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Acta Pharmaceutica Sinica B 2017年第5期7卷 571-582,616页

作者： Congjie Zhai Ling Ma Zhixin Zhang Jiwei Ding Jing Wang Yongxin Zhang Xiaoyu Li Fei Guo Liyan Yu Jinming Zhou Shan Cen institute of Medicinal Biotechnology Chinese Academy of Medical Science Institute of Pathogen Biology Chinese Academy of Medical Science

Human APOBEC3G(h A3G) is a cytidine deaminase which inhibits HIV-1 replication. The HIV-1 accessory protein viral infectivity factor(Vif) counteracts with hA3G by targeting it for proteasomal degradation. In this work, we constructed and optimized molecular models of the hA3G dimer and the h A3G–Vif complex. The molecular modeling study revealed that the loop7 motif of hA3G appears on the interfaces of both the h A3G–Vif complex and the hA3G dimer. Biochemical analysis provided evidence suggesting that binding of Vif to hA3G results in steric blocking of hA3G dimerization, implying that monomeric hA3G serves as a substrate for Vif-mediated degradation.Furthermore, we presented evidence for the important roles of the loop7 motif, especially the central residues within the region, in hA3G dimerization, h A3G–Vif interaction, Vif-mediated hA3G degradation as well as subcellular localization of hA3G. This work highlights a multiple-task interface formed by loop7 motif, which regulates biological function of hA3G, thus providing the feasibility of the strategy of blocking Vif-mediated A3 G degradation by targeting the putative site around loop7.

关键词： HIV Vif APOBEC3G Motif Modeling hA3G–Vif complex hA3G dimer

Generation of HIY-resistant cells with a single-domain antibody:implications for HIV-1 gene therapy

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Cellular & Molecular Immunology 2021年第3期18卷 660-674页

作者： Hongliang Jin Xiaoran Tang Li Li Yue Chen Yuanmei Zhu Huihui Chong and Yuxian He NHC Key Laboratory of Systems biology of pathogen Institute of Pathogen Biology Chinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina Center for AIDS Research Chinese Academy of Medical Sciences and Peking Union Medical ColleyBeijingChina

The cure or functional cure of the"Berlin patient"and"London patient"indicates that infusion of HIV-resistant cells could be a viable treatment strategy.Very recently,we genetically linked a short-peptide fusion inhibitor with a glycosylphosphatidylinositol(GPI)attachment signal,rendering modified cells fully resistant to HIV infection.In this study,GPI-anchored m36.4,a single-domain antibody(nanobody)targeting the coreceptor-binding site of gp120,was constructed with a lentiviral vector.We verified that m36.4 was efficiently expressed on the plasma membrane of transduced TZM-bl cells and targeted lipid raft sites without affecting the expression of HIV receptors(CD4,CCR5,and CXCR4).Significantly,TZM-bl cells expressing GPI-m36.4 were highly resistant to infection with divergent HIV-1 subtypes and potently blocked HIV-1 envelope-mediated cell-cell fusion and cell-cell viral transmission.Furthermore,we showed that GPI-m36.4-modified human CEMss-CCR5 cells were nonpermissive to both CCR5-and CXCR4-tropic HIV-1 isolates and displayed a strong survival advantage over unmodified cells.It was found that GPI-m36.4 could also Impair HIV-1 Env processing and viral infectivity in transduced cells,underlying a multifaceted mechanism of antiviral action.In conclusion,our studies characterize m36.4 as a powerful nanobody that can generate HIV-resistant cells,offering a novel gene therapy approach that can be used alone or in combination.

关键词： HIV-1 gene therapy resistant cell single-domain antibody glycosylphosphatidylinositol

EV713C protease cleaves host anti-viral factor OAS3 and enhances virus replication

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Virologica Sinica 2022年第3期37卷 418-426页

作者： Xiaolei Zhou Li Tian Jian Wang Baisong Zheng Wenyan Zhang Center for Infectious Diseases and pathogen biology Institute of Virology and AIDS ResearchKey Laboratory of Organ Regeneration and Transplantation of the Ministry of EducationThe First Hospital of Jilin UniversityJilin130021China

The global spread of enteroviruses(EVs)has become more frequent,severe and life-threatening.Intereron(IFN)I has been proved to control EVs by regulating IFN-stimulated genes(ISG)expression.20-50-oligoadenylate synthetases 3(OAS3)is an important ISG in the OAS/RNase L antiviral system.The relationship between OAS3 and EVs is still unclear.Here,we reveal that OAS3,superior to OAS1 and OAS2,significantly inhibited EV71 replication in vitro.However,EV71 utilized autologous 3C protease(3C^(pro))to cleave intracellular OAS3 and enhance viral replication.Rupintrivir,a human rhinovirus 3C protease inhibitor,completely abolished the cleavage of EV713C^(pro)on OAS3.And the proteolytically deficient mutants H40G,E71A,and C147G of EV713C^(pro)also lost the ability of OAS3 cleavage.Mechanistically,the Q982-G983 motif in C-terminal of OAS3 was identified as a crucial 3C^(pro)cutting site.Further investigation indicated that OAS3 inhibited not only EV71 but also Coxsackievirus B3(CVB3),Coxsackievirus A16(CA16),Enterovirus D68(EVD68),and Coxsackievirus A6(CA6)subtypes.Notably,unlike other four subtypes,CA163C^(pro)could not cleave OAS3.Two key amino acids variation Ile36 and Val86 in CA163C^(pro)might result in weak and delayed virus replication of CA16 because of failure of OAS and 3AB cleavage.Our works elucidate the broad anti-EVs function of OAS3,and illuminate a novel mechanism by which EV71 use 3C^(pro)to escape the antiviral effect of OAS3.These findings can be an important entry point for developing novel therapeutic strategies for multiple EVs infection.

关键词： Enteroviruses EV71 3C protease Cleavage Anti-viral factor OAS3

IFN-β1b induces OAS3 to inhibit EV71 via IFN-β1b/JAK/STAT1 pathway

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Virologica Sinica 2022年第5期37卷 676-684页

作者： Baisong Zheng Xiaolei Zhou Li Tian Jian Wang Wenyan Zhang Center for pathogen biology and Infectious Diseases Institute of Virology and AIDS ResearchKey Laboratory of Organ Regeneration and Transplantation of the Ministry of EducationThe First Hospital of Jilin UniversityChangchun130021China

Enterovirus 71(EV71) caused hand, foot and mouth disease(HFMD) is a serious threat to the health of young children. Although type I interferon(IFN-I) has been proven to control EV71 replication, the key downstream IFNstimulated gene(ISG) remains to be clarified and investigated. Recently, we found that 2’-5’-oligoadenylate synthetases 3(OAS3), as one of ISG of IFN-β1b, was antagonized by EV71 3C protein. Here, we confirm that OAS3is the major determinant of IFN-β1b-mediated EV71 inhibition, which depends on the downstream constitutive RNase L activation. 2’-5’-oligoadenylate(2-5A) synthesis activity deficient mutations of OAS3 D816A, D818A,D888A, and K950A lost resistance to EV71 because they could not activate downstream RNase L. Further investigation proved that EV71 infection induced OAS3 but not RNase L expression by IFN pathway. Mechanically, EV71 or IFN-β1b-induced phosphorylation of STAT1, but not STAT3, initiated the transcription of OAS3 by directly binding to the OAS3 promoter. Our works elucidate the immune regulatory mechanism of the host OAS3/RNase L system against EV71 replication.

关键词： Enterovirus 71(EV71) 20-50-oligoadenylate synthetases 3(OAS3) RNase L IFN-β1b JAK/STAT

The taxonomy, host range and pathogenicity of coronaviruses and other viruses in the Nidovirales order

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Animal Diseases 2021年第1期1卷 3-30页

作者： Zhijian Zhou Ye Qiu Xingyi Ge Hunan Provincial Key Laboratory of Medical Virology Institute of Pathogen Biology and ImmunologyCollege of BiologyHunan University27 Tianma Rd.ChangshaHunanChina

The frequent emergence of coronavirus(CoV)epidemics has seriously threatened public health and stock farming.The major hosts for CoVs are birds and mammals.Although most CoVs inhabit their specific natural hosts,some may occasionally cross the host barrier to infect livestock and even people,causing a variety of diseases.Since the beginning of the new century,increasing attention has been given to research on CoVs due to the emergence of highly pathogenic and genetically diverse CoVs that have caused several epidemics,including the recent COVID-19 pandemic.CoVs belong to the Coronaviridae family of the Nidovirales order.Recently,advanced techniques for viral detection and viral genome analyses have enabled characterization of many new nidoviruses than ever and have greatly expanded the Nidovirales order with new classification and nomenclature.Here,we first provide an overview of the latest research progress in the classification of the Nidovirales order and then introduce the host range,genetic variation,genomic pattern and pathogenic features of epidemic CoVs and other epidemic viruses.This information will promote understanding of the phylogenetic relationship and infectious transmission of various pathogenic nidoviruses,including epidemic CoVs,which will benefit virological research and viral disease control.

关键词： Coronavirus Nidovirales Hosts S protein Genetics Evolution

Inhibition of dengue virus replication by diisopropyl chrysin-7-yl phosphate

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Science China(Life Sciences) 2016年第8期59卷 832-838页

作者： Jiang Du Zhe Chen Ting Zhang Iianmin Wang Qi Jin MOH Key Laboratory of Systems biology of pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100176China

Dengue fever is a tropical disease and caused by dengue virus(DENV), which is transmitted by mosquitoes and infects about 400 million people annually. With the development of international trade and travel, China is facing a growing threat. Over 40 thousands of people were infected during the 2014 DENV outbreak in Guangdong. Neither licensed vaccine nor therapeutic drug has been available. In this report, we isolated two clinical DENV strains. The full-length genome was sequenced and characterized. We also applied a flavonoid, CPI, into an anti-DENV assay. Replication of viral RNA and expression of viral protein was all strongly inhibited. These results indicated that CPI may serve as potential protective agents in the treatment of patients with chronic DENV infection.

关键词： dengue virus virus isolation phylogenetic analysis antiviral activity flavonoid

Host restriction factors for hepatitis C virus

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World Journal of Gastroenterology 2016年第4期22卷 1477-1486页

作者： li-ya zhou lei-liang zhang MOH Key Laboratory of Systems biology of pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100176China

Host-hepatitis C virus(HCV) interactions have both informed fundamental concepts of viral replication and pathogenesis and provided novel insights into host cell biology. These findings are illustrated by the recent discovery of host-encoded factors that restrict HCV infection. In this review, we briefly discuss these restriction factors in different steps of HCV infection. In each case, we discuss how these restriction factors were identified, the mechanisms by which they inhibit HCV infection and their potential contribution to viral pathogenesis.

关键词： Hepatitis C virus Host restriction factor Interferon Entry Replication Propagation

Armigeres subalbatus is a potential vector for Zika virus but not dengue virus

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Infectious Diseases of Poverty 2022年第3期11卷 92-92页

作者： Wenqiang Yang Siyu Zhao Yugu Xie Tong Liu Ling Kong Yijia Guo Zhensheng Xie Peiwen Liu Xiao-Guang Chen Department of pathogen biology Institute of Tropical MedicineSchool of Public HealthSouthern Medical UniversityGuangzhouChina

Background:Zika virus (ZIKV) and dengue virus (DENV) are closely related flaviviruses primarily transmitted by Aedes mosquitoes. Armigeres subalbatus is an emerging and widely distributed mosquito, and ZIKV has been detected and isolated from it. However, it is not clear whether Ar. subalbatus could be a vector for ZIKV and DENV or not. In this study, we investigated the infection and transmission of Ar. subalbatus to ZIKV and DENV.Methods:A line of Ar. subalbatus was isolated from Guangdong, China, and further identified by the mitochondrial cytochrome oxidase subunit 1 (COI) gene. The adults of Ar. subalbatus were fed with blood meal containing ZIKV or DENV-2. At 4, 7, 10, 14, and 21 days post-inoculation (dpi), the infections of ZIKV or DENV-2 in the midguts, ovaries and salivary glands were detected and quantified by RT-PCR and RT-qPCR. To assess the transmissibility, suckling mice were exposed to bites of ZIKV-infected mosquitoes, and ZIKV was detected in brain tissue by RT-qPCR and plaque assays. Furthermore, the larvae of Ar. subalbatus were reared in artificial urine containing ZIKV or DENV-2. The infection rates and viral titers of larvae and adults were analyzed by RT-PCR and RT-qPCR, and the viral distribution in larval tissues was observed by immunohistochemistry. Chi-square test and one-way ANOVA analysis were used for assessing the infection rate and viral titer in varied tissues and different time points, respectively.Results:Following oral inoculation, ZIKV but not DENV-2 could be detected in Ar. subalbatus midguts at 4 dpi, ovaries at 7 dpi and salivary glands at 10 dpi. The highest infection rate (IR) of ZIKV was 27.8% in midgut at 7 dpi, 9.7% in ovary and 5.6% in salivary gland at 21 dpi. Eight days after being bitten by ZIKV-positive mosquitoes, ZIKV was detected in three brain tissues out of four suckling mice exposed to bites. ZIKV could be detected in the larvae reared in artificial urine contained ZIKV at a high concentration of 105 pfu/ml and various tissues of adults with a low infection

关键词： Armigeres subalbatus Zika virus Dengue virus Vector

Application of Nanopore Sequencing Technology in the Clinical Diagnosis of Infectious Diseases

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Biomedical and Environmental Sciences 2022年第5期35卷 381-392页

作者： ZHANG Lu Lu ZHANG Chi PENG Jun Ping NHC Key Laboratory of Systems biology of pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100176China Key Laboratory of Respiratory Disease pathogenomics Chinese Academy of Medical SciencesBeijing 100176China

Infectious diseases are an enormous public health burden and a growing threat to human health worldwide.Emerging or classic recurrent pathogens,or pathogens with resistant traits,challenge our ability to diagnose and control infectious diseases.Nanopore sequencing technology has the potential to enhance our ability to diagnose,interrogate,and track infectious diseases due to the unrestricted read length and system portability.This review focuses on the application of nanopore sequencing technology in the clinical diagnosis of infectious diseases and includes the following:(i)a brief introduction to nanopore sequencing technology and Oxford Nanopore Technologies(ONT)sequencing platforms;(ii)strategies for nanopore-based sequencing technologies;and(iii)applications of nanopore sequencing technology in monitoring emerging pathogenic microorganisms,molecular detection of clinically relevant drug-resistance genes,and characterization of disease-related microbial communities.Finally,we discuss the current challenges,potential opportunities,and future outlook for applying nanopore sequencing technology in the diagnosis of infectious diseases.

关键词： Nanopore sequencing Infectious diseases pathogen Oxford Nanopore Technologies

Occurrence and distribution of geminiviruses in China

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Science China(Life Sciences) 2022年第8期65卷 1498-1503页

作者： Fangfang Li Rui Qiao Zhanqi Wang Xiuling Yang Xueping Zhou State Key Laboratory for biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China Key Laboratory of Vector biology and pathogen Control of Zhejiang Province College of Life SciencesHuzhou UniversityHuzhou 313000China State Key Laboratory of Rice biology Institute of BiotechnologyZhejiang UniversityHangzhou 310058China

Geminiviruses are a group of plant viruses that cause severe diseases in many economically important crops worldwide, leading to devastating losses to agricultural production. Here we summarize the occurrence and distribution of geminiviruses in China,which provides valuable information for further epidemiological studies and supports the development of effective disease management strategies.

关键词： geminiviruses occurrence distribution China