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Immunogenicity of a recombinant VSV-Vectored SARS-CoV vaccine induced robust immunity in rhesus monkeys after single-dose immunization

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Virologica Sinica 2022年第2期37卷 248-255页

作者： Dan Shan Xiaoyan Tang Renqiang Liu Dan Pan Xijun Wang Jinying Ge Zhiyuan Wen Zhigao Bu State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin150069China Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses Yangzhou UniversityYangzhou225127China

Severe acute respiratory syndrome(SARS)is a highly contagious zoonotic disease caused by SARS coronavirus(SARS-Co V).Since its outbreak in Guangdong Province of China in 2002,SARS has caused 8096 infections and774 deaths by December 31st,2003.Although there have been no more SARS cases reported in human populations since 2004,the recent emergence of a novel coronavirus disease(COVID-19)indicates the potential of the recurrence of SARS and other coronavirus disease among humans.Thus,developing a rapid response SARS vaccine to provide protection for human populations is still needed.Spike(S)protein of SARS-Co V can induce neutralizing antibodies,which is a pivotal immunogenic antigen for vaccine development.Here we constructed a recombinant chimeric vesicular stomatitis virus(VSV)VSVΔG-SARS,in which the glycoprotein(G)gene is replaced with the SARS-Co V S gene.VSVΔG-SARS maintains the bullet-like shape of the native VSV,with the heterogeneous S protein incorporated into its surface instead of G protein.The results of safety trials revealed that VSVΔG-SARS is safe and effective in mice at a dose of 1×10^(6)TCID_(50).More importantly,only a single-dose immunization of 2×10^(7)TCID_(50)can provide high-level neutralizing antibodies and robust T cell responses to non-human primate animal models.Thus,our data indicate that VSVΔG-SARS can be used as a rapid response vaccine candidate.Our study on the recombinant VSV-vectored SARS-Co V vaccines can accumulate experience and provide a foundation for the new coronavirus disease in the future.

关键词： Severe acute respiratory syndrome coronavirus(SARS-CoV) Vesicular stomatitis virus(VSV) Vaccine Immunization

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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

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Host protein ABCE1 interacts with the viral phosphoprotein and promotes rabies virus replication

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Biosafety and Health 2020年第3期2卷 157-163页

作者： Xing Liu Jiwen Zhang Fang Li Yibrah Tekle Hagoss Weldu Tesfagaber Lulu Wang Zilong Wang Dongming Zhao Zhigao Bu State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China National High Containment Laboratory for Animal Diseases Control and Prevention Harbin 150069China Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonosis Yangzhou UniversityYangzhou 225009China

Rabies virus(RABV)phosphoprotein(P)plays an important role in disrupting host interferon(IFN)-mediated antiviral immune response.ABCE1 is known as an RNase L inhibitor that negatively regulates the 2′,5′-oligoadenylate(2-5A)/RNase L antiviral system related to the IFN signaling pathway.In this study,we screened the host factors associated with RABV P protein,while focusing on ABCE1 because of its role in the life cycle of several viruses.Our results showed that knockout of ABCE1 in HEK293T cells inhibited RABV replication.In contrast,the overexpression of ABCE1 in HEK293T cells enhanced RABV replication.Notably,the co-immunoprecipitation assay showed that ABCE1 interacted with RABV P protein.Since ABCE1 and RABV P proteins are related to the IFN signaling pathway,we propose that the interaction of viral P protein with ABCE1 leads to the disruption of host antiviral immune response.In summary,our research showed that ABCE1 is an important host protein that interacts with viral P protein and regulates RABV replication.Moreover,the interaction between ABCE1 and RABV P protein may facilitate the viral P protein-mediated disruption of host antiviral immune response.

关键词： Rabies virus ABCE1 Replication Phosphoprotein Interaction

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Emergence of a Novel Bluetongue Virus Serotype in Xinjiang, China

Emergence of a Novel Bluetongue Virus Serotype in Xinjiang, ...

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第十届全国免疫学学术大会

作者： Encheng Sun Liping Huang Qingyuan Xu Zhigao Bu Donglia Wu Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences

Background:Bluetongue virus(BTV)belongs to the genus Orbivirus and infects ruminants,camelids and occasionally large carnivores.The recent identification of BTV-25,BTV-26,and BTV-27 in Switzerland,Kuwait and France,respectively,suggests that additional BTV serotypes may circulate around the world.Objective:Through conventional epidemiological investigations confirmed epidemiological situation of BTV in Xinjiang,China.Methods:Conventional methods were used to isolate sera for BTV antibody detection and RNA from red cells for BTV antigen detection.Erythrocyte preparations from BTV antigen-positive samples were inoculated into chicken embryo veins and then passaged three times in BHK-21 cells to isolate virus.Full-length c DNA copies of seg-2 from XJ1407were synthesized and sequenced as previously described(Maan,et al.,2007).Results:One hundred and twenty-six blood samples were collected from healthy sheep and goats in Xinjiang,China,during July of 2014.Seventy-three samples(57.93%)were BTV VP7 antibody-positive and 39 samples(30.95%)were BTV antigen-positive.BTV strain XJ1407 was isolated from the blood of BTV antigen-positive goats.Virus neutralization test(VNT)results showed that XJ1407 strain was not neutralized by BTV14 reactive antisera.Neutralization tests for XJ1407 using BTV serotype 257 positive sera were not assessed due to the lack of characterized antisera and control viruses in China.Sequence analysis of seg-2 of XJ1407 showed 75%nucleotide identity and 78%amino acid identity with BTV27 isolate obtained in 2014.Conclusions:The results of VNT and sequence analysis suggest XJ1407 may be a novel BTV serotype.

关键词： Bluetongue virus Epidemiology New serotype

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Establishment of an indirect immunofluorescence assay for the detection of African swine fever virus antibodies

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Journal of Integrative Agriculture 2024年第1期23卷 228-238页

作者： Wan Wang Zhenjiang Zhang Weldu Tesfagaber Jiwen Zhang Fang Li Encheng Sun Lijie Tang Zhigao Bu Yuanmao Zhu Dongming Zhao State Key Laboratory for Animal Disease Control and Prevention National African Swine Fever Para-reference LaboratoryNational High Containment Facilities for Animal Diseases Control and PreventionHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China College of Veterinary Medicine Northeast Agricultural UniversityHarbin 150069China

African swine fever(ASF)continues to cause enormous economic loss to the global pig industry.Since there is no safe and effective vaccine,accurate and timely diagnosis of ASF is essential to implement control measures.Indirect immunofluorescence assay(IFA)is a gold standard serological method recommended by the World Organization for Animal Health(WOAH).In this study,we used primary fetal kidney cells to establish a wild boar cell line(BK2258)that supported the efficient replication of ASF virus(ASFV)SD/DY-I/21 and showed visible cytopathic effect(CPE).Moreover,using BK2258,we established a sensitive and specific IFA for ASFV antibody detection.To standardize and evaluate the performance of this assay,we used serum samples from pigs infected with the low virulent genotype I SD/DY-I/21 and genotype II HLJ/HRB1/20,and immunized with the vaccine candidate HLJ/18-7GD,field samples,and negative serum samples.The IFA reacted with the ASFV-positive sera and displayed bright fluorescence foci.There was no non-specific green fluorescence due to cellular senescence or other cell damage-causing factors.Compared to a commercial indirect enzyme-linked immunosorbent assay(iELISA),ASFV antibodies were detected 1–4 days earlier using our IFA.The detection limits of the IFA and iELISA for the same ASFV-antibody positive serum samples were 1:25,600 and 1:6,400,respectively,indicating that the IFA is more sensitive than iELISA.The newly established IFA was highly specific and did not cross-react with sera positive for six other important porcine pathogens(i.e.,Classical swine fever virus(CSFV),Porcine reproductive and respiratory syndrome virus(PRRSV),Porcme circovirus type 2(PCV2),Pseudorabies virus(PRV),Foot-and-Mouth disease virus type O(FMDV/O),and Porcine epidemic diarrhea virus(PEDV)).This study thus provides a sensitive,specific,and reliable detection method that is suitable for the serological diagnosis of ASF.

关键词： African swine fever antibody IFA serological method

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中国布鲁氏菌病的防控

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中国科学：生命科学 2023年第12期53卷 1713-1721页

作者：刘文兴步志高中国农业科学院哈尔滨兽医研究所动物疫病防控全国重点实验室哈尔滨150069

布鲁氏菌病(简称布病)是由胞内寄生的布鲁氏菌引起的一种重要人兽共患传染病,呈世界性分布,中东和亚洲地区流行严重,中国也不例外.革兰氏阴性的布鲁氏菌入侵宿主后,定植于巨噬细胞,逃避宿主免疫系统,造成慢性感染;可感染人、多种家畜和... 详细信息

布鲁氏菌病(简称布病)是由胞内寄生的布鲁氏菌引起的一种重要人兽共患传染病,呈世界性分布,中东和亚洲地区流行严重,中国也不例外.革兰氏阴性的布鲁氏菌入侵宿主后,定植于巨噬细胞,逃避宿主免疫系统,造成慢性感染;可感染人、多种家畜和野生动物,引致相似的临床症状与病理损伤.布病不仅危害动物和人类健康,而且影响经济发展与社会稳定,甚至威胁到国家安全.加强布病的科学研究与综合防控,既有经济学和公共卫生学意义,也是国家安全战略的重要内容之一.传染病防控,既遵循“消灭传染源、保护易感宿主和切断传播途径”三个基本原则,又是一项系统工程;其中,良好的监测、可靠的诊断和安全有效的疫苗尤为重要.中国自1950年始,建立了科研队伍对布病进行调查与研究,开展联合攻关,分阶段、有步骤地采取综合防控措施,取得丰硕成果的同时,也积累出经验与教训;制定并更新了布鲁氏菌病诊断技术的行业和国家标准,相继有布鲁氏菌cELISA,iELISA和胶体金的抗体检测试剂盒(试纸条)获得新兽药证书;同时也是独立自主成功研制布鲁氏菌弱毒疫苗的国家,相继有布鲁氏菌常规弱毒疫苗(M5,S2和M5-90)和基因缺失活疫苗(A19-ΔVirB12,M5-90Δ26和BA0711)批准使用.这些研究成果的推广应用为布鲁氏菌有效控制和最终净化提供科技支撑.

关键词：布鲁氏菌病人兽共患疫苗诊断防控

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守正创新笃行致远——中国农业科学院哈尔滨兽医研究所

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中国科学：生命科学 2023年第12期53卷 1697-1699页

作者：柳金雄步志高中国农业科学院哈尔滨兽医研究所哈尔滨150069

1946年,为支援解放战争和战后恢复生产,原延安光华农场场长陈凌风同志受党中央委派,辗转来到东北解放区.经过一年多的艰苦筹建,于1948年在哈尔滨伪满家畜防疫所的废址上成立了“东北行政委员会农林处家畜防治所”并担任第一任所长.研究... 详细信息

1946年,为支援解放战争和战后恢复生产,原延安光华农场场长陈凌风同志受党中央委派,辗转来到东北解放区.经过一年多的艰苦筹建,于1948年在哈尔滨伪满家畜防疫所的废址上成立了“东北行政委员会农林处家畜防治所”并担任第一任所长.研究所历经多次更名——“东北人民政府农业部兽医研究所”(1949年)、“哈尔滨兽医研究所”(1954年,隶属中央政府农业部)、“中国农业科学院兽医研究所”(1957年,隶属中国农业科学院),于1965年定名为“中国农业科学院哈尔滨兽医研究所”(哈兽研).

关键词：中国农业科学院兽医研究所哈兽研东北解放区防治所守正创新

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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

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Mouse-adapted SARS-CoV-2 replicates efficiently in the upper and lower respiratory tract of BALB/c and C57BL/6J mice

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Protein & Cell 2020年第10期11卷 776-782页

作者： Jinliang Wang Lei Shuai Chong Wang Renqiang Liu Xijun He Xianfeng Zhang Ziruo Sun Dan Shan Jinying Ge Xijun Wang Ronghong Hua Gongxun Zhong Zhiyuan Wen Zhigao Bu State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences Harbin 150069 China National High Containment Laboratory for Animal Diseases Control and Prevention Harbin 150069 China

Dear Editor,As of June,2020,more than ten million cases of COVID-19 have been reported worldwide.The causative pathogen of the disease is a novel coronavirus named severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)(World Health Organization,2020).Animal infection models are important to characterize the infection,pathogenesis,and immunology of SARS・CoV・2,as well as for the development of medications and vaccines against COVID-19.Mice are particularly attractive animal models for their identical genetic background,reliable reproducibility,well characterized biology,and the huge availability of research reagents and knockout animals.Models in inbreed mice such as BALB/c and C57BL/6J(C57),which are widely used in research,are highly desired.

关键词： C57BL/6J BALB/c respiratory

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A seven-gene-deleted African swine fever virus is safe and effective as a live attenuated vaccine in pigs

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Science China(Life Sciences) 2020年第5期63卷 623-634页

作者： Weiye Chen Dongming Zhao Xijun He Renqiang Liu Zilong Wang Xianfeng Zhang Fang Li Dan Shan Hefeng Chen Jiwen Zhang Lulu Wang Zhiyuan Wen Xijun Wang Yuntao Guan Jinxiong Liu Zhigao Bu State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150001China National High Containment Laboratory for Animal Diseases Control and Prevention Harbin 150069China

African swine fever(ASF)is a devastating infectious disease in swine that is severely threatening the global pig industry.An efficacious vaccine is urgently required.Here,we used the Chinese ASFV HLJ/18 as a backbone and generated a series of genedeleted viruses.The virulence,immunogenicity,safety,and protective efficacy evaluation in specific-pathogen-free pigs,commercial pigs,and pregnant sows indicated that one virus,namely HLJ/18-7GD,which has seven genes deleted,is fully attenuated in pigs,cannot convert to the virulent strain,and provides complete protection of pigs against lethal ASFV challenge.Our study shows that HLJ/-18-7GD is a safe and effective vaccine against ASFV,and as such is expected to play an important role in controlling the spread of ASFV.

关键词： African swine fever vaccine safety protective efficacy pig

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