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M6PR interacts with the HA2 subunit of influenza a virus to facilitate the fusion of viral and endosomal membranes

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Science China(Life Sciences) 2024年第3期67卷 579-595页

作者： Yuzhen Hu Li Jiang Guangwen Wang Yangming Song Zhibo Shan Xuyuan Wang Guohua Deng Jianzhong Shi Guobin Tian Xianying Zeng Liling Liu Hualan Chen Chengjun Li State Key Laboratory for Animal Disease Control and Prevention Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China

influenza a virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membranes. In this study, we identified cation-dependent mannose-6-phosphate receptor(M6PR) as a crucial host factor for the replication of IAV. We found that siRNA knockdown of M6PR expression significantly reduced the growth titers of different subtypes of IAV, and that the inhibitory effect of M6PR siRNA treatment on IAV growth was overcome by the complement of exogenously expressed M6PR. When A549 cells were treated with siRNA targeting M6PR,the nuclear accumulation of viral nucleoprotein(NP) was dramatically inhibited at early timepoints post-infection, indicating that M6PR engages in the early stage of the IAV replication cycle. By investigating the role of M6PR in the individual entry and post-entry steps of IAV replication, we found that the downregulation of M6PR expression had no effect on attachment, internalization, early endosome trafficking,or late endosome acidification. However, we found that M6PR expression was critical for the fusion of viral envelope and late endosomal membranes. Of note, M6PR interacted with the hemagglutinin(HA) protein of IAV, and further studies showed that the lumenal domain of M6PR and the ectodomain of HA2 mediated the interaction and directly promoted the fusion of the viral and late endosomal membranes,thereby facilitating IAV replication. Together, our findings highlight the importance of the M6PR–HA interaction in the fusion of viral and late endosomal membranes during IAV replication.

关键词： influenza a virus M6PR HA membrane fusion late endosome

Role of feline ANP32 proteins in regulating polymerase activity of influenza a virus

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Journal of Integrative Agriculture 2024年第9期23卷 3145-3158页

作者： Gang Lu Feiyan Zheng Yuqing Xiao Ran Shao Jiajun Ou Xin Yin Shoujun Li Guihong Zhang College of Veterinary Medicine South China Agricultural UniversityGuangzhou 510642China State Key Laboratory for Animal Disease Control and Prevention Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150000China Maoming Branch Guangdong Laboratory for Lingnan Modern AgricultureMaoming 525000China Guangdong Laboratory for Lingnan Modern Agriculture Guangzhou 510642China

Recently,increasing natural infection cases and experimental animal challenge studies demonstrated domestic cats are susceptible to multiple subtypes influenza a virus(IAV)***,some subtype IAV strains could circulate in domestic cats after cross-species transmission and even infected humans,posing a threat to public *** factors related to viral polymerase activity could determine host range of IAV and acidic nuclear phosphoprotein 32(ANP32)is the most important one among ***,role of cat-derived ANP32 on viral polymerase activity and host range of IAV is still *** the present study,a total of 10 feline ANP32(feANP32)splice variants(including 5 feANP32A,3 feANP32B,and 2 feANP32E)were obtained from domestic cats by *** alignment results demonstrated amino acid deletions and/or insertions occurred among feANP32 variants,but all feANP32 proteins were primarily localized to cell *** replication systems for several representative IAV strains were established and the support ability of feANP32 on IAV polymerase activity was *** results indicated that most feANP32A and feANP32B splice variants were able to support all the tested IAV strains,though the support activity of a single feANP32 protein on polymerase activity varied among different IAV *** addition,the role of feANP32 in supporting H3N2 canine influenza virus was determined by investigating viral replication in ***,our study systematically investigated the support activity of feANP32 on IAV,providing a clue for further exploring the mechanism of susceptibility of cats to IAV.

关键词： ANP32 influenza a virus feline cross-species transmission

Pudilan Xiaoyan oral liquid regulates tissue inflammation and apoptosis in mice with influenza virus pneumonia

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Traditional Medicine Research 2024年第6期9卷 55-63页

作者： Shan Cao Shuang-Rong Gao Chen Ni Zi-Han Geng Ying-Li Xu Bo Pang Meng-Ping Chen Yu Zhang Shan-Shan Guo Yu-Jing Shi Li-Qi Ni Kun Wang Rong-Hua Zhao Xiao-Lan Cui Yan-Yan Bao Institute of Chinese Materia Medica China Academy of Chinese Medical SciencesBeijing 100700China Jumpcan Pharmaceutical Group Co. Ltd.Taizhou 225400China

Background:The influenza a virus is the primary cause of respiratory infections and poses a global health *** Xiaoyan oral liquid(PDL)exhibits anti-inflammatory and immunomodulatory *** is commonly employed in clinical practice to manage upper respiratory tract ***,there is still much to uncover regarding its potential therapeutic ***:Institute of cancer research mice were infected with influenza a virus via nasal *** general state of the mice,lung index,and lung index inhibition rate were used to evaluate the efficacy of ***-linked immunosorbent assay,western blotting,and immunohistochemistry were used to observe the presence of proteins and cytokines in the lung *** was evaluated using the TUNEL ***:PDL improved the mental state of influenza a virus-infected mice,reduced the lung index,and inhibited viral *** expression of interleukin-1βand tumor necrosis factor-αwere decreased,whereas the expression of interleukin-10 in the lung tissue was increased due to PDL *** addition,PDL treatment modulated Toll-like receptor 4 and MyD88 expressions in the lung *** significantly reduced apoptosis and decreased cleaved caspase-3 and PARP levels,whereas increased B-cell lymphoma-2 expression in the lung ***,the moderate-dose group of PDL exhibited a more pronounced *** findings indicate that PDL exerts a protective effect against pneumonia injury in influenza a virus-infected ***:PDL inhibited the inflammatory response and regulated apoptosis by regulating Toll-like receptor 4 and MyD88 protein expressions,thereby protecting the lung tissue from viral infection-induced lung tissue injury.

关键词： Pudilan Xiaoyan oral liquid influenza a virus immune response apoptosis Toll-like receptors

influenza a virus use of BinCARD1 to facilitate the binding of viral NP to importinα7 is counteracted by TBK1-p62 axis-mediated autophagy

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Cellular & Molecular Immunology 2022年第10期19卷 1168-1184页

作者： Xuyuan Wang Li Jiang Guangwen Wang Wenjun Shi Yuzhen Hu Bo Wang Xianying Zeng Guobin Tian Guohua Deng Jianzhong Shi Liling Liu Chengjun Li Hualan Chen College of Veterinary Medicine Gansu Agricultural UniversityLanzhou730070China Guangdong Laboratory for Lingnan Modern Agriculture Guangzhou510642China State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin150069China

As a major component of the viral ribonucleoprotein(vRNP)complex in influenza a virus(IAV),nucleoprotein(NP)interacts with isoforms of importinαfamily members,leading to the import of itself and vRNP complex into the nucleus,a process pivotal in the replication cycle of *** this study,we found that BinCARD1,an isoform of Bcl10-interacting protein with CARD(BinCARD),was leveraged by IAV for efficient viral ***1 promoted the nuclear import of the vRNP complex and newly synthesized NP and thus enhanced vRNP complex ***,we found that BinCARD1 interacted with NP to promote NP binding to importinα7,an adaptor in the host nuclear import ***,we also found that BinCARD1 promoted RIG-I-mediated innate immune signaling by mediating Lys63-linked polyubiquitination of TRAF3,and that TBK1 appeared to degrade *** showed that BinCARD1 was polyubiquitinated at residue K103 through a Lys63 linkage,which was recognized by the TBK1-p62 axis for autophagic ***,our data demonstrate that IAV leverages BinCARD1 as an important host factor that promotes viral replication,and two mechanisms in the host defense system are triggered—innate immune signaling and autophagic degradation—to mitigate the promoting effect of BinCARD1 on the life cycle of IAV.

关键词： influenza a virus BinCARD1 TRAF3 TBK1 p62

Generation and application of two monoclonal antibodies targeting conserved linear epitopes in the NP protein of influenza a virus

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Journal of Integrative Agriculture 2022年第7期21卷 2095-2105页

作者： ZHAO Yu-hui WEN Xia LI Qi-bing JIANG Li WANG Guang-wen LIANG Li-bin WANG Xiu-rong CHEN Hua-lan LI Cheng-jun State Key Laboratory of Veterinary Biotechnology Harbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069P.R.China

Monoclonal antibodies(mAbs) are widely used in virus research and disease diagnosis. The nucleoprotein(NP) of influenza a virus(IAV) plays important roles in multiple stages of the virus life cycle. Therefore, generating conserved mAbs against NP and characterizing their properties will provide useful tools for IAV research. In this study, two mAbs against the NP protein, 10 E9 and 3 F3, were generated with recombinant truncated NP proteins(NP-1 and NP-2) as immunogens. The heavy-chain subclass of both 10 E9 and 3 F3 was determined to be IgG2α, and the light-chain type was κ. Truncation and site-specific mutation analyses showed that the epitopes of mAbs 10 E9 and 3 F3 were located in the N terminal 84–89 amino acids and the C terminal 320–324 amino acids of the NP protein, respectively. We found that mAbs 10 E9 and 3 F3 reacted well with the NP protein of H1–H15 subtypes of IAV. Both 10 E9 and 3 F3 can be used in immunoprecipitation assay, and 10 E9 was also successfully applied in confocal microscopy. Furthermore, we found that the 10 E9-recognized _(84) SAGKDP_(89) epitope and 3 F3-recognized 320 ENPAH324 epitope were highly conserved in NP among all avian and human IAVs. Thus, the two mAbs we developed could be used as powerful tools in the development of diagnostic methods of IAV, and also surely promote the basic research in understanding the replication mechanisms of IAV.

关键词： influenza a virus nucleoprotein monoclonal antibody application

Discovery of the anti-influenza a virus activity of SB216763 and cyclosporine A by mining infected cells and compound cellular signatures

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Chinese Chemical Letters 2022年第5期33卷 2541-2544页

作者： Ke Tang You Wu Shubing Chen Yijing Xin Ying Guo State Key Laboratory of Bioactive Substance and Function of Natural Medicines Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China Department of Pharmacology Institute of Materia MedicaChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100050China

In this study,SB216763 and cyclosporine A were identified as anti-influenza a virus(IAV)agents by tran-scriptome signature reversion(TSR)analysis through deep mining of the cellular transcriptome of hu-man airway and lung cell lines infected with 3 strains of IAV and the chemical perturbations library.A synergistic effect of SB216763 and cyclosporine A against influenza A was disclosed by quantification of the network-based relationship,which was validated in *** with burgeoning omics approaches,transcriptome-based drug development is flourishing,which provides a novel insight into antivirals dis-covery with comprehensive cellular transcriptional information of disease and chemical perturbations in multicomponent *** strategy can be applied as a new approach in discovering multitar-get antiviral agents from approved drugs,clinical compounds,natural products or other known bioactive compounds.

关键词： Chemical perturbagen signature Host factor Transcriptome signature reversion influenza a virus SB216763 Cyclosporine A Drug combination

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 Department of Biochemistry and Molecular Biology Guangdong Medical CollegeZhanjiang 524023 China Department of Clinical Laboratory Hejin People's HospitalHejin 043300 China

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

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