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Heterologous interactions between NS1 proteins from different influenza a virus subtypes/strains

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Science China(Life Sciences) 2012年第6期55卷 507-515页

作者： LI WeiZhong ZHANG Heng WANG GeFei ZHANG Chi ZENG XiangXing LIU Hui CHEN XiaoXuan XU YanXuan LI KangSheng Department of Microbiology and Immunology Key Immunopathology Laboratory of Guangdong ProvinceShantou University Medical CollegeShantou 515041China Department of Veterinary Medicine University of MarylandCollege ParkMD 20742USA Department of Ultrasonic Central Hospital of Luoyang CityLuoyang 471009China

Non-structural protein 1 （NS1） of the influenza virus plays a crucial role in modulating the host immune response and facili- tating virus replication. The formation of a homodimer or an oligomer is necessary for NSI to exert its function efficiently. In the present study, the NS 1 protein from the A/Shantou/602/06（H3N2） virus （herein abbreviated as NS32） was found to interact with NS1 from A/Shantou/169/O6（H1N1）, A/Chicken/Guangdong/1/05（HSN1） and A/Quail/Hong Kong/G1/97（H9N2）（abbre- viated as NS11, NS51 and NS92, respectively） viruses, although NS32 shares 17.4%-20.9% sequence diversity with NS11, NS51 and NS92. This indicates that the heterologous interactions between NS1 proteins from different influenza a virus sub- types/strains may be a common event during co-infection.

关键词： influenza a virus NS1 protein heterologous interaction

A cell-based high-throughput approach to identify inhibitors of influenza a virus

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Acta Pharmaceutica Sinica B 2014年第4期4卷 301-306页

作者： Qian Gao Zhen Wang Zhenlong Liu Xiaoyu Li Yongxin Zhang Zhizhen Zhang Shan Cen Institute of Medicinal Biotechnology Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijingChina Department of Biochemistry Guangdong Medical CollegeZhanjiang524023China

influenza is one of the most common infections threatening public health worldwide and is caused by the influenza *** emergence of drug resistance has led to an urgent need to develop new anti-influenza *** this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA,which expresses Gaussia luciferase upon influenza a virus *** this cell line,an assay was developed and optimized to search for inhibitors of influenza virus *** studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format(Z 0 factor value40.8).A pilot screening provides further evidence for validation of the *** together,this work provides a simple,convenient,and reliable HTS assay to identify compounds with anti-influenza activity.

关键词： Cell-based assay Gaussia luciferase influenza a virus High-throughput screen

Cytokine Profiling in influenza a virus and Staphylococcal(Co-)Infections

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Infectious Microbes & Diseases 2022年第4期4卷 161-167页

作者： Lea A.Tölken Antje D.Paulikat Fabian Cuypers Sebastian B.Skorka Sven Hammerschmidt Nikolai Siemens Department of Molecular Genetics and Infection Biology Interfaculty Institute for Genetics and Functional GenomicsCenter for Functional Genomics of MicrobesUniversity of GreifswaldGreifswaldGermany

influenza a virus and Staphylococcus aureus are common causative agents of ***-infections with these two pathogens frequently occur and are characterized,among others,by higher morbidity and mortality due to hyper-inflammation of the ***,we aimed to profile systemic and local cytokine composition at early acute stages of pneumonia in *** recovered from single influenza a virus and/or staphylococcal *** contrast,co-infections led to a severe clinical *** distinct cytokine patterns were detected in lungs of single-pathogen-infected animals,co-infections combined both virus-and bacteria-driven ***,analyses of infected human primarymonocytic cells as well as bronchial epithelial cells did not reflectmurine *** on infectious dose,mainly bacteria-driven responses were *** impact of single cells to cytokine composition of the lungs and translation of murine studies to humans remains uncertain and warrants further studies.

关键词： influenza a virus Staphylococcus aureus co-infection cytokines

Structure and Function of the NS1 Protein of influenza a virus

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 155-162页

作者： Dongzi LIN~1 Jingfang LAN~2 Zhizhen ZHANG~1* 1 Department of Biochemistry and Molecular Biology,Guangdong Medical College,Zhanjiang 524023,China 2 Department of Clinical Laboratory,Hejin People's Hospital,Hejin 043300,China DepartmentofBiochemistryandMolecularBiology GuangdongMedicalCollegeZhanjiang524023China DepartmentofClinicalLaboratory HejinPeople'sHospitalHejin043300China

The avian influenza a virus currently prevailing in Asia causes fatal pneumonia and multipleorgan failure in birds and *** intensive research,understanding of the characteristics of influenzaA virus that determine its virulence is ***1A protein,a non-structural protein of influenza Avirus,was reported to contribute to its pathogenicity and ***1A protein is a multifunctionalprotein that plays a significant role in resisting the host antiviral response during the influenza *** briefly outlines the current knowledge on the structure and function of the NS1A protein.

关键词： influenza a virus NS1A protein RNA-binding domain effector domain dimer resistance antiviral response

Soluble interleukin-6 receptor is elevated during influenza a virus infection and mediates the IL-6 and IL-32 inflammatory cytokine burst

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Cellular & Molecular Immunology 2015年第5期12卷 633-644页

作者： Jun Wang Qing Wang Tao Han Yong-Kui Li Sheng-Li Zhu Fang Ao Jian Feng Ming-Zhen Jing Li Wang Lin-Bai Ye Ying Zhu The State Key Laboratory of Virology College of Life Sciences Wuhan University Wuhan China

influenza a virus （IAV） infection is a major worldwide public health problem. However, the factors involved in mediating the inflammatory response to this infection and their relationships remain poorly understood. Here, we show that IAV infection stimulates the expression of the soluble IL-6 receptor （slL-6R）, a multifunctional protein involved in IL-6 signaling. Interestingly, slL-6R expression upregulated the levels of its own ligand, IL-6 and those of the pro-inflammatory cytokine IL-32. shRNA-mediated knockdown of slL-6R suppressed I L-6 and IL-32, indicating that this regulation is dependent on slL-6R during IAV infection. Furthermore, our results demonstrate that IL-32 participates in a negative feedback loop that inhibits slL-6R while upregulating IL-6 expression during IAV infection. Therefore, we show that slL-6R is a critical cellular factor involved in the acute inflammatory response to viral infection.

关键词： IL-32 IL-6 inflammation network influenza a virus soluble IL-6 receptor

Evaluation of A Single-reaction Method for Whole Genome Sequencing of influenza a virus using Next Generation Sequencing

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Biomedical and Environmental Sciences 2016年第1期29卷 41-46页

作者： Zou Xiao Hui Chen Wen Bing Zhao Xiang Zhu Wen Fei Yang Lei Wang Da Yan Shu Yue Long chinese national influenza center national institute for viral disease control and preventionchina cdckey laboratory for medical virologyministry of healthBeijing 102206China

Objective To evaluate a single-reaction genome amplification method, the multisegment reverse transcription-PCR （M-RTPCR）, for its sensitivity to full genome sequencing of influenza a virus, and the ability to differentiate mix-subtype virus, using the next generation sequencing （NGS） platform. Methods virus genome copy was quantified and seria（iy diluted to different titers, followed by amplification with the M-RTPCR method and sequencing on the NGS platform. Furthermore, we manually mixed two subtype viruses to different titer rate and amplified the mixed virus with the M-RTPCR protocol, followed by whole genome sequencing on the NGS platform. We also used clinical samples to test the method performance. Results The M-RTPCR method obtained complete genome of testing virus at 125 copies/reaction and determined the virus subtype at titer of 25 copies/reaction. Moreover, the two subtypes in the mixed virus could be discriminated, even though these two virus copies differed by 200-fold using this amplification protocol. The sensitivity of this protocol we detected using virus RNA was also confirmed with clinical samples containing Iow-titer virus. Conclusion The M-RTPCR is a robust and sensitive amplification method for whole genome sequencing of influenza a virus using NGS platform.

关键词： influenza a virus Whole genome sequencing NGS

Down-regulation of miR-155 inhibits inflammatory response in human pulmonary microvascular endothelial cells infected with influenza a virus by targeting sphingosine-1-phosphate receptor 1

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Chinese Medical Journal 2020年第20期133卷 2429-2436页

作者： Si-Mei Shen Hao Jiang Jiang-Nan Zhao Yi Shi Department of Respiratory and Critical Care Medicine Jinling HospitalMedical School of Nanjing UniversityNanjingJiangsu 210016China Department of Emergency Medicine The Second Affiliated HospitalSoutheast UniversityNanjingJiangsu 210003China

Background:Endothelial cells play a key role in the cytokine storm caused by influenza a virus. MicroRNA-155 (miR-155) is an important regulator in inflammation. Its role in the inflammatory response to influenza A infection, however, has yet to be elucidated. In this study, we explored the role as well as the underlying mechanism of miR-155 in the cytokine production in influenza A-infected endothelial ***:Human pulmonary microvascular endothelial cells (HPMECs) were infected with the influenza a virus strain H1N1. The efficiency of H1N1 infection was confirmed by immunofluorescence. The expression levels of proinflammatory cytokines and miR-155 were determined using real-time polymerase chain reaction. A dual-luciferase reporter assay characterized the interaction between miR-155 and sphingosine-1-phosphate receptor 1 (S1PR1). Changes in the target protein levels were determined using Western blot ***:MiR-155 was elevated in response to the H1N1 infection in HPMECs (24 h post-infection vs. 0 h post-infection, 3.875 ± 0.062 vs. 1.043 ± 0.013, P = 0.001). Over-expression of miR-155 enhanced inflammatory cytokine production (miR-155 mimic vs. negative control, all P influenza A-induced inflammatory response by targeting S1PR1.

关键词： MicroRNA-155 Sphingosine 1-phosphate receptor 1 influenza a virus Endothelial cells

Mutation trend of hemagglutinin of influenza a virus: a review from a computational mutation viewpoint

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Acta Pharmacologica Sinica 2006年第5期27卷 513-526页

作者： Guang WU~1 Shao-min YAN Computational Mutation Project.DreamSciTech Consulting,301.Building 12.Nanyou A-zone,Jiannan Road.Shenzhen 518054,China Computational Mutation Project DreamSciTech Consulting 301 Building 12 Nanyou A-zone Jiannan Road Shenzhen 518054 China. hongguanglishibahao@***

Since 1999 we have developed two computational mutation approaches to ana-lyze the protein primary structure whose methodology and implications were re-viewed in *** first approach is the calculation of predictable and unpredict-able portions of amino-acid pairs in a protein,and the second is the calculation ofamino-acid distribution rank in a *** approaches provide quantitativemeasures to present a protein,which we have used to study a number of proteinswith numerous mutations such as p53 *** recently,we focussed ourefforts on analyzing the proteins mutating frequently over time such as hemagglu-tinins of influenza A *** this review we summarise our findings and theirimplications for hemagglutinin mutations in combination with some newly avail-able *** approaches throw light on the true nature of genetic heterogeneityof influenza virus hemagglutinins;that is,the protein variability is highly relevantto its amino-acid *** these approaches,we can monitor newmutations from influenza virus hemagglutinins and may predict their mutations inthe future.

关键词： amino acid sequence hemagglutinins influenza a virus mutation probability

Apoptosis in Raji cell line induced by influenza a virus

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Chinese Medical Journal 2003年第9期116卷 1321-1324页

作者：李虹肖丽英李华林李婉宜蒋中华张林李明远 Department of Microbiology West China Medical Center Sichuan University Chengdu 610041China Department of Forensic Medicine West China Medical CenterSichuan University Chengdu 610041China

OBJECTIVE: To study the apoptotic effects of influenza a virus on the Raji cell line. METHODS: Cultured Raji cells were infected with influenza a virus at a multiplicity of infection (m.o.i) of 20 and the effects of apoptosis were detected at different time points post infection using the following methods: electron microscope, DNA agarose gel electrophoresis, PI stained flow cytometry (FCM) and Annexin-V FITC/PI stained FCM. RESULTS: Raji cells infected with influenza a virus showed changes of morphology apoptosis, DNA agarose electrophoresis also demonstrated a ladder-like pattern of DNA fragments in a time-dependent manner. PI stained FCM showed 'apoptosis peak' and FITC/PI stained FCM showed apoptotic cells. Quantitative analysis indicated that the percentage of apoptotic Raji cells increased after infection, and cycloheximide (CHX), an eukaryotic transcription inhibitor, could effectively inhibit the apoptotic effects of influenza a virus in vitro. CONCLUSIONS: influenza a virus can induce apoptosis in Raji cell line suggesting that it may lead to a potential method for tumor therapy.

关键词： Apoptosis Humans influenza a virus Research Support, Non-U.S. Gov't Tumor Cells, Cultured