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Ferroptosis contributes to JEV-induced neuronal damage and neuroinflammation

Virologica Sinica 2024年第1期39卷 144-155页

作者： Wenjing Zhu Qi Li Yong Yin Huanchun Chen Youhui Si Bibo Zhu shengbo cao Zikai Zhao Jing Ye National Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan430070China Frontiers Science Center for Animal Breeding and Sustainable Production Huazhong Agricultural UniversityWuhan430070China The Cooperative Innovation Center for Sustainable Pig Production Huazhong Agricultural UniversityWuhan430070China Hubei Hongshan Laboratory Wuhan430070China

Ferroptosis is a newly discovered prototype of programmed cell death (PCD) driven by iron-dependent phospholipid peroxidation accumulation, and it has been linked to numerous organ injuries and degenerative pathologies. Although studies have shown that a variety of cell death processes contribute to JEV-induced neuroinflammation and neuronal injury, there is currently limited research on the specific involvement of ferroptosis. In this study, we explored the neuronal ferroptosis induced by JEV infection in vitro and in vivo. Our results indicated that JEV infection induces neuronal ferroptosis through inhibiting the function of the antioxidant system mediated by glutathione (GSH)/glutathione peroxidase 4 (GPX4), as well as by promoting lipid peroxidation mediated by yes-associated protein 1 (YAP1)/long-chain acyl-CoA synthetase 4 (ACSL4). Further analyses revealed that JEV E and prM proteins function as agonists, inducing ferroptosis. Moreover, we found that treatment with a ferroptosis inhibitor in JEV-infected mice reduces the viral titers and inflammation in the mouse brains, ultimately improving the survival rate of infected mice. In conclusion, our study unveils a critical role of ferroptosis in the pathogenesis of JEV, providing new ideas for the prevention and treatment of viral encephalitis.

关键词： Japanese encephalitisvirus(JEV) Ferroptosis Neuron Lipid peroxidation Inflammatory response

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猫IL-1β阻断单克隆抗体的筛选与猫源化改造

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中国兽医科学 2024年第3期54卷 364-372页

作者：王定一王阅张晨曦李秋燕周登元司有辉曹胜波叶静华中农业大学动物医学院湖北武汉430070

为给猫的炎症性疾病治疗提供具有与猫白细胞介素-1β(IL-1β)高亲和力的单克隆抗体,将表达后纯化的重组猫IL-1β蛋白接种小鼠后制备单克隆抗体,经验证单抗对猫IL-1β蛋白的阻断效果并分析其靶向位点后,将小鼠单克隆抗体的轻/重链可变区... 详细信息

为给猫的炎症性疾病治疗提供具有与猫白细胞介素-1β(IL-1β)高亲和力的单克隆抗体,将表达后纯化的重组猫IL-1β蛋白接种小鼠后制备单克隆抗体,经验证单抗对猫IL-1β蛋白的阻断效果并分析其靶向位点后,将小鼠单克隆抗体的轻/重链可变区基因片段连接到表达猫抗体的真核质粒中,转染真核细胞后检测猫源化改造抗体效果。结果表明,成功扩增了猫IL-1β基因,并成功构建了原核表达质粒pET-30a-IL-1β并表达猫IL-1β重组蛋白;在使用猫IL-1β重组蛋白免疫小鼠以及细胞融合、杂交瘤细胞筛选后,成功得到2株分别能够与猫IL-1β重组蛋白第二、第三结构域反应的小鼠源单克隆抗体C4C、H10E;将单克隆抗体C4C、H10E猫源化基因改造后的重组抗体仍具有阻断效果。本试验研究结果为开发针对猫IL-1β的阻断单克隆抗体药物奠定了基础,也为猫炎症性疾病的治疗提供了一种新的方案。

关键词：白介素1β 单克隆抗体阻断效果猫源化抗体改造

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猫白血病病毒gp70蛋白单克隆抗体的制备及应用

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中国兽医学报 2024年第1期44卷 74-79页

作者：杜文广彭佳佳曾悦李佳康史开拓周登元曹胜波张燕李秋燕华中农业大学动物医学院湖北武汉430070 武汉科前生物股份有限公司湖北武汉430040

猫白血病病毒(FeLV)是一种在家猫和野生猫科动物中水平传播的逆转录病毒,可引起淋巴肿瘤、免疫抑制、贫血等疾病,且国内缺乏针对FeLV的诊断试剂及预防或治疗用生物制剂。膜表面糖蛋白gp70是FeLV中和抗体的主要作用靶点,本研究通过构建g... 详细信息

猫白血病病毒(FeLV)是一种在家猫和野生猫科动物中水平传播的逆转录病毒,可引起淋巴肿瘤、免疫抑制、贫血等疾病,且国内缺乏针对FeLV的诊断试剂及预防或治疗用生物制剂。膜表面糖蛋白gp70是FeLV中和抗体的主要作用靶点,本研究通过构建gp70重组表达载体,表达纯化gp70重组蛋白,并以纯化的gp70蛋白免疫6周龄的BALB/c小鼠,进行单克隆抗体的制备。结果表明,成功构建了pET-32a-gp70重组表达质粒,并应用IPTG诱导表达、包涵体纯化获得了原核表达的gp70重组蛋白;成功制备出3株能稳定分泌针对gp70蛋白的单克隆抗体杂交瘤细胞株,分别命名为2H5、3B1、5A8;Western blot检测表明仅有2H5单克隆抗体能与gp70蛋白特异性结合;IFA检测表明2H5单克隆抗体能与真核杆状病毒表达的gp70蛋白发生特异性反应,2H5单抗效价可达1∶2048000。本研究成功制备了针对FeLV gp70蛋白的单克隆抗体,为进一步建立准确快速的FeLV诊断方法及研究预防或治疗用生物制剂奠定基础。

关键词：猫白血病病毒(FeLV) gp70蛋白原核表达单克隆抗体杆状病毒表达系统

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Epidemiological trends of mosquito-borne viral diseases in Pakistan

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Animal Diseases 2022年第2期2卷 127-136页

作者： Muhammad Imran Jing Ye Muhammad K.Saleemi Iqra Shaheen Ali Zohaib Zheng Chen shengbo cao State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan 430070HubeiChina Key Laboratory of Preventive Veterinary Medicine in Hubei Province College of Veterinary MedicineHuazhong Agricultural UniversityWuhan 430070HubeiChina The Cooperative Innovation Center for Sustainable Pig Production Huazhong Agricultural UniversityWuhan 430070HubeiChina Department of Pathology Faculty of Veterinary ScienceUniversity of AgricultureFaisalabad 38040Pakistan Livestock and Dairy Development Department PunjabPakistan Department of Microbiology Faculty of Veterinary&Animal SciencesThe Islamia University of BahawalpurBahawalpur 63100Pakistan Department of Preventive Veterinary Medicine College of Animal Science and TechnologyJiangxi Agricultural UniversityNanchang 330045JiangxiChina

Globally,arboviruses are public health *** has seen a fast-paced increase in mosquito-borne Flavivirus diseases such as dengue because of deforestation,climate change,urbanization,poor sanitation and natural *** magnitude and distribution of these diseases are poorly understood due to the lack of a competitive nationwide surveillance *** dengue-endemic countries,the recent epidemics of chikungunya(CHIKV)and human West Nile virus(WNV)have created panic among the public and are thought to provoke an outbreak of Zika virus(ZIKV)in ***,hospital-based surveillance has indicated the presence of Japanese encephalitis virus(JEV),which is deeply concerned by developing countries such as *** situation could become more devastating because of poorly developed diagnostic *** date,no licensed vaccine has been used in Pakistan,and preventive measures are mainly based on vector *** review provides comprehensive information concerning the association of risk factors with disease occurrence,epidemiological trends,and prediction of the spread of mosquito-borne diseases,attention to new threats of ZIKV,and future perspectives by benchmarking global health policies.

关键词： Arboviruses Dengue CHIKV JEV WNV ZIKA Public health

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Screening of novel synthetic derivatives of dehydroepiandrosterone for antivirals against flaviviruses infections

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Virologica Sinica 2022年第1期37卷 94-106页

作者： Muhammad Imran Luping Zhang Bohan Zheng Zikai Zhao Dengyuan Zhou Shengfeng Wan Zheng Chen Hongyu Duan Qiuyan Li Xueqin Liu shengbo cao Shaoyong Ke Jing Ye State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan430070China Key Laboratory of Preventive Veterinary Medicine in Hubei Province College of Veterinary MedicineHuazhong Agricultural UniversityWuhan430070China The Cooperative Innovation Center for Sustainable Pig Production Huazhong Agricultural UniversityWuhan430070China Department of Pathology Faculty of Veterinary ScienceUniversity of AgricultureFaisalabad38040Pakistan Department of Nephrology Henan Provincial Key Laboratory of Kidney Diseases and ImmunologyHenan Provincial People's HospitalZhengzhou450003China Key Laboratory for Animal Health of Jiangxi Province Nanchang330045China Department of Preventive Veterinary Medicine College of Animal Science and TechnologyJiangxi Agricultural UniversityNanchang330045China College of Fisheries Huazhong Agricultural UniversityWuhan430070China National Biopesticide Engineering Research Center Hubei Biopesticide Engineering Research CenterHubei Academy of Agricultural SciencesWuhan430070China

Flaviviruses are important arthropod-borne pathogens that represent an immense global health *** unprecedented epidemic rate and unpredictable clinical features underscore an urgent need for antiviral ***(DHEA)is a natural occurring adrenal-derived steroid in the human body that has been associated in protection against various *** the present study,the plaque assay based primary screening was conducted on 32 synthetic derivatives of DHEA against Japanese encephalitis virus(JEV)to identify potent anti-flaviviral *** on primary screening,HAAS-AV3026 and HAAS-AV3027 were selected as hits from DHEA derivatives that exhibited strong antiviral activity against JEV(IC_(50)=2.13 and 1.98μmol/L,respectively)and Zika virus(ZIKV)(IC_(50)=3.73 and 3.42μmol/L,respectively).Mechanism study indicates that HAAS-AV3026 and HAAS-AV3027 do not exhibit inhibitory effect on flavivirus binding and entry process,while significantly inhibit flavivirus infection at the replication ***,indirect immunofluorescence assay,Western blot analyses,and quantitative reverse transcription-PCR(qRT-PCR)revealed a potent antiviral activity of DHEA derivatives hits against JEV and ZIKV in terms of inhibition of viral infection,protein production,and viral RNA synthesis in Vero *** together,our results may provide a basis for the development of new antivirals against flaviviruses.

关键词： Flaviviruses Japanese encephalitis virus(JEV) Zika virus(ZIKV) Dehydroepiandrosterone(DHEA) Antivirals

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Isolation and phylogenetic analysis of feline calicivirus strains from various region of China

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Animal Diseases 2022年第3期2卷 165-174页

作者： Longlong cao Qiuyan Li Kaituo Shi Liting Wei Hehao Ouyang Zijun Ye Wenguang Du Jiawen Ye Xiaochen Hui Jiakang Li shengbo cao Dengyuan Zhou State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhanChina College of Veterinary Medicine Huazhong Agricultural UniversityWuhanChina Wuhan Keqian Biology Co. LtdWuhanChina

Feline calicivirus(FCV)is an important feline pathogen mainly causing upper respiratory tract disease,conjunctivitis,and stomatitis,and it is classifed into genotype I and genotype *** investigate the prevalence and molecular characteristics of FCV,this study collected 337 cat swab samples from animal hospitals in diferent regions of China from 2019 to *** positive detection rate of FCV was 29.9%(101/337)by *** analysis showed that FCV prevalence was signifcantly associated with living environment(p=0.0004),age(p=0.031)and clinical symptoms(p=0.00),but not with sex(p=0.092)and breed(p=0.171).The 26 strains of FCV were isolated using F81 *** analysis showed that 10 isolates belonged to genotype I,and 16 isolates belonged to genotype *** 26 isolates were highly genetically diverse,of which HB7 isolate had three same virulence-related amino acid loci with VSD *** loci distinguishing diferent genotypes were identifed from 26 isolates,suggesting the genetic relationship between diferent *** addition,selection pressure analysis based on capsid protein of 26 isolates revealed that the protein is under diversifying *** study reveals the genetic diversity of FCV and provides a reference for the screening of vaccine candidate strains and the development of vaccines with better cross-protection efects.

关键词： Feline Calicivirus Genetic diversity Phylogenetic analysis Selection pressure analysis ORF2 gene

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3D bioprinting of in vitro porous hepatoma models:establishment,evaluation,and anticancer drug testing

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Bio-Design and Manufacturing 2024年第2期7卷 137-152页

作者： Xiaoyuan Wang Zixian Liu Qianqian Duan Boye Zhang Yanyan cao Zhizhong Shen Meng Li Yanfeng Xi Jianming Wang shengbo Sang Shanxi Key Laboratory of Micro Nano Sensors and Artificial Intelligence Perception College of Electronic Information and Optical EngineeringTaiyuan University of TechnologyTaiyuan 030024China Key Lab of Advanced Transducers and Intelligent Control System of the Ministry of Education Taiyuan University of TechnologyTaiyuan 030024China Shanxi Institute of 6D Artificial Intelligence Biomedical Science Taiyuan 030031China Shanxi-Zheda Institute of Advanced Materials and Chemical Engineering Taiyuan 030024China Department of Pathology Shanxi Provincial Cancer HospitalTaiyuan 030024China General Hospital of Taiyuan Iron and Steel Company Taiyuan 030809China

Traditional tumor models do not tend to accurately simulate tumor growth in vitro or enable personalized treatment and are particularly unable to discover more beneficial targeted *** address this,this study describes the use of threedimensional(3D)bioprinting technology to construct a 3D model with human hepatocarcinoma SMMC-7721 cells(3DP-7721)by combining gelatin methacrylate(GelMA)and poly(ethylene oxide)(PEO)as two immiscible aqueous phases to form a bioink and innovatively applying fluorescent carbon quantum dots for long-term tracking of *** GelMA(10%,mass fraction)and PEO(1.6%,mass fraction)hydrogel with 3:1 volume ratio offered distinct pore-forming characteristics,satisfactorymechanical properties,and biocompatibility for the creation of the 3DP-7721 *** analysis and quantitative real-time fluorescence polymerase chain reaction(PCR)were used to evaluate the biological properties of the *** with the two-dimensional culture cell model(2D-7721)and the 3D mixed culture cell model(3DM-7721),3DP-7721 significantly improved the proliferation of cells and expression of tumor-related proteins and ***,we evaluated the differences between the three culture models and the effectiveness of antitumor drugs in the three models and discovered that the efficacy of antitumor drugs varied because of significant differences in resistance proteins and genes between the three *** addition,the comparison of tumor formation in the three models found that the cells cultured by the 3DP-7721 model had strong tumorigenicity in nude *** evaluation of the levels of biochemical indicators related to the formation of solid tumors showed that the 3DP-7721 model group exhibited pathological characteristics of malignant tumors,the generated solid tumors were similar to actual tumors,and the deterioration was *** research therefore acts as a foundation for the application of 3DP-7721 models in drug devel

关键词： 3D bioprinting Hepatoma tumor models Drug screening Antitumor drug development

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PRRSV、SS和Pm多重TaqMan荧光定量PCR检测方法的建立与初步应用

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中国兽医学报 2024年第2期44卷 231-236页

作者：詹存林王庚孙秀秀冯贺龙林正丹胡薛英谷长勤张万坡陶攀陈品钱平曹胜波张伟超程国富华中农业大学动物医学院湖北武汉430000 广西扬翔股份有限公司广西贵港537000

为了建立一种可同时检测并鉴别猪繁殖和呼吸综合征病毒(PRRSV)、猪链球菌(SS)和猪多杀性巴氏杆菌(Pm)多重TaqMan荧光定量PCR检测方法,本试验通过MEGA7.0分析NCBI上已登录的3种病原体基因序列,分别根据PRRSV的ORF6基因、SS的GDH基因和Pm... 详细信息

为了建立一种可同时检测并鉴别猪繁殖和呼吸综合征病毒(PRRSV)、猪链球菌(SS)和猪多杀性巴氏杆菌(Pm)多重TaqMan荧光定量PCR检测方法,本试验通过MEGA7.0分析NCBI上已登录的3种病原体基因序列,分别根据PRRSV的ORF6基因、SS的GDH基因和Pm的KMT1基因设计出特异性引物和探针,并对反应条件进行优化。结果显示,该方法能够特异性地检测PRRSV、SS和Pm,对其他病原无交叉反应;对PRRSV、SS和Pm的最低检测浓度分别为9.93×10^(2)、1.10×10^(2)和1.07×10^(2)拷贝/μL;组内和组间变异系数均小于1.85%。结果表明,该方法特异性强、灵敏性高、稳定性好,可应用于临床检测。

关键词：猪繁殖和呼吸综合征病毒猪链球菌猪多杀性巴氏杆菌多重TaqMan荧光定量PCR

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PD1^+CCR2^+CD8^+T Cells Infiltrate the Central Nervous System during Acute Japanese Encephalitis Virus Infection

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Virologica Sinica 2019年第5期34卷 538-548页

作者： Fang Zhang Linlin Qi Tong Li Xiaojing Li Dan Yang shengbo cao Jing Ye Bin Wei State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of Chinese Academy of Science Beijing 100049China State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhan 430070China School of Life Sciences Shanghai UniversityShanghai 200444China

Japanese encephalitis(JE)is a viral encephalitis disease caused by Japanese encephalitis virus(JEV)*** inflammatory responses in the central nervous system(CNS)are a hallmark of severe *** the CCR2-CCL2 axis is important for monocytes trafficking during JEV infection,little is known about its role in CNS trafficking of CD8^+T ***,we characterized a mouse model of JEV infection,induced via intravenous injection(i.v.)and delineated the chemokines and infiltrating peripheral immune cells in the brains of infected *** CNS expression of chemokines,Ccl2,Ccl3,and Ccl5,and their receptors,Ccr2 or *** significantly up-regulated after JEV infection and was associated with the degree of JE ***,JEV infection resulted in the migration of a large number of CD8^+T cells into the *** the brains of JEV-infected mice,infiltrating CD8^+T cells expressed CCR2 and CCR5 and were found to comprise mainly effector T cells(CD44^+CD62L_).JEV infection dramatically enhanced the expression of programmed death 1(PD-1)on infiltrating CD8^+T cells in the brain,as compared to that on peripheral CD8^+T cells in the *** effect was more pronounced on infiltrating CCR2^+CD8^+T cells than on CCR2-CD8^+T *** conclusion,we identified a new subset of CD8^+T cells(PD1^+CCR2^+CD8^+T cells)present in the CNS of mice during acute JEV *** CD8^+T cells might play a role in JE pathogenesis.

关键词： Japanese encephalitis virus(JEV) CD8^+T cell CCL2 CCR2 PD-1

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Japanese Encephalitis Virus NS1' Protein Antagonizes Interferon Beta Production

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Virologica Sinica 2018年第6期33卷 515-523页

作者： Dengyuan Zhou Fan Jia Qiuyan Li Luping Zhang Zheng Chen Zikai Zhao Min Cui Yunfeng Song Huanchun Chen shengbo cao Jing Ye State Key Laboratory of Agricultural Microbiology Huazhong Agricultural University Key Laboratory of Preventive Veterinary Medicine in Hubei Province College of Veterinary Medicine Huazhong Agricultural University The Cooperative Innovation Center for Sustainable Pig Production Huazhong Agricultural University Wuhan Institute of Physics and Mathematics (WIPM) Chinese Academy of Sciences

Japanese encephalitis virus(JEV) is a mosquito-borne virus and the major cause of viral encephalitis in Asia. NS1', a52-amino acid C-terminal extension of NS1, is generated with a-1 programmed ribosomal frameshift and is only present in members of the Japanese encephalitis serogroup of flaviviruses. Previous studies demonstrated that NS1' plays a vital role in virulence, but the mechanism is unclear. In this study, an NS1' defected(rG66A) virus was generated. We found that rG66A virus was less virulent than its parent virus(pSA14) in wild-type mice. However, similar mortality caused by the two viruses was observed in an IFNAR knockout mouse model. Moreover, we found that rG66A virus induced a greater type Ⅰ interferon(IFN) response than that by pSA14, and JEV NS1' significantly inhibited the production of IFN-b and IFN-stimulated genes. Taken together, our results reveal that NS1' plays a vital role in blocking type I IFN production to help JEV evade antiviral immunity and benefit viral replication.

关键词： Japanese encephalitis virus(JEV) NS1’ Type Ⅰ interferon(IFN-Ⅰ) Immune evasion

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