头市HFMD病原谱及CVA6和CVA10基因进化特征,为当地HFMD肠道病毒监测及预警提供一定的技术支撑。将汕头市病原监测哨点医院2018-2021年的HFMD样本进行RT-PCR肠道病毒病原学分型,筛选出CVA6和CVA10样本进行VP1区全长基因扩增和测序,并通过系统进化分析、同源性和突变位点分析了解其遗传进化特征。结果显示,2018-2021年共监测到HFMD阳性样本706份,其主要病原体为CVA6和CVA10。CVA6和CVA10流行株分别属于D3和C2亚型。CVA6样本VP1核苷酸和氨基酸的相似性分别为91.37%~100.00%和92.79%~100.00%,与2010年的中国台湾株高度同源(GenBank:JQ946055.1)。CVA10样本VP1核苷酸和氨基酸的相似性分别为92.28%~100.00%和96.98%~100.00%,与2014年的广东株高度同源(GenBank:MN451243.1)。与同型别原型株相比,CVA6样本有2个高频的非同义氨基酸突变位点造成了氨基酸极性或带电性的改变(F305S、T14A),CVA10样本则有9个(D25N、A33T、F148Y、Q223L、A291S、A78T、N27E、V240E、V107T)。以上结果提示当地需密切关注病毒流行谱和动态监测HFMD患者临床症状,以防病毒蛋白结构和功能的改变,加重其毒性和感染力。展开更多
基金supported by the National Natural Science Foundation of China(No.81460304)Guangxi Natural Science Foundation(No.2015GXNSFDA139020)a research program sponsored by the Health Bureau of Guangxi Zhuang Autonomous Region,China(No.Z2014298)
文摘This study bioinformatically analyzed the non-VP1 capsid proteins（VP2-VP4） of Coxasckievirus A6（CVA6）, with an attempt to predict their basic physicochemical properties, structural/functional features and linear B cell eiptopes. The online tools Sub Loc, Target P and the others from Ex PASy Bioinformatics Resource Portal, and SWISS-MODEL（an online protein structure modeling server）, were utilized to analyze the amino acid（AA） sequences of VP2-VP4 proteins of CVA6. Our results showed that the VP proteins of CVA6 were all of hydrophilic nature, contained phosphorylation and glycosylation sites and harbored no signal peptide sequences and acetylation sites. Except VP3, the other proteins did not have transmembrane helix structure and nuclear localization signal sequences. Random coils were the major conformation of the secondary structure of the capsid proteins. Analysis of the linear B cell epitopes by employing Bepipred showed that the average antigenic indices（AI） of individual VP proteins were all greater than 0 and the average AI of VP4 was substantially higher than that of VP2 and VP3. The VP proteins all contained a number of potential B cell epitopes and some eiptopes were located at the internal side of the viral capsid or were buried. We successfully predicted the fundamental physicochemical properties, structural/functional features and the linear B cell eiptopes and found that different VP proteins share some common features and each has its unique attributes. These findings will help us understand the pathogenicity of CVA6 and develop related vaccines and immunodiagnostic reagents.
基金Supported by the Hubei Province Health and Family Planning A Scientific Research Project,No.WJ2017M220the Wuhan Health Bureau Scientific Research Fund,No.WX19C11+2 种基金the Joint Precision Medical Research Fund From Taihe Hospital,No.2016JZ10the Shiyan COVID-19 Pilot Emergency Scientific Research Project,No.20Y19the Wuhan Children's Hospital Research Project,No.2017FE007.
文摘BACKGROUND Hand,foot,and mouth disease(HFMD)has become one of the most common infectious diseases in China.Before 2016,the primary causal serotypes were enterovirus A71(EV-A71)and coxsackievirus A16(CV-A16).Following the introduction of EV-A71 vaccines in China since 2016,the situation could change.CV-A6 has recently replaced EV-A71 and CV-A16 in some areas of China.However,the epidemiological characteristics of central China remain unknown.AIM To investigate the clinical symptoms and pathogen spectrum of HFMD in Shiyan City,central China,in recent years.METHODS The epidemiological,clinical,and laboratory data from HFMD cases reported to the Shiyan Center for Disease Control and Prevention between January 2016 and December 2020 were analyzed.196 throat swab specimens were collected from hospitalized HFMD patients between January 2018 and December 2020.To detect and genotype enteroviruses,real-time reverse transcription-polymerase chain reaction and sequencing of the 5'-untranslated region were used.In Shiyan,168 laboratory-confirmed HFMD cases were studied using a logistic regression model to determine the effect of predominant enterovirus serotypes.Based on the logistic regression model,the least absolute shrinkage and selection operator model was used to analyze the correlation between CV-A6 infection and various clinical characteristics in HFMD patients in Shiyan.RESULTS From 2016 to 2020,35840 HFMD cases were reported in Shiyan.The number of cases decreased by 48.4%from 2016 to 2017.Approximately 1.58-fold increases were found in 2018 and 2019 when compared to the previous year,respectively.In 2020,a decrease of about 85.5%was reported when compared to 2019.The most common serotypes shifted from EV-A71 and CV-A16(about 60%-80%in 2016 and 2018)to others(more than 80.0%in 2017,2019,and 2020).EV-A71 lost its dominance in 2017 in Shiyan.Among 196 confirmed HFMD cases,85.7%tested positive for enterovirus,with CV-A6 being the most common serotype(121/168,72.0%).The positive rates for CV-A16 and CVA10 were 4.8%and 3.0%,respectively.There was no EV-A71 discovered.Infection with CV-A6 was linked to fever,myocardial damage,increased creatine kinase MB isoenzyme,and lactate dehydrogenase levels.CONCLUSION CV-A6 was the most common enterovirus serotype in Shiyan City,replacing EV-A71 and CV-A16 as the HFMD pathogen.Developing vaccines against CV-A6 or multiple pathogens,as well as rising CV-A6 surveillance,will help prevent HFMD in central China.
基金supported by the National Key Science and Technology Projects of China [Project Nos.2017ZX10104001,2018ZX10711001,and 2018ZX10713002]
文摘Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infections. Coxsackievirus B6 (CV-B6)belongs to the species EV-B, which currently consists of 63 serotypes, including echovirus (serotypes 1–7,9, 11–21, 24–27, 29–33), coxsackievirus group A (CVA9), coxsackievirus group B (CV-B, serotypes 1–6),the newly identified EVs (serotypes EV-B69, B73–75,B77–88, B93, B97–98, B100–101, B106–107, and B110–113), and the simian enterovirus SA5.
基金supported by the National Key Researchand Development Programof China (Project No.2021YFC2302003).
文摘Coxsackievirus A6 of the D3a genotype(CVA6 D3a)is a primary pathogen causingmainland of China's hand,foot,and mouth disease(HFMD).Viral‐like particle(VLP)vaccines represent a potential candidate vaccine to prevent HFMD.This study collected Anti‐CVA6 D3a VLPs serum from BALB/c female mice immunized using CVA6 D3a VLPs.The neutralizing antibody levels were compared against the representative 14‐JX2018(D3a)and N4‐YN2015(D3b)strains between the antisera of different immune pathways.The immunoprotective effect of anti‐CVA6 D3a VLPs against these strains was monitored using pathological sections and immuno-histochemical results of lung and skeletal muscle tissues in seven‐day‐old Institute of Cancer Research(ICR)mice.Immunological protection against different branches of viruses was evaluated in 7‐day‐old(serum passive immune protection)and 14‐day‐old(VLPs active immune protection)neonatal ICR mice models.Serum‐neutralizing antibody levels were positively correlated with the number of immunizations and higher against 14‐JX2018 than against N4‐YN2015.Furthermore,these levels were significantly higher with abdominal injection than intramuscular injection.The immunized serum of 7‐day‐old ICR mice inoculated three times was 100%protected against CVA6 D3a 14‐JX2018(lethal titer:106.25 TCID 50)and CVA6 D3b N4‐YN2015(lethal titer:105.25TCID 50)fatal attacks,respectively.For ICR mice that have completed two active immunizations for 14 days,both CVA6 D3a 14‐JX2015(challenge titer:108.25 TCID 50)and CVA6 D3b N4‐YN2015(challenge titer:107.25 TCID 50)were used for the challenge,and the mice were able to survive.Overall,the CVA6 D3a VLPs prepared in this study are a potential vaccine candidate for CVA6,as it has the optimal protective effect against both CVA6 D3a and D3b evolutionary branches viruses.
文摘目的本研究旨在探究柯萨奇病毒A组16型(Coxsackievirus A 16,CVA16)感染后是否会影响N6-甲基腺嘌呤(N6-methyladenosine,m6A)甲基化相关蛋白在人呼吸道上皮细胞(16HBE)、ICR乳鼠和SCARB2人源化小鼠中的表达,以及在细胞中的定位。方法病毒分别以感染复数(MOI)=0.1感染16HBE和107 CCID50/ml感染小鼠,Western blot分析甲基转移酶、去甲基化酶和甲基化阅读蛋白的表达变化,免疫荧光观察这些蛋白质在细胞中的定位。结果研究结果发现,随着CVA16病毒感染时间的增加,m6A甲基化相关蛋白在细胞中表达水平逐渐下调,在ICR乳鼠和SCARB2小鼠中无明显变化;病毒感染后,m6A甲基化相关蛋白在细胞核与细胞质中重新分布,甚至发生降解。结论CVA16在宿主细胞中复制时可以改变m6A甲基化修饰相关蛋白的表达和细胞定位。本研究结果提示m6A修饰可能是肠道病毒治疗的潜在新靶点。
文摘本研究旨在通过对手足口病(Hand,foot and mouth disease,HFMD)病例的病原学标本进行肠道病毒分型鉴定,和对CVA6和CVA10开展VP1基因全长序列分析,探究广东省汕头市HFMD病原谱及CVA6和CVA10基因进化特征,为当地HFMD肠道病毒监测及预警提供一定的技术支撑。将汕头市病原监测哨点医院2018-2021年的HFMD样本进行RT-PCR肠道病毒病原学分型,筛选出CVA6和CVA10样本进行VP1区全长基因扩增和测序,并通过系统进化分析、同源性和突变位点分析了解其遗传进化特征。结果显示,2018-2021年共监测到HFMD阳性样本706份,其主要病原体为CVA6和CVA10。CVA6和CVA10流行株分别属于D3和C2亚型。CVA6样本VP1核苷酸和氨基酸的相似性分别为91.37%~100.00%和92.79%~100.00%,与2010年的中国台湾株高度同源(GenBank:JQ946055.1)。CVA10样本VP1核苷酸和氨基酸的相似性分别为92.28%~100.00%和96.98%~100.00%,与2014年的广东株高度同源(GenBank:MN451243.1)。与同型别原型株相比,CVA6样本有2个高频的非同义氨基酸突变位点造成了氨基酸极性或带电性的改变(F305S、T14A),CVA10样本则有9个(D25N、A33T、F148Y、Q223L、A291S、A78T、N27E、V240E、V107T)。以上结果提示当地需密切关注病毒流行谱和动态监测HFMD患者临床症状,以防病毒蛋白结构和功能的改变,加重其毒性和感染力。
