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Screening and identification of HTNV_(pv)entry inhibitors with high-throughput pseudovirus-based chemiluminescence

Virologica Sinica 2022年第4期37卷 531-537页

作者： Xiaojing Wen Li Zhang Qiang Liu Xinyue Xiao Weijin Huang youchun wang Division of HIV/AIDS and Sex-transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)and WHO Collaborating Center for Standardization and Evaluation of BiologicalsBeijing 102629China Department of Microbiological Laboratory Technology School of Public HealthShandong UniversityJinan 250012China Institute for Reference Standards and Standardization National Institutes for Food and Drug ControlBeijing 102629China

Hantaviruses,such as Hantaan virus(HTNV)and Seoul virus,are the causative agents of Hantavirus cardiopulmonary syndrome(HCPS)and hemorrhagic fever with renal syndrome(HFRS),and are important zoonotic pathogens.China has the highest incidence of HFRS,which is mainly caused by HTNV and Seoul virus.No approved antiviral drugs are available for these hantaviral diseases.Here,a chemiluminescence-based highthroughput-screening(HTS)assay was developed and used to screen HTNV pseudovirus(HTNVpv)inhibitors in a library of 1813 approved drugs and 556 small-molecule compounds from traditional Chinese medicine sources.We identified six compounds with in vitro anti-HTNVpvactivities in the low-micromolar range(EC50values of0.1–2.2μmol/L;selectivity index of 40–900).Among the six selected compounds,cepharanthine not only showed good anti-HTNVpvactivity in vitro but also inhibited HTNVpv-fluc infection in Balb/c mice 5 h after infection by94%(180 mg/kg/d,P<0.01),93%(90 mg/kg/d,P<0.01),or 92%(45 mg/kg/d,P<0.01),respectively,in a bioluminescent imaging mouse model.A time-of-addition analysis suggested that the antiviral mechanism of cepharanthine involves the membrane fusion and entry phases.Overall,we have established a HTS method for antiviral drugs screening,and shown that cepharanthine is a candidate for HCPS and HFRS therapy.These findings may offer a starting point for the treatment of patients infected with hantaviruses.

关键词： Hantaan virus(HTNV) High-throughput screening(HTS) Inhibitor Drug repurposing

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A fresh class of superconducting and hard pentaborides

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Matter and Radiation at Extremes 2023年第5期8卷 89-95页

作者： Hui Xie Hong wang Fang Qin Wei Han Suxin wang youchun wang Fubo Tian Defang Duan College of Physics and Electronic Engineering Hebei Normal University for NationalitiesChengde 067000China College of Physics and Electronic Engineering Linyi UniversityLinyi 276000China College of Physics Jilin UniversityChangchun 130012China

On the basis of the current theoretical understanding of boron-based hard superconductors under ambient conditions,numerous studies have been conducted with the aim of developing superconducting materials with favorable mechanical properties using boron-rich compounds.In this paper,first-principles calculations reveal the existence of an unprecedented family of tetragonal pentaborides MB_(5)(M=Na,K,Rb,Ca,Sr,Ba,Sc,and Y),comprising B_(20)cages and centered metal atoms acting as stabilizers and electron donors to the boron sublattice.These compounds exhibit both superconductivity and high hardness,with the maximum superconducting transition temperature T_(c)of 18.6 K being achieved in RbB5 and the peak Vickers hardness Hv of 35.1 GPa being achieved in KB_(5)at 1 atm.The combination of these properties is particularly evident in KB_(5),RbB5,and BaB5,with Tc values of∼14.7,18.6,and 16.3 K and H_(v)values of∼35.1,32.4,and 33.8 GPa,respectively.The results presented here reveal that pentaborides can provide a framework for exploring and designing novel superconducting materials with favorable hardness at achievable pressures and even under ambient conditions.

关键词： boride boron superconducting

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Animal models for COVID-19:advances,gaps and perspectives

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Signal Transduction and Targeted Therapy 2022年第8期7卷 2817-2840页

作者： Changfa Fan Yong Wu Xiong Rui Yuansong Yang Chen Ling Susu Liu Shunan Liu youchun wang Division of Animal Model Research Institute for Laboratory Animal ResourcesNational Institutes for Food and Drug Control(NIFDC)National Rodent Laboratory Animal Resources CenterBeijing 102629China Department of Microbiology&Infectious Disease Center School of Basic Medical SciencesPeking University Health Science CenterBeijing 100083China College of Life Sciences Northwest UniversityProvincial Key Laboratory of Biotechnology of Shaanxi ProvinceNorthwest UniversityXi’an 710069China Division of HIV/AIDS and Sexually Transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)BeijingChina

COVID-19,caused by SARS-CoV-2,is the most consequential pandemic of this century.Since the outbreak in late 2019,animal models have been playing crucial roles in aiding the rapid development of vaccines/drugs for prevention and therapy,as well as understanding the pathogenesis of SARS-CoV-2 infection and immune responses of hosts.However,the current animal models have some deficits and there is an urgent need for novel models to evaluate the virulence of variants of concerns(VOC),antibodydependent enhancement(ADE),and various comorbidities of COVID-19.This review summarizes the clinical features of COVID-19 in different populations,and the characteristics of the major animal models of SARS-CoV-2,including those naturally susceptible animals,such as non-human primates,Syrian hamster,ferret,minks,poultry,livestock,and mouse models sensitized by genetically modified,AAV/adenoviral transduced,mouse-adapted strain of SARS-CoV-2,and by engraftment of human tissues or cells.Since understanding the host receptors and proteases is essential for designing advanced genetically modified animal models,successful studies on receptors and proteases are also reviewed.Several improved alternatives for future mouse models are proposed,including the reselection of alternative receptor genes or multiple gene combinations,the use of transgenic or knock-in method,and different strains for establishing the next generation of genetically modified mice.

关键词： prevention drugs establishing

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Monkeypox virus quadrivalent mRNA vaccine induces immune response and protects against vaccinia virus

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Signal Transduction and Targeted Therapy 2023年第5期8卷 2423-2433页

作者： Ye Sang Zhen Zhang Fan Liu Haitao Lu Changxiao Yu Huisheng Sun Jinrong Long Yiming Cao Jierui Mai Yiqi Miao Xin wang Jiaxin Fang youchun wang Weijin Huang Jing Yang Shengqi wang Bioinformatics center of AMMS Beijing100850P.R.China Division of HIV/AIDS and Sex-transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)Beijing102629P.R.China Institute of Medical Biology Chinese Academy of Medical SciencesKunming650031P.R.China

Monkeypox has been declared a public health emergency by the World Health Organization.There is an urgent need for efficient and safe vaccines against the monkeypox virus(MPXV)in response to the rapidly spreading monkeypox epidemic.In the age of COVID-19,mRNA vaccines have been highly successful and emerged as platforms enabling rapid development and large-scale preparation.Here,we develop two MPXV quadrivalent mRNA vaccines,named mRNA-A-LNP and mRNA-B-LNP,based on two intracellular mature virus specific proteins(A29L and M1R)and two extracellular enveloped virus specific proteins(A35R and B6R).By administering mRNA-A-LNP and mRNA-B-LNP intramuscularly twice,mice induce MPXV specific IgG antibodies and potent vaccinia virus(VACV)specific neutralizing antibodies.Further,it elicits efficient MPXV specific Th-1 biased cellular immunity,as well as durable effector memory T and germinal center B cell responses in mice.In addition,two doses of mRNA-A-LNP and mRNA-B-LNP are protective against the VACV challenge in mice.And,the passive transfer of sera from mRNA-A-LNP and mRNA-B-LNP-immunized mice protects nude mice against the VACV challenge.Overall,our results demonstrate that mRNA-A-LNP and mRNA-B-LNP appear to be safe and effective vaccine candidates against monkeypox epidemics,as well as against outbreaks caused by other orthopoxviruses,including the smallpox virus.

关键词： doses protective immunity

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Cross-reactivity of eight SARS-CoV-2 variants rationally predicts immunogenicity clustering in sarbecoviruses

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Signal Transduction and Targeted Therapy 2022年第8期7卷 3069-3079页

作者： Qianqian Li Li Zhang Ziteng Liang Nan wang Shuo Liu Tao Li Yuanling Yu Qianqian Cui Xi Wu Jianhui Nie Jiajing Wu Zhimin Cui Qiong Lu Xiangxi wang Weijin Huang youchun wang Division of HIV/AIDS and Sex-transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)WHO Collaborating Center for Standardization and Evaluation of BiologicalsNHC Key Laboratory of Research on Quality and Standardization of Biotech Products and NMPA Key Laboratory for Quality Research and Evaluation of Biological Products102629 BeijingChina Jiangsu Recbio Technology Co. Ltd.215300 TaizhouChina Graduate School of Chinese Academy of Medical Sciences&Peking Union Medical College No.9 Dongdan SantiaoDongcheng District100730 BeijingChina CAS Key Laboratory of Infection and Immunity CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of Sciences100101 BeijingChina

A steep rise in Omicron reinfection cases suggests that this variant has increased immune evasion ability.To evaluate its antigenicity relationship with other variants,antisera from guinea pigs immunized with spike protein of SARS-CoV-2 variants of concern(VOCs)and variants of interest(VOIs)were cross-tested against pseudotyped variants.The neutralization activity against Omicron was markedly reduced when other VOCs or VOIs were used as immunogens,and Omicron(BA.1)-elicited sera did not efficiently neutralize the other variants.However,a Beta or Omicron booster,when administered as the 4th dose 3-months after the 3rd dose of any of the variants,could elicit broad neutralizing antibodies against all of the current variants including Omicron BA.1.Further analysis with 280 available antigen–antibody structures and quantification of immune escape from 715 reported neutralizing antibodies provide explanations for the observed differential immunogenicity.Three distinct clades predicted using an in silico algorithm for clustering of sarbecoviruses based on immune escape provide key information for rational design of vaccines.

关键词： rational escape neutral

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The antigenicity of SARS-CoV-2 Delta variants aggregated 10 high-frequency mutations in RBD has not changed sufficiently to replace the current vaccine strain

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Signal Transduction and Targeted Therapy 2022年第2期7卷 492-501页

作者： Jiajing Wu Jianhui Nie Li Zhang Hao Song Yimeng An Ziteng Liang Jing Yang Ruxia Ding Shuo Liu Qianqian Li Tao Li Zhimin Cui Mengyi Zhang Peng He youchun wang Xiaowang Qu Zhongyu Hu Qihui wang Weijin Huang Division of HIV/AIDS and Sex-transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)102629 BeijingChina Research Network of Immunity and Health(RNIH) Beijing Institutes of Life ScienceChinese Academy of Sciences100101 BeijingChina CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of MicrobiologyCenter for Influenza Research and Early-warning(CASCIRE)CAS-TWAS Center of Excellence for Emerging Infectious Diseases(CEEID)Chinese Academy of Sciences100101 BeijingChina Division of Hepatitis and Enteric Viral Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)and Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products102629 BeijingChina Translational Medicine Institute First People’s Hospital of ChenzhouUniversity of South ChinaChenzhouChina CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of MicrobiologyChinese Academy of Sciences100101 BeijingChina

Emerging SARS-CoV-2 variants are the most serious problem for COVID-19 prophylaxis and treatment.To determine whether the SARS-CoV-2 vaccine strain should be updated following variant emergence like seasonal flu vaccine,the changed degree on antigenicity of SARS-CoV-2 variants and H3N2 flu vaccine strains was compared.The neutralization activities of Alpha,Beta and Gamma variants’spike protein-immunized sera were analysed against the eight current epidemic variants and 20 possible variants combining the top 10 prevalent RBD mutations based on the Delta variant,which were constructed using pseudotyped viruses.Meanwhile,the neutralization activities of convalescent sera and current inactivated and recombinant protein vaccine-elicited sera were also examined against all possible Delta variants.Eight HA protein-expressing DNAs elicited-animal sera were also tested against eight pseudotyped viruses of H3N2 flu vaccine strains from 2011–2019.Our results indicate that the antigenicity changes of possible Delta variants were mostly within four folds,whereas the antigenicity changes among different H3N2 vaccine strains were approximately 10–100-fold.Structural analysis of the antigenic characterization of the SARS-CoV-2 and H3N2 mutations supports the neutralization results.This study indicates that the antigenicity changes of the current SARS-CoV-2 may not be sufficient to require replacement of the current vaccine strain.

关键词： vaccine antigenicity variants

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全球疫苗研发现状和展望

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中国药科大学学报 2024年第1期55卷 115-126页

作者：马磊杨昭庆王佑春中国医学科学院医学生物学研究所昆明650118

频繁暴发的新发突发传染病已成为影响人类公共卫生及健康的重要危险因素,目前接种疫苗依然是应对传染病最关键的手段。新型冠状病毒感染(COVID-19)疫情以来,无论是创新型疫苗基础技术还是疫苗研发策略均取得了革命性进展,以m RNA技术为... 详细信息

频繁暴发的新发突发传染病已成为影响人类公共卫生及健康的重要危险因素,目前接种疫苗依然是应对传染病最关键的手段。新型冠状病毒感染(COVID-19)疫情以来,无论是创新型疫苗基础技术还是疫苗研发策略均取得了革命性进展,以m RNA技术为代表的第三代疫苗技术逐渐成为一种新兴的疫苗研发方式。本文对近年来的不同疫苗技术特点进行全面梳理,综述了基于不同技术平台的重点疫苗产品的最新研究进展,并展望了未来疫苗研发策略,为疫苗研发工作提供经验和借鉴。

关键词：疫苗研发疫苗技术平台研究策略重点疫苗产品

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Structural and functional insights into MCR-2 mediated colistin resistance

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Science China(Life Sciences) 2018年第11期61卷 1432-1436页

作者： Xudong wang Qiong Lu Jianxun Qi Yan Chai youchun wang George Fu Gao CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of Microbiology Chinese Academy of Sciences College of Life Sciences University of Chinese Academy of Sciences Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products Division of HIV/AIDS and Sexually Transmitted Virus Vaccines National Institutes for Food and Drug Control National Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention (China CDC) Research Network of Immunity and Health (RNIH) Beijing Institutes of Life Science Chinese Academy of Sciences

Dear Editor,The spread of antibiotic resistance genes among bacteria is a serious and growing threat to global health (Peleg and Hoopet,2010).One emerging case is transferable genes found in bacteria-encoding enzymes resistant to colistin.Colistin is a cationic polypeptide antibiotic with broadspectrum activity against Gram-negative bacteria through interactions with the lipid A moiety of bacterial lipopoly-saccharide(LPS),subsequently destroying the cell envelope.During late 2015,a plasmid-mediated gene,mcr-1,was first reported from Escherichia coli(E.coli)in China to confer resistance to colistin(Liu et a1.,2016),and detected world-wide soon afterwards(Gao et a1.,2016;Hu et a1.,2016).

关键词： colistin resistance genes bacteria

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Immunological and metabolic characteristics of the Omicron variants infection

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Signal Transduction and Targeted Therapy 2023年第2期8卷 784-794页

作者： Jiejie Geng Xu Yang Kun wang Ke wang Ruo Chen Zhi-Nan Chen Chuan Qin Guizhen Wu youchun wang Ke Xu Peng Du Jiangning Liu Shirui Chen Tao Zhang Xiuxuan Sun Ting Guo Ying Shi Zheng Zhang Ding Wei Peng Lin Qingyi wang Jing Yuan Jiuxin Qu Jin Zou Yingxia Liu Hongzhou Lu Ping Zhu Huijie Bian Liang Chen Department of Cell Biology of National Translational Science Center for Molecular Medicine and Department of Clinical Immunology of Xijing Hospital Fourth Military Medical UniversityXi’an 710032China Key Laboratory of Human Disease Comparative Medicine Chinese Ministry of HealthKey Laboratory for Animal Models of Emerging and Remerging Infectious DiseasesInstitute of Laboratory Animal ScienceChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100871China MHC Key Laboratory of Biosafety National Institute for Viral Disease Control and Preven-tionChinese Center for Disease Control and PreventionBeijing 100871China Division of HIV/AIDS and Sex-transmitted Virus Vaccines Institute for Biological Product ControlNational Institutes for Food and Drug Control(NIFDC)and WHO Collaborating Center for Standardization and Evaluation of BiologicalsBeijing 102629China Beijing Institute of Biotechnology Beijing 100871China School of Basic Medicine Fourth Military Medical UniversityXi’an 710032China The Third People’s Hospital of Shenzhen Shenzhen 518112China School of Medicine Shanghai UniversityShanghai 200444China

The Omicron variants of SARS-CoV-2,primarily authenticated in November 2021 in South Africa,has initiated the 5th wave of global pandemics.Here,we systemically examined immunological and metabolic characteristics of Omicron variants infection.We found Omicron resisted to neutralizing antibody targeting receptor binding domain(RBD)of wildtype SARS-CoV-2.Omicron could hardly be neutralized by sera of Corona Virus Disease 2019(COVID-19)convalescents infected with the Delta variant.Through mass spectrometry on MHC-bound peptidomes,we found that the spike protein of the Omicron variants could generate additional CD8+T cell epitopes,compared with Delta.These epitopes could induce robust CD8+T cell responses.Moreover,we found booster vaccination increased the cross-memory CD8+T cell responses against Omicron.Metabolic regulome analysis of Omicron-specific T cell showed a metabolic profile that promoted the response of memory T cells.Consistently,a greater fraction of memory CD8+T cells existed in Omicron stimulated peripheral blood mononuclear cells(PBMCs).In addition,CD147 was also a receptor for the Omicron variants,and CD147 antibody inhibited infection of Omicron.CD147-mediated Omicron infection in a human CD147 transgenic mouse model induced exudative alveolar pneumonia.Taken together,our data suggested that vaccination booster and receptor blocking antibody are two effective strategies against Omicron.

关键词： infection inhibited neutral

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Three amino acid residues in the envelope of human immunodeficiency virus type 1 CRF07_BC regulate viral neutralization susceptibility to the human monoclonal neutralizing antibody IgG1b12

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Virologica Sinica 2014年第5期29卷 299-307页

作者： Jianhui Nie Juan Zhao Qingqing Chen Weijin Huang youchun wang College of Life Science Jilin University Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products Division of HIV/AIDS and sex-transmitted virus vaccines National Institutes for Food and Drug Control (NIFDC)

The CD4 binding site(CD4bs) of envelope glycoprotein(Env) is an important conserved target for anti-human immunodeficiency virus type 1(HIV-1) neutralizing antibodies. Neutralizing monoclonal antibodies IgG1 b12(b12) could recognize conformational epitopes that overlap the CD4 bs of Env. Different virus strains, even derived from the same individual, showed distinct neutralization susceptibility to b12. We examined the key amino acid residues affecting b12 neutralization susceptibility using single genome amplification and pseudovirus neutralization assay. Eleven amino acid residues were identified that affect the sensitivity of Env to b12. Through site-directed mutagenesis, an amino acid substitution at position 182 in the V2 region of Env was confirmed to play a key role in regulating the b12 neutralization susceptibility. The introduction of V182 L to a resistant strain enhanced its sensitivity to b12 more than twofold. Correspondingly, the introduction of L182 V to a sensitive strain reduced its sensitivity to b12 more than tenfold. Amino acid substitution at positions 267 and 346 could both enhance the sensitivity to b12 more than twofold. However, no additive effect was observed when the three site mutageneses were introduced into the same strain, and the sensitivity was equivalent to the single V182 L mutation. CRF07_BC is a major circulating recombinant form of HIV-1 prevalent in China. Our data may provide important information for understanding the molecular mechanism regulating the neutralization susceptibility of CRF07_BC viruses to b12 and may be helpful for a vaccine design targeting the CD4 bs epitopes.

关键词： human immunodeficiency virus type 1 CRF07_BC envelope glycoprotein IgG1b12 neutralizing antibody single genome amplification

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