scholarly journals Generation and characterization of a monoclonal antibody against duck Tembusu virus envelope protein

2016 ◽  
Vol 19 (4) ◽  
pp. 877-883 ◽  
Author(s):  
K. Han ◽  
D. Zhao ◽  
Y. Liu ◽  
Q. Liu ◽  
X. Huang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Elisa Test ◽  
Antibody Detection ◽  
Economic Losses ◽  
Structural Protein ◽  
Neutralization Titer ◽  
Viral Attachment ◽  
E Protein ◽  
Duck Tembusu Virus ◽  
Tembusu Virus

Abstract Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. Envelope (E) protein of DTMUV is an important structural protein, which is able to induce protective immune response in target animals and can be used as specific serological diagnosis tool. In this study, a novel monoclonal antibody, designated mAb 3E9, was generated against DTMUV E protein. It is positive in indirect ELISA against both His-E protein and the purified whole viral antigen. Also, this mAb showed positive reaction with DTMUV in Western blot and indirect immunofluorescence assay, and the isotype was IgG1. End-point neutralizing assay performed in BHK-21 cells revealed that the neutralization titer of 3E9 against DTMUV JS804 strain reached 1:50. Furthermore, functional studies revealed that 3E9 blocks infection of DTMUV at a step on viral attachment. The anti-E mAbs produced in the present work may be valuable in developing an antigen-capture ELISA test for antigen detection or a competitive ELISA test for antibody detection or therapeutic medicine for DTMUV in poultry.

scholarly journals pUC18-CpG Is an Effective Adjuvant for a Duck Tembusu Virus Inactivated Vaccine

Viruses ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 238 ◽  
Author(s):  
Xiao Ren ◽  
Xiaolei Wang ◽  
Shan Zhang ◽  
Xintao Gao ◽  
Lichun Fang ◽  
...  
Keyword(s):  
Protective Immunity ◽  
Protective Efficacy ◽  
Vaccine Dose ◽  
Economic Losses ◽  
Control Group ◽  
Serum Cytokines ◽  
Duck Tembusu Virus ◽  
Antibody Titers ◽  
Protection Efficacy ◽  
Tembusu Virus

Duck Tembusu virus (DTMUV) is an emerging pathogenic flavivirus responsible for massive economic losses in the duck industry. However, commercially inactivated DTMUV vaccines have been ineffective at inducing protective immunity in ducks. The widely used adjuvant cytosine-phosphate-guanine oligodeoxynucleotides (CpG ODNs) reportedly improve humoral and cellular immunities in animal models. However, its effectiveness in DTMUV vaccines requires validation. Here, we assessed the protective efficacy of pUC18-CpG as an adjuvant in an inactivated live DTMUV vaccine in ducks. Our results revealed that the serum hemagglutination inhibition (HI) antibody titers, positive rates of anti-DTMUV antibodies, the concentration of serum cytokines, and protection efficacy were significantly increased in ducks immunized with pUC18-CpG compared to that in the control group. Moreover, ducks immunized with a full vaccine dose containing a half dose of antigen supplemented with 40 μg of pUC18-CpG exhibited the most potent responses. This study suggests that pUC18-CpG is a promising adjuvant against DTMUV, which might prove effective in treating other viral diseases in waterfowl.

scholarly journals Screening of Duck Tembusu Virus NS3 Interacting Host Proteins and Identification of Its Specific Interplay Domains

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 740 ◽  
Author(s):  
Yawen Wang ◽  
Shuai Zhang ◽  
Yi Tang ◽  
Youxiang Diao
Keyword(s):  
Mitogen Activated Protein Kinase ◽  
Structural Protein ◽  
Host Proteins ◽  
Host Immune Responses ◽  
Yeast Two Hybrid System ◽  
Duck Tembusu Virus ◽  
Ns3 Protein ◽  
Mitogen Activated Protein ◽  
Tembusu Virus ◽  
Information Database

NS3 protein is a member of the non-structural protein of duck Tembusu virus (DTMUV), which contains three domains, each of which has serine protease, nucleotide triphosphatase, and RNA helicase activities, respectively. It performs a variety of biological functions that are involved in the regulation of the viral life cycle and host immune response. Based on the yeast two-hybrid system, we successfully transformed pGBKT7-NS3 bait plasmid into Y2H Gold, tested it to prove that it has no self-activation and toxicity, and then hybridized it with the prey yeast strain of the duck embryo fibroblast cDNA library for screening. After high-stringency selection, positive alignment with the National Center for Biotechnology Information database revealed nine potential interactive proteins: MGST1, ERCC4, WIF1, WDR75, ACBD3, PRDX1, RPS7, ND5, and LDHA. The most interesting one (PRDX1) was selected to be verified with full-length NS3 protein and its three domains S7/DEXDc/HELICc using yeast regressive verification and GST Pull-Down assay. It denoted that PRDX1 does indeed interact with HELICc domains of NS3. NS3 is involved in the RNA uncoiling process of viral replication, which may cause mitochondrial overload to create oxidative stress (OS) during DTMUV attack. We deduced that the HELICc domain binding partner PRDX1, which regulates the p38/mitogen-activated protein kinase pathway (p38/MAPK) to avert OS, causing apoptosis, making it possible for viruses to escape host immune responses.

scholarly journals Adaptation and Attenuation of Duck Tembusu Virus Strain Du/CH/LSD/110128 following Serial Passage in Chicken Embryos

2014 ◽  
Vol 21 (8) ◽  
pp. 1046-1053 ◽  
Author(s):  
Ling Sun ◽  
Yunxia Li ◽  
Yue Zhang ◽  
Zongxi Han ◽  
Yang Xu ◽  
...  
Keyword(s):  
Virulent Strain ◽  
Serial Passage ◽  
Economic Losses ◽  
Suitable Candidate ◽  
Mortality And Morbidity ◽  
Viral Loads ◽  
Duck Tembusu Virus ◽  
Tembusu Virus ◽  
Chicken Eggs ◽  
Embryonated Chicken

ABSTRACTDuck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. In the current study, a virulent strain of DTMUV, designated Du/CH/LSD/110128, was isolated from the livers of diseased ducks and attenuated by serial passage in embryonated chicken eggs. The virus was partially attenuated after 50 and 70 passages and was fully attenuated after 90 passages, based on mortality and morbidity rates and viral loads in inoculated ducklings. Fourteen amino acid substitutions were observed in the capsid, prM, envelope, NS1, NS3, NS4A, NS4B, and NS5 proteins of the fully attenuated strain of Du/CH/LSD/110128, which might be responsible for the observed changes in replication and pathogenicity. A 72-nucleotide deletion was also observed in the 3′ untranslated region of the virus after 30 passages. The fully attenuated virus retained the immunogenicity of the parental strain, providing effective protection to challenge with virulent Du/CH/LSD/110128, and may represent a suitable candidate as a vaccine strain against DTMUV infection in ducks. Our results also lay the foundation for future studies on the replication and pathogenic mechanisms of DTMUV.

scholarly journals Identification of a New Broadly Cross-reactive Epitope within Domain III of the Duck Tembusu Virus E Protein

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Chenxi Li ◽  
Xiaofei Bai ◽  
Runze Meng ◽  
Wulin Shaozhou ◽  
Qingshan Zhang ◽  
...  
Keyword(s):  
E Protein ◽  
Duck Tembusu Virus ◽  
Tembusu Virus ◽  
Domain Iii

scholarly journals A recombinant adenovirus expressing the E protein of duck Tembusu virus induces protective immunity in duck

2019 ◽  
Vol 81 (2) ◽  
pp. 314-320 ◽  
Author(s):  
Jingyu TANG ◽  
Dongdong YIN ◽  
Rui WANG ◽  
Qi ZHOU ◽  
Xiaoya ZHOU ◽  
...  
Keyword(s):  
Protective Immunity ◽  
Recombinant Adenovirus ◽  
E Protein ◽  
Duck Tembusu Virus ◽  
Tembusu Virus

scholarly journals Basic Amino Acid Substitution at Residue 367 of the Envelope Protein of Tembusu Virus Plays a Critical Role in Pathogenesis

2020 ◽  
Vol 94 (8) ◽  
Author(s):  
Mengxu Sun ◽  
Lijiao Zhang ◽  
Yanxin Cao ◽  
Jun Wang ◽  
Ziding Yu ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Substitution ◽  
Critical Role ◽  
Basic Amino Acid ◽  
Site Directed Mutagenesis ◽  
Economic Losses ◽  
E Protein ◽  
Virulence Attenuation ◽  
Tembusu Virus

ABSTRACT Tembusu virus (TMUV) is a flavivirus responsible for panzootic outbreaks of severe egg-drop and fatal encephalitis of domestic waterfowl in China. Although TMUV can be attenuated by in vitro passaging, experimental evidence supporting the role of specific genetic changes in virulence attenuation is currently lacking. Here, we performed site-directed mutagenesis on five envelope (E) protein amino acid residues in accordance with the attenuated TMUV generated in our recent study. Our results showed that the Thr-to-Lys mutation of residue 367 in E protein (E367) plays a predominant role in viral cell adaptation and virulence attenuation in ducks compared with mutations in other residues. We further demonstrated that the positively charged basic amino acid substitution at E367 enhanced the viral binding affinity for glycosaminoglycans (GAGs) and reduced viremia levels and the efficiency of replication in major target organs in subcutaneously inoculated ducks. Interestingly, the T367K mutation increased viral neutralization sensitivity to the early immune sera. Together, our findings provide the first evidence that a basic amino acid substitution at E367 strongly impacts the in vitro and in vivo infection of TMUV. IMPORTANCE Outbreaks of Tembusu virus (TMUV) infection have caused huge economic losses in the production of domestic waterfowl since the virus was first recognized in China in 2010. To control TMUV infection, a live-attenuated vaccine candidate of TMUV was developed in our previous study, but the mechanisms of virulence attenuation are not fully understood. Here, we found that the Thr-to-Lys substitution at E367 is a crucial determinant of TMUV virulence attenuation in ducks. We demonstrated that the T367K mutation attenuates TMUV through reducing viral replication in the blood, brain, heart (ducklings), and ovaries. These data provide new insights into understanding the pathogenesis of TMUV and the rational development of novel TMUV vaccines.

scholarly journals ITRAQ-Based Quantitative Proteomics Reveals the Proteome Profiles of Primary Duck Embryo Fibroblast Cells Infected with Duck Tembusu Virus

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Feng Hu ◽  
Yufeng Li ◽  
Kexiang Yu ◽  
Bing Huang ◽  
Xiuli Ma ◽  
...  
Keyword(s):  
Absolute Quantification ◽  
Differentially Expressed ◽  
Economic Losses ◽  
Valuable Insight ◽  
Fibroblast Cells ◽  
Differentially Expressed Proteins ◽  
Duck Embryo Fibroblast ◽  
Duck Tembusu Virus ◽  
Duck Embryo ◽  
Tembusu Virus

Outbreaks of duck Tembusu virus (DTMUV) have caused substantial economic losses in the major duck-producing regions of China since 2010. To improve our understanding of the host cellular responses to virus infection and the pathogenesis of DTMUV infection, we applied isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with multidimensional liquid chromatography-tandem mass spectrometry to detect the protein changes in duck embryo fibroblast cells (DEFs) infected and mock-infected with DTMUV. In total, 434 cellular proteins were differentially expressed, among which 116, 76, and 339 proteins were differentially expressed in the DTMUV-infected DEFs at 12, 24, and 42 hours postinfection, respectively. The Gene Ontology analysis indicated that the biological processes of the differentially expressed proteins were primarily related to cellular processes, metabolic processes, biological regulation, response to stimulus, and cellular organismal processes and that the molecular functions in which the differentially expressed proteins were mainly involved were binding and catalytic activity. Some selected proteins that were found to be differentially expressed in DTMUV-infected DEFs were further confirmed by real-time PCR. The results of this study provide valuable insight into DTMUV-host interactions. This could lead to a better understanding of DTMUV infection mechanisms.

Liposomes containing recombinant E protein vaccine against duck Tembusu virus in ducks

Vaccine ◽  
2016 ◽  
Vol 34 (19) ◽  
pp. 2157-2163 ◽  
Author(s):  
Tengfei Ma ◽  
Yongxia Liu ◽  
Jia Cheng ◽  
Yanhan Liu ◽  
Wentao Fan ◽  
...  
Keyword(s):  
Protein Vaccine ◽  
E Protein ◽  
Duck Tembusu Virus ◽  
Tembusu Virus
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