A型流感病毒(influenza A virus,IAV)极易发生突变和重组,导致现有疫苗和抗流感药物失效。因此,开发有效的广谱抗流感病毒疫苗和药物是我国乃至全球的重要需求。2025年1月9日,中国农业科学院兰州兽医研究所/兰州大学动物医学与生物安全...A型流感病毒(influenza A virus,IAV)极易发生突变和重组,导致现有疫苗和抗流感药物失效。因此,开发有效的广谱抗流感病毒疫苗和药物是我国乃至全球的重要需求。2025年1月9日,中国农业科学院兰州兽医研究所/兰州大学动物医学与生物安全学院朱启运研究员团队在Nature Communications杂志发表了题为“Influenza A Virus H7 nanobody recognizes a conserved immunodominant epitope on hemagglutinin head and confers heterosubtypic protection”的研究论文。该研究分离并纯化了一株特异性识别A型流感病毒血凝素(HA)的纳米抗体E10,该抗体对不同亚型流感病毒具有跨组中和和体内保护效应。研究鉴定E10靶向HA头部结构域的保守氨基酸位点(K166/S167),并且含有E10表位的多肽分子能够诱导交叉反应性抗体,能够对小鼠提供部分交叉免疫保护。该研究不仅为流感防治提供了新的手段,也为广谱流感疫苗研发提供了新的靶点和希望。展开更多
Influenza A virus(IAV)remains a global public health concern,causing influenza-like illness and severe respiratory tract infections.Two major subtypes,A/pdm09 H1N1 and A/H3N2,circulate globally,and their epidemics are...Influenza A virus(IAV)remains a global public health concern,causing influenza-like illness and severe respiratory tract infections.Two major subtypes,A/pdm09 H1N1 and A/H3N2,circulate globally,and their epidemics are influenced by multiple factors,especially during the COVID-19 pandemic.Based on data from the National Influenza Surveillance Program in China,we analyzed the epidemiological and genomic data in Tianjin collected from 2017 to 2025.A total of 77,473 throat swabs were collected,of which 9144 were IAV-positive.The A/pdm09 H1N1 and A/H3N2 lineages exhibited distinct epidemics across different influenza seasons,with a decline in cases observed during the COVID-19 pandemic.We sequenced the genomes of 128 A/pdm09 H1N1 and 113 A/H3N2 clinical isolates and characterized their temporal evolution and genetic diversity using time-scaled phylogenetic analysis.Additionally,we conducted a genetic risk evaluation of the hemagglutinin and neuraminidase segments,identifying key amino acid residues associated with viral adaptation,transmissibility,virulence,and drug resistance.Moreover,no antigenic variants were found in clinical isolates during the recent influenza seasons,though reduced sensitivity to oseltamivir and zanamivir was observed in individual strains.Our surveillance highlights the epidemiology and evolution of IAV before and after the COVID-19 pandemic in Tianjin.展开更多
Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by dir...Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by directly binding to viral RNA.Here,we examined the role of the host RNA-binding protein nucleophosmin-1(NPM1)in IAV replication.We found that,as a nucleolar phosphoprotein,NPM1 directly binds to viral RNA(vRNA)and inhibits the replication of various subtypes of IAV.NPM1 binding to vRNA competitively reduces the assembly of the viral ribonucleoprotein complex and the viral polymerase activity,thereby reducing the generation of progeny viral RNA and virions.The RNA-binding activity of NPM1,with the key residues T199,T219,T234,and T237,is essential for its anti-influenza function.Taken together,our findings demonstrate that NPM1 acts as an RNA-binding protein and interacts with IAV vRNA to suppress viral replication.展开更多
Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an effic...Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.展开更多
Artemisia annua L.is a medicinal herb with multiple therapeutic applications,whereas its antiinfluenza A virus(IAV)efficiency and mechanism of action are still unclear.Here,we investigated the inhibition activity and ...Artemisia annua L.is a medicinal herb with multiple therapeutic applications,whereas its antiinfluenza A virus(IAV)efficiency and mechanism of action are still unclear.Here,we investigated the inhibition activity and mechanism of A.annua leaf methanol extracts(AALME)against IAV in vitro and in vivo.Our results revealed that AALME exhibits potent anti-IAV activity by interacting with IAV particles.Mechanistically,AALME directly targets the IAV nucleoprotein(NP)protein and abolishes the nuclear import of IAV NP.AALME profoundly suppresses IAV-induced mitochondrial apoptosis via suppressing ROS-mediated AIF-dependent pathways.More importantly,we found that AALME plays a crucial role in protecting mice from IAV infection and mitigating IAV pathogenicity.This current work provides mechanistic insight into the mechanism by which AALME controls IAV infection in vitro and in vivo,potentially contributing to the development of antiviral treatments for IAV infection.展开更多
Avian H9N2 viruses have wide host range among the influenza A viruses.However,knowledge of H9N2 mammalian adaptation is limited.To explore the molecular basis of the adaptation to mammals,we performed serial lung pass...Avian H9N2 viruses have wide host range among the influenza A viruses.However,knowledge of H9N2 mammalian adaptation is limited.To explore the molecular basis of the adaptation to mammals,we performed serial lung passaging of the H9N2 strain A/chicken/Hunan/8.27 YYGK3W3-OC/2018(3W3)in mice and identified six mutations in the hemagglutinin(HA)and polymerase acidic(PA)proteins.Mutations L226Q,T511I,and A528V of HA were responsible for enhanced pathogenicity and viral replication in mice;notably,HA-L226Q was the key determinant.Mutations T97I,I545V,and S594G of PA contributed to enhanced polymerase activity in mammalian cells and increased viral replication levels in vitro and in vivo.PA-T97I increased viral polymerase activity by accelerating the viral polymerase complex assembly.Our findings revealed that the viral replication was affected by the presence of PA-97I and/or PA-545V in combination with a triple-point HA mutation.Furthermore,the double-and triple-point PA mutations demonstrated antagonistic effect on viral replication when combined with HA-226Q.Notably,any combination of PA mutations,along with double-point HA mutations,resulted in antagonistic effect on viral replication.We also observed antagonism in viral replication between PA-545V and PA-97I,as well as between HA-528V and PA-545V.Our findings demonstrated that several antagonistic mutations in HA and PA proteins affect viral replication,which may contribute to the H9N2 virus adaptation to mice and mammalian cells.These findings can potentially contribute to the monitoring of H9N2 field strains for assessing their potential risk in mammals.展开更多
Pulmonary macrophages,such as tissue-resident alveolar and interstitial macrophages and recruited monocyte-derived macrophages,are the major macrophages present in the lungs during homeostasis and diseased conditions....Pulmonary macrophages,such as tissue-resident alveolar and interstitial macrophages and recruited monocyte-derived macrophages,are the major macrophages present in the lungs during homeostasis and diseased conditions.While tissue-resident macrophages act as sentinels of the alveolar space and play an important role in maintaining homeostasis and immune regulation,recruited macrophages accumulate in the respiratory tract after acute viral infections.Despite sharing similar anatomical niches,these macrophages are distinct in terms of their origins,surface marker expression,and transcriptional profiles,which impart macrophages with distinguished characteristics in physi-ological and pathophysiological conditions.In this review,we summarize the current view on these macrophage populations,their shared functions,and what makes them distinct from each other in the context of homeostasis andrespiratoryviral infections.展开更多
流行性感冒(简称“流感”)是由流感病毒引起的急性呼吸道传染疾病,据世界卫生组织统计,流感每年可导致300万~500万严重病例,其中29万~65万病例死亡,给社会带来沉重的经济负担,是一个世界性的公共卫生难题。研究发现宿主细胞中存在多条...流行性感冒(简称“流感”)是由流感病毒引起的急性呼吸道传染疾病,据世界卫生组织统计,流感每年可导致300万~500万严重病例,其中29万~65万病例死亡,给社会带来沉重的经济负担,是一个世界性的公共卫生难题。研究发现宿主细胞中存在多条信号通路参与对流感病毒感染的应答,越来越多的研究表明宿主miRNAs通过直接或间接的方式,在流感病毒感染、复制的不同阶段发挥着重要调控作用。本文综合分析了目前关于宿主细胞miRNA对流感病毒复制调控的研究进展,对不同的miRNA具体的调控机制进行系统地归类总结后发现:甲型流感病毒(Influenza A virus,IAV)的PB1、PB2、NA、NP、M1基因是宿主miRNA直接抑制病毒复制的主要靶基因,而在间接调控过程中宿主miRNA主要作用在RIG-I样受体信号通路,Jak-STAT信号通路和Toll样受体信号通路三条流感病毒应答信号途径中,以上发现将更有助于全面理解宿主miRNA对于流感病毒调控网络和宿主细胞与流感病毒的互作机制。展开更多
神经介素B(neuromedin B,NMB)及受体(NMB receptor,NMBR)通过NF-κB信号通路参与抑制甲型流感病毒(influenza A virus,IAV)H1N1亚型(IAV/H1N1)的感染。但关于NMB和NMBR对NF-κB信号通路相关泛素蛋白酶的调控如何,尚未见报道。为探究NMB...神经介素B(neuromedin B,NMB)及受体(NMB receptor,NMBR)通过NF-κB信号通路参与抑制甲型流感病毒(influenza A virus,IAV)H1N1亚型(IAV/H1N1)的感染。但关于NMB和NMBR对NF-κB信号通路相关泛素蛋白酶的调控如何,尚未见报道。为探究NMB和NMBR调控IAV诱导的NF-κB信号通路相关的泛素蛋白酶表达的影响,本研究基于IAV/H9N2感染sh-NMBR细胞与NMB刺激的A549细胞,采用RT-PCR、qRT-PCR及Western blot(WB)分析NMB和NMBR对H9N2感染引起的NF-κB信号通路相关的E3泛素连接酶Mind bomb-2(MIB2)和Ring Finger Protein 8(RNF8)及去泛素蛋白酶Cylindromatosis(CYLD)的表达变化。结果显示:H9N2感染sh-NMBR细胞中,RNF8和CYLD表达增加,MIB2表达和P65磷酸化水平降低;NMB激活A549细胞中NMBR的表达后,诱导细胞中RNF8和CYLD的表达水平下降,MIB2表达和P65磷酸化水平增加。结果表明:NMB和NMBR通过调控IAV/H9N2感染诱导的泛素蛋白酶的表达和P65活性,从而影响IAV/H9N2感染激活的NF-κB信号通路的功能,该结果为深入研究NMB和NMBR抑制甲型流感病毒感染的作用机制提供了理论基础。展开更多
文摘A型流感病毒(influenza A virus,IAV)极易发生突变和重组,导致现有疫苗和抗流感药物失效。因此,开发有效的广谱抗流感病毒疫苗和药物是我国乃至全球的重要需求。2025年1月9日,中国农业科学院兰州兽医研究所/兰州大学动物医学与生物安全学院朱启运研究员团队在Nature Communications杂志发表了题为“Influenza A Virus H7 nanobody recognizes a conserved immunodominant epitope on hemagglutinin head and confers heterosubtypic protection”的研究论文。该研究分离并纯化了一株特异性识别A型流感病毒血凝素(HA)的纳米抗体E10,该抗体对不同亚型流感病毒具有跨组中和和体内保护效应。研究鉴定E10靶向HA头部结构域的保守氨基酸位点(K166/S167),并且含有E10表位的多肽分子能够诱导交叉反应性抗体,能够对小鼠提供部分交叉免疫保护。该研究不仅为流感防治提供了新的手段,也为广谱流感疫苗研发提供了新的靶点和希望。
基金supported by Tianjin Municipal Health Commission(TJWJ2022ZD010)Tianjin Key Medical Discipline(Specialty)Construction Project.
文摘Influenza A virus(IAV)remains a global public health concern,causing influenza-like illness and severe respiratory tract infections.Two major subtypes,A/pdm09 H1N1 and A/H3N2,circulate globally,and their epidemics are influenced by multiple factors,especially during the COVID-19 pandemic.Based on data from the National Influenza Surveillance Program in China,we analyzed the epidemiological and genomic data in Tianjin collected from 2017 to 2025.A total of 77,473 throat swabs were collected,of which 9144 were IAV-positive.The A/pdm09 H1N1 and A/H3N2 lineages exhibited distinct epidemics across different influenza seasons,with a decline in cases observed during the COVID-19 pandemic.We sequenced the genomes of 128 A/pdm09 H1N1 and 113 A/H3N2 clinical isolates and characterized their temporal evolution and genetic diversity using time-scaled phylogenetic analysis.Additionally,we conducted a genetic risk evaluation of the hemagglutinin and neuraminidase segments,identifying key amino acid residues associated with viral adaptation,transmissibility,virulence,and drug resistance.Moreover,no antigenic variants were found in clinical isolates during the recent influenza seasons,though reduced sensitivity to oseltamivir and zanamivir was observed in individual strains.Our surveillance highlights the epidemiology and evolution of IAV before and after the COVID-19 pandemic in Tianjin.
基金supported by funding from the National Natural Science Foundation of China(U23A20243 and 32272972 to QZ,32172820 to SX)the Major Science and Technology Project of Gansu Province(22ZD6NA001 to SX)+1 种基金the Youth Innovation Program(Y2023QC30)the Agricultural Science and Technology Innovation Program(CAAS-ASTIP-JBGS-20210102 to SX)of the Chinese Academy of Agricultural Sciences.
文摘Influenza A viruses(IAVs)are single-stranded negative-sense RNA viruses that continually challenge animal and human health.In IAV-infected cells,host RNA-binding proteins play key roles in the life cycle of IAV by directly binding to viral RNA.Here,we examined the role of the host RNA-binding protein nucleophosmin-1(NPM1)in IAV replication.We found that,as a nucleolar phosphoprotein,NPM1 directly binds to viral RNA(vRNA)and inhibits the replication of various subtypes of IAV.NPM1 binding to vRNA competitively reduces the assembly of the viral ribonucleoprotein complex and the viral polymerase activity,thereby reducing the generation of progeny viral RNA and virions.The RNA-binding activity of NPM1,with the key residues T199,T219,T234,and T237,is essential for its anti-influenza function.Taken together,our findings demonstrate that NPM1 acts as an RNA-binding protein and interacts with IAV vRNA to suppress viral replication.
基金the Animal Ethics Committee of the Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences(SYXK-2020-0010).
文摘Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.
基金supported by grants from the National Natural Science Foundation of China(32170937)Shenzhen Medical Research Fund(SMRF,A2303015)+3 种基金Pearl River Talent Project of Guangdong Province(2021QN02Y426)Shenzhen Peacock Plan Project(827/000655)to Liang Yethe Yunnan Provincial Key Areas Science and Technology Plan Project(202303AC100025)Yunnan Scholar of Yunling(YNWRYLXZ-2019-019)to Rongping Zhang.
文摘Artemisia annua L.is a medicinal herb with multiple therapeutic applications,whereas its antiinfluenza A virus(IAV)efficiency and mechanism of action are still unclear.Here,we investigated the inhibition activity and mechanism of A.annua leaf methanol extracts(AALME)against IAV in vitro and in vivo.Our results revealed that AALME exhibits potent anti-IAV activity by interacting with IAV particles.Mechanistically,AALME directly targets the IAV nucleoprotein(NP)protein and abolishes the nuclear import of IAV NP.AALME profoundly suppresses IAV-induced mitochondrial apoptosis via suppressing ROS-mediated AIF-dependent pathways.More importantly,we found that AALME plays a crucial role in protecting mice from IAV infection and mitigating IAV pathogenicity.This current work provides mechanistic insight into the mechanism by which AALME controls IAV infection in vitro and in vivo,potentially contributing to the development of antiviral treatments for IAV infection.
基金supported by the National Key Research and Development Program of China(NKPs)(2022YFC2604101)the National Science and Technology Major Project of China(2020ZX10001016-002)。
文摘Avian H9N2 viruses have wide host range among the influenza A viruses.However,knowledge of H9N2 mammalian adaptation is limited.To explore the molecular basis of the adaptation to mammals,we performed serial lung passaging of the H9N2 strain A/chicken/Hunan/8.27 YYGK3W3-OC/2018(3W3)in mice and identified six mutations in the hemagglutinin(HA)and polymerase acidic(PA)proteins.Mutations L226Q,T511I,and A528V of HA were responsible for enhanced pathogenicity and viral replication in mice;notably,HA-L226Q was the key determinant.Mutations T97I,I545V,and S594G of PA contributed to enhanced polymerase activity in mammalian cells and increased viral replication levels in vitro and in vivo.PA-T97I increased viral polymerase activity by accelerating the viral polymerase complex assembly.Our findings revealed that the viral replication was affected by the presence of PA-97I and/or PA-545V in combination with a triple-point HA mutation.Furthermore,the double-and triple-point PA mutations demonstrated antagonistic effect on viral replication when combined with HA-226Q.Notably,any combination of PA mutations,along with double-point HA mutations,resulted in antagonistic effect on viral replication.We also observed antagonism in viral replication between PA-545V and PA-97I,as well as between HA-528V and PA-545V.Our findings demonstrated that several antagonistic mutations in HA and PA proteins affect viral replication,which may contribute to the H9N2 virus adaptation to mice and mammalian cells.These findings can potentially contribute to the monitoring of H9N2 field strains for assessing their potential risk in mammals.
基金supported by US National Institutes of Health grants Al147394,AG069264,Al112844,HL170961 and Al154598 to J.S.
文摘Pulmonary macrophages,such as tissue-resident alveolar and interstitial macrophages and recruited monocyte-derived macrophages,are the major macrophages present in the lungs during homeostasis and diseased conditions.While tissue-resident macrophages act as sentinels of the alveolar space and play an important role in maintaining homeostasis and immune regulation,recruited macrophages accumulate in the respiratory tract after acute viral infections.Despite sharing similar anatomical niches,these macrophages are distinct in terms of their origins,surface marker expression,and transcriptional profiles,which impart macrophages with distinguished characteristics in physi-ological and pathophysiological conditions.In this review,we summarize the current view on these macrophage populations,their shared functions,and what makes them distinct from each other in the context of homeostasis andrespiratoryviral infections.
文摘流行性感冒(简称“流感”)是由流感病毒引起的急性呼吸道传染疾病,据世界卫生组织统计,流感每年可导致300万~500万严重病例,其中29万~65万病例死亡,给社会带来沉重的经济负担,是一个世界性的公共卫生难题。研究发现宿主细胞中存在多条信号通路参与对流感病毒感染的应答,越来越多的研究表明宿主miRNAs通过直接或间接的方式,在流感病毒感染、复制的不同阶段发挥着重要调控作用。本文综合分析了目前关于宿主细胞miRNA对流感病毒复制调控的研究进展,对不同的miRNA具体的调控机制进行系统地归类总结后发现:甲型流感病毒(Influenza A virus,IAV)的PB1、PB2、NA、NP、M1基因是宿主miRNA直接抑制病毒复制的主要靶基因,而在间接调控过程中宿主miRNA主要作用在RIG-I样受体信号通路,Jak-STAT信号通路和Toll样受体信号通路三条流感病毒应答信号途径中,以上发现将更有助于全面理解宿主miRNA对于流感病毒调控网络和宿主细胞与流感病毒的互作机制。
文摘神经介素B(neuromedin B,NMB)及受体(NMB receptor,NMBR)通过NF-κB信号通路参与抑制甲型流感病毒(influenza A virus,IAV)H1N1亚型(IAV/H1N1)的感染。但关于NMB和NMBR对NF-κB信号通路相关泛素蛋白酶的调控如何,尚未见报道。为探究NMB和NMBR调控IAV诱导的NF-κB信号通路相关的泛素蛋白酶表达的影响,本研究基于IAV/H9N2感染sh-NMBR细胞与NMB刺激的A549细胞,采用RT-PCR、qRT-PCR及Western blot(WB)分析NMB和NMBR对H9N2感染引起的NF-κB信号通路相关的E3泛素连接酶Mind bomb-2(MIB2)和Ring Finger Protein 8(RNF8)及去泛素蛋白酶Cylindromatosis(CYLD)的表达变化。结果显示:H9N2感染sh-NMBR细胞中,RNF8和CYLD表达增加,MIB2表达和P65磷酸化水平降低;NMB激活A549细胞中NMBR的表达后,诱导细胞中RNF8和CYLD的表达水平下降,MIB2表达和P65磷酸化水平增加。结果表明:NMB和NMBR通过调控IAV/H9N2感染诱导的泛素蛋白酶的表达和P65活性,从而影响IAV/H9N2感染激活的NF-κB信号通路的功能,该结果为深入研究NMB和NMBR抑制甲型流感病毒感染的作用机制提供了理论基础。
