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Emergence of H5N1 highly pathogenic avian influenza in Democratic People's Republic of Korea

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Journal of Integrative Agriculture 2022年第5期21卷 1534-1538页

作者： LIU Li-ling YANG Huan-liang GUO Fu-sheng WANG Xiu-rong DENG Guo-hua SHI Jian-zhong TIAN Guo-bin ZENG Xian-ying State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069P.R.China China Animal Health and Epidemiology Center Qingdao 266032P.R.China

In the past decade,there has been extensive global surveillance for highly pathogenic avian influenza(HPAI)infection in both animals and humans,however,few studies on epidemiology of avian influenza in Democratic People’s Republic of Korea(DPRK)were published.During the period 2013–2014,HPAI H5N1 viruses were detected with outbreaks in domestic poultry in DPRK.Phylogenetic analysis revealed that the hemagglutinin gene of all samples belonged to clade 2.***.2.1c with high homology.The HPAI H5N1 virus found in ducks at the Tudan Duck Farm in 2013 was might introduced by migratory birds and then led to the outbreaks on neighboring chicken farms in 2014.These data provide direct evidence for the transmission of avian influenza viruses from wild birds to waterfowl to terrestrial birds.Therefore,the monitoring and control of influenza virus in ducks must be given top priority,which are essential components to prevent and control HPAI.

关键词： avian influenza H5N1 Democratic People’s Republic of Korea clade 2.3.2.1c

Equine ANP32 proteins support influenza A virus RNA polymerase activity

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Virologica Sinica 2023年第6期38卷 951-960页

作者： Yuan Zhang Xing Guo Mengmeng Yu Liuke Sun Yuxing Qu Kui Guo Zhe Hu Diqiu Liu Haili Zhang Xiaojun Wang State Key Laboratory for Animal Disease Control and Prevention Harbin Veterinary Research InstituteThe Chinese Academy of Agricultural SciencesHarbin150069China State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases Key Laboratory for Zoonosis Research of the Ministry of EducationInstitute of Zoonosis and College of Veterinary MedicineJilin UniversityChangchun130062China

Host ANP32 family proteins are crucial for maintaining the activity of influenza RNA polymerase and play an important role in the cross-species transmission of influenza viruses.To date,the molecular properties of equine ANP32(eqANP32)protein are poorly understood,particularly the mechanisms that affect equine influenza virus(EIV)RNA polymerase activity.Here,we found that there are six alternative splicing variants of equine ANP32A(eqANP32A)with different levels of expression.Further studies showed that these six splicing variants of eqANP32A supported the activity of EIV RNA polymerase to varying degrees,with the variant eqANP32A_X2 having the highest expression abundance and exhibiting the highest support of polymerase activity.Sequence analysis demonstrated that the differences in the N-Cap regions of the six splicing variants significantly affected their N-terminal conformation,but did not affect their ability to bind RNA polymerase.We also demonstrated that there is only one transcript of eqANP32B,and that this transcript showed only very low support to the EIV RNA polymerase.This functional defect in eqANP32B is caused by the sequence of the 110–259 amino acids at its Cterminus.Our results indicated that it is the eqANP32A_X2 protein that mainly determines the efficiency of the EIV replication in horses.In conclusion,our study parsed the molecular properties of eqANP32 family proteins and revealed the sequence features of eqANP32A and eqANP32B,suggesting for the first time that the N-Cap region of ANP32A protein also plays an important role in supporting the activity of the influenza virus polymerase.

关键词： Equine influenza virus(EIV) Equine ANP32A Equine ANP32B RNA polymerase activity N-Cap domain

Generation and application of replication-competent Venus-expressing H5N1,H7N9,and H9N2 influenza A viruses

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Science Bulletin 2018年第3期63卷 176-186页

作者： guangwen wang jie zhang fandi kong qibing li jinliang wang shujie ma yuhui zhao libin liang junping li nan sun lizheng guan yuan zhou chenchen zhou shanyu huang zhigao bu li jiang hualan chen chengjun li State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences Harbin 150069 China

The generation and application of replication-competent influenza A virus(IAV) expressing a reporter gene represent a valuable tool to elucidate the mechanism of viral pathogenesis and establish new countermeasures to combat the threat of influenza. Here, replication-competent IAVs with a neuraminidase(NA) segment harboring a fluorescent reporter protein, Venus, were generated in the background of H5N1, H7N9, and H9N2 influenza viruses, the three subtypes of viruses with imminent pandemic potential. All three reporter viruses maintained virion morphology, replicated with similar or slightly reduced titers relative to their parental viruses, and stably expressed the fluorescent signal for at least two passages in embryonated chicken eggs. As a proof of concept, we demonstrated that these reporter viruses,used in combination with a high-content imaging system, can serve as a convenient and rapid tool for the screening of antivirals and host factors involved in the virus life cycle. Moreover, the reporter viruses demonstrated similar growth properties and tissue tropism as their parental viruses in mice, among which the H7N9 NA-Venus virus could potentially be used in ex vivo studies to better understand H7N9 pathogenesis or to develop novel therapeutics.

关键词：流行性感冒病毒 H9N2 维纳斯复制 H5N1 tial 种子类型联合使用

Acetyl-11-keto-beta-boswellic acid promotes sciatic nerve repair after injury: molecular mechanism

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Neural Regeneration research 2022年第12期17卷 2778-2784页

作者： Yao Wang Zong-Liang Xiong Xiang-Lin Ma Chong Zhou Mo-Han Huo Xiao-Wen Jiang Wen-Hui Yu Department of veterinary Medicine Northeast Agricultural UniversityHarbinHeilongjiang ProvinceChina College of Life Sciences Northeast Agricultural UniversityHarbinHeilongjiang ProvinceChina Key Laboratory of Heilongjiang Education Department for Common Animal Disease Prevention and Treatment Northeast Agricultural UniversityHarbinHeilongjiang ProvinceChina institute of Chinese veterinary Medicine Northeast Agricultural UniversityHarbinHeilongjiang ProvinceChina

Previous studies showed that acetyl-11-keto-beta-boswellic acid(AKBA),the active ingredient in the natural Chinese medicine Boswellia,can stimulate sciatic nerve injury repair via promoting Schwann cell proliferation.However,the underlying molecular mechanism remains poorly understood.In this study,we performed genomic sequencing in a rat model of sciatic nerve crush injury after gastric AKBA administration for 30 days.We found that the phagosome pathway was related to AKBA treatment,and brain-derived neurotrophic factor expression in the neurotrophic factor signaling pathway was also highly up-regulated.We further investigated gene and protein expression changes in the phagosome pathway and neurotrophic factor signaling pathway.Myeloperoxidase expression in the phagosome pathway was markedly decreased,and brain-derived neurotrophic factor,nerve growth factor,and nerve growth factor receptor expression levels in the neurotrophic factor signaling pathway were greatly increased.Additionally,expression levels of the inflammatory factors CD68,interleukin-1β,pro-interleukin-1β,and tumor necrosis factor-αwere also decreased.Myelin basic protein-andβ3-tubulin-positive expression as well as the axon diameter-to-total nerve diameter ratio in the injured sciatic nerve were also increased.These findings suggest that,at the molecular level,AKBA can increase neurotrophic factor expression through inhibiting myeloperoxidase expression and reducing inflammatory reactions,which could promote myelin sheath and axon regeneration in the injured sciatic nerve.

关键词： AKBA axon genomics inflammatory injury and repair myelin sheath myeloperoxidase neurotrophic factor peripheral nerve phagosome pathway regeneration Sprague-Dawley rat

ASFV transcription reporter screening system identifies ailanthone as a broad antiviral compound

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Virologica Sinica 2023年第3期38卷 459-469页

作者： Yuhang Zhang Zhenjiang Zhang Fan Zhang Jiwen Zhang Jun Jiao Min Hou Nianchao Qian Dongming Zhao Xiaofeng Zheng Xu Tan College of veterinary Medicine Hunan Agricultural UniversityChangsha 410128China State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150009China Beijing Advanced Innovation Center for Structural Biology Beijing Frontier Research Center for Biological StructureMOE Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical BiologySchool of Pharmaceutical SciencesTsinghua-Peking Center for Life SciencesTsinghua UniversityBeijing 100084China institute of Health and Medicine Hefei Comprehensive National Science CenterHefei 230601 China

African swine fever(ASF)is an acute,highly contagious and deadly viral disease in swine that jeopardizes the worldwide pig industry.Unfortunately,there are no authoritative vaccine and antiviral drug available for ASF control.African swine fever virus(ASFV)is the etiological agent of ASF.Among the ASFV proteins,p72 is the most abundant component in the virions and thus a potential target for anti-ASFV drug design.Here,we con-structed a luciferase reporter system driven by the promoter of p72,which is transcribed by the co-transfected ASFV RNA polymerase complex.Using this system,we screened over 3200 natural product compounds and obtained three potent candidates against ASFV.We further evaluated the anti-ASFV effects and proved that among the three candidates,ailanthone(AIL)inhibits the replication of ASFV at the nanomolar concentration(IC_(50)=15 nmol/L).Our in vitro experiments indicated that the antiviral effect of AIL is associated with its inhibition of the HSP90-p23 cochaperone.Finally,we showed the antiviral activity of AIL on Zika virus and hepatitis B virus(HBV),which supports that AIL is a potential broad-spectrum antiviral agent.

关键词： African swine fever virus(ASFV) Replication inhibition Broad-spectrum antiviral drugs Ailanthone(AIL)

Novel P22-monoclonal antibody based blocking ELISA for the detection of African swine fever virus antibodies in serum

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Biosafety and Health 2022年第4期4卷 234-243页

作者： Ghebremedhin Tsegay Weldu Tesfagaber Yuanmao Zhu Xijun He Wan Wang Zhenjiang Zhang Encheng Sun Jinya Zhang Yuntao Guan Fang Li Renqiang Liu Zhigao Bu Dongming Zhao State Key Laboratory of veterinary Biotechnology National High Containment Facilities for Animal Diseases Control and PreventionNational African Swine Fever Para-reference LaboratoryHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China Department of veterinary Science Hamelmalo Agricultural CollegeKerenEritrea

African swine fever(ASF)is a highly infectious,transboundary viral disease of domestic and wild pigs,and is currently the most serious threat to world swine production,resulting in significant economic loss.In the absence of vaccines and treatments,the control of the disease entirely depends on accurate and early diagnosis accompanied by the culling of infected pigs.Thus,a highly specific and sensitive diagnostic assay is required during an outbreak and surveillance of the disease.In this study,a highly sensitive,specific,rapid and repeatable P22-monoclonal antibody-based blocking enzyme-linked immunosorbent assay(bELISA)assay was developed for the detection of antibodies against genotype I and II African swine fever viruses(ASFVs).A total of 806 pig serum samples were tested to evaluate the performance of the diagnostic assay.To determine the PI(percent Inhibition)cut-off value,receiver-operating characteristic(ROC)analysis was applied.According to the ROC analysis of the data,98.10%specificity and 100%sensitivity were recorded when the threshold cut-off value of PI was established at 47%.In addition,the assay was able to detect ASFV antibodies as early as 9 days post-infection when serum samples from experimentally infected pigs were used.Taking all together,the results of the present study indicated that the P22-mAb based bELISA assay can be used for rapid and accurate detection of antibodies against ASFV,which could play a valuable role in the containment and prevention of ASFV as an alternative to other serological diagnostic methods.Also,this study will assist researchers to further investigate the immunogenic importance of P22 protein in ASFV infection.

关键词： Monoclonal antibodies African swine fever Blocking ELISA Diagnosis P22

Vaccination of poultry successfully eliminated human infection with H7N9 virus in China

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Science China(Life Sciences) 2018年第12期61卷 1465-1473页

作者： Xianying Zeng Guobin Tian Jianzhong Shi Guohua Deng Chengjun Li Hualan Chen State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China

The H7 N9 viruses that emerged in China in 2013 were nonpathogenic in chickens but mutated to a highly pathogenic form in early 2017 and caused severe disease outbreaks in chickens. The H7 N9 influenza viruses have caused five waves of human infection, with almost half of the total number of human cases(766 of 1,567) being reported in the fifth wave, raising concerns that even more human infections could occur in the sixth wave. In September 2017, an H5/H7 bivalent inactivated vaccine for chickens was introduced, and the H7 N9 virus isolation rate in poultry dropped by 93.3% after vaccination. More importantly,only three H7 N9 human cases were reported between October 1, 2017 and September 30, 2018, indicating that vaccination of poultry successfully eliminated human infection with H7 N9 virus. These facts emphasize that active control of animal disease is extremely important for zoonosis control and human health protection.

关键词： H7N9 influenza virus evolution vaccination human infection elimination

Next-generation sequencing and single-cell RT-PCR reveal a distinct variable gene usage of porcine antibody repertoire following PEDV vaccination

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Science China(Life Sciences) 2020年第8期63卷 1240-1250页

作者： Ren Li Fang Fu Li Feng PingHuang Liu State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150001China

Porcine epidemic diarrhea virus(PEDV)is the most common diarrhea-causing pathogen in newborn piglets.The clarifications of the overall antibody repertoire and antigen-specific antibody repertoire are essential to provide important insights into the B-cell response and reshape new vaccines.Here,we applied next-generation sequencing(NGS)technology to investigate immunoglobulin(Ig)variable(V)gene segment usage of swine B-cells from peripheral blood lymphocytes(PBL)and mesenteric lymph node(MLN)cells following PEDV vaccination.We identified the transcripts of all functional Ig V-genes in antibody repertoire.IgHV1 S2,IgKV1-11,and IgLV3-4 were the most prevalent gene segments for heavy,kappa,and lambda chains,respectively,in PBL and MLN.Unlike previous studies,IgKV1,instead of IgKV2,and IgLV3,instead of IgLV8,were the prevalent Ig V-gene families for kappa and lambda light chains,respectively.We further examined the antibody repertoire of PEDV spike-specific B cells by single-cell RT-PCR.In contrast to the overall antibody repertoire,Ig V-gene segments of PEDV spike-specific B cells preferentially adopted IgHV1-4 and IgHV1-14 for heavy chain,IgKV1-11 for kappa chain,and IgLV3-3 for lambda chain.These results represent a comprehensive analysis to characterize the Ig V-gene segment usage in the overall and PEDV spike-specific antibody repertoire in PBL and MLN.

关键词： porcine epidemic diarrhea virus(PEDV) next-generation sequencing(NGS) single-cell RT-PCR antibody repertoire

TRIM35 mediates protection against influenza infection by activating TRAF3 and degrading viral PB2

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Protein & Cell 2020年第12期11卷 894-914页

作者： Nan Sun Li Jiang Miaomiao Ye Yihan Wang Guangwen Wang Xiaopeng Wan Yuhui Zhao Xia Wen Libin Liang Shujie Ma Liling Liu Zhigao Bu Hualan Chen Chengjun Li State Key Laboratory of veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China

Tripartite motif(TRIM)family proteins are important effectors of innate immunity against viral infections.Here we identified TRIM35 as a regulator of TRAF3 activation.Deficiency in or inhibition of TRIM35 suppressed the production of type I interferon(IFN)in response to viral infection.777m35-deficient mice were more susceptible to influenza A virus(IAV)infection than were wild-type mice.TRIM35 promoted the RIG-Imediated signaling by catalyzing Lys63-linked polyubiquitination of TRAF3 and the subsequent formation of a signaling complex with VISA and TBK1.IAV PB2 polymerase countered the innate antiviral immune response by impeding the Lys63-linked polyubiquitination and activation of TRAF3.TRIM35 mediated Lys48-linked polyubiquitination and proteasomal degradation of IAV PB2,thereby antagonizing its suppression of TRAF3 activation.Our in vitro and in vivo findings thus reveal novel roles of TRIM35,through catalyzing Lys63-or Lys48-linked polyubiquitination,in RIG-I antiviral immunity and mechanism of defense against IAV infection.

关键词： influenza A virus PB2 TRIM35 TRAF3 ubiquitination antiviral immunity