42 results on '"Honglei Sun"'
Search Results
2. SRSF5-Mediated Alternative Splicing of M Gene is Essential for Influenza A Virus Replication: A Host-Directed Target Against Influenza Virus
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Qiuchen Li, Zhimin Jiang, Shuning Ren, Hui Guo, Zhimin Song, Saini Chen, Xintao Gao, Fanfeng Meng, Junda Zhu, Litao Liu, Qi Tong, Honglei Sun, Yipeng Sun, Juan Pu, Kin‐Chow Chang, and Jinhua Liu
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General Chemical Engineering ,General Engineering ,General Physics and Astronomy ,Medicine (miscellaneous) ,Virus Replication ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Mice ,Alternative Splicing ,Influenza A virus ,Influenza, Human ,Animals ,Humans ,General Materials Science ,RNA, Messenger - Abstract
Splicing of influenza A virus (IAV) RNA is an essential process in the viral life cycle that involves the co-opting of host factors. Here, it is demonstrated that induction of host serine and arginine-rich splicing factor 5 (SRSF5) by IAV facilitated viral replication by enhancing viral M mRNA splicing. Mechanistically, SRSF5 with its RRM2 domain directly bounds M mRNA at conserved sites (M mRNA position 163, 709, and 712), and interacts with U1 small nuclear ribonucleoprotein (snRNP) to promote M mRNA splicing and M2 production. Mutations introduced to the three binding sites, without changing amino acid code, significantly attenuates virus replication and pathogenesis in vivo. Likewise, SRSF5 conditional knockout in the lung protects mice against lethal IAV challenge. Furthermore, anidulafungin, an approved antifungal drug, is identified as an inhibitor of SRSF5 that effectively blocks IAV replication in vitro and in vivo. In conclusion, SRSF5 as an activator of M mRNA splicing promotes IAV replication and is a host-derived antiviral target.
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- 2022
3. Mink is a highly susceptible host species to circulating human and avian influenza viruses
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Qi Tong, Yuhai Bi, Qingzhi Liu, Xiangjun Du, Fangtao Li, Xin Zhang, Jinhua Liu, Yipeng Sun, Haoran Sun, Yuelong Shu, Yuhong Duan, Honglei Sun, Chong Li, Yongqiang Wang, Litao Liu, Juan Pu, Kin-Chow Chang, Jiyu Liu, Jianyong Du, and Jizhe Yang
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Mink (Mustelidae) ,human influenza virus ,0301 basic medicine ,Epidemiology ,animal diseases ,viruses ,030106 microbiology ,Immunology ,Reassortment ,avian influenza virus ,Biology ,Influenza A Virus, H7N9 Subtype ,medicine.disease_cause ,Microbiology ,03 medical and health sciences ,Influenza A Virus, H1N1 Subtype ,Orthomyxoviridae Infections ,Neutralization Tests ,Virology ,biology.animal ,Drug Discovery ,Pandemic ,Influenza A Virus, H9N2 Subtype ,medicine ,Animals ,Humans ,Mink ,Avian influenza virus ,Influenza A Virus, H3N2 Subtype ,Pandemic influenza ,food and beverages ,virus diseases ,General Medicine ,Influenza A virus subtype H5N1 ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,Influenza A virus ,Susceptible individual ,surveillance ,reassortment ,Parasitology ,Reassortant Viruses ,Research Article - Abstract
Pandemic influenza, typically caused by the reassortment of human and avian influenza viruses, can result in severe or fatal infections in humans. Timely identification of potential pandemic viruses must be a priority in influenza virus surveillance. However, the range of host species responsible for the generation of novel pandemic influenza viruses remains unclear. In this study, we conducted serological surveys for avian and human influenza virus infections in farmed mink and determined the susceptibility of mink to prevailing avian and human virus subtypes. The results showed that farmed mink were commonly infected with human (H3N2 and H1N1/pdm) and avian (H7N9, H5N6, and H9N2) influenza A viruses. Correlational analysis indicated that transmission of human influenza viruses occurred from humans to mink, and that feed source was a probable route of avian influenza virus transmission to farmed mink. Animal experiments showed that mink were susceptible and permissive to circulating avian and human influenza viruses, and that human influenza viruses (H3N2 and H1N1/pdm), but not avian viruses, were capable of aerosol transmission among mink. These results indicate that farmed mink could be highly permissive “mixing vessels” for the reassortment of circulating human and avian influenza viruses. Therefore, to reduce the risk of emergence of novel pandemic viruses, feeding mink with raw poultry by-products should not be permitted, and epidemiological surveillance of influenza viruses in mink farms should be urgently implemented.
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- 2021
4. p21 restricts influenza A virus by perturbing the viral polymerase complex and upregulating type I interferon signaling
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Chao Ma, Yuhan Li, Yanan Zong, Tony Velkov, Chenxi Wang, Xinyu Yang, Ming Zhang, Zhimin Jiang, Haoran Sun, Qi Tong, Honglei Sun, Juan Pu, Munir Iqbal, Jinhua Liu, Chongshan Dai, and Yipeng Sun
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A549 Cells ,Influenza A virus ,Virology ,Immunology ,Influenza, Human ,Interferon Type I ,Genetics ,Humans ,Parasitology ,Virus Replication ,Molecular Biology ,Microbiology ,Immunity, Innate - Abstract
Many cellular genes and networks induced in human lung epithelial cells infected with the influenza virus remain uncharacterized. Here, we find that p21 levels are elevated in response to influenza A virus (IAV) infection, which is independent of p53. Silencing, pharmacological inhibition or deletion of p21 promotes virus replication in vitro and in vivo, indicating that p21 is an influenza restriction factor. Mechanistically, p21 binds to the C-terminus of IAV polymerase subunit PA and competes with PB1 to limit IAV polymerase activity. Besides, p21 promotes IRF3 activation by blocking K48-linked ubiquitination degradation of HO-1 to enhance type I interferons expression. Furthermore, a synthetic p21 peptide (amino acids 36 to 43) significantly inhibits IAV replication in vitro and in vivo. Collectively, our findings reveal that p21 restricts IAV by perturbing the viral polymerase complex and activating the host innate immune response, which may aid the design of desperately needed new antiviral therapeutics.
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- 2021
5. H9N2 virus-derived M1 protein promotes H5N6 virus release in mammalian cells: Mechanism of avian influenza virus inter-species infection in humans
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Chenxi Wang, Yinghui Yu, Zhimin Jiang, Juan Pu, Xin Zhang, Fangtao Li, Jinhua Liu, Chuankuo Zhao, Haoran Sun, Jizhe Yang, Honglei Sun, Yipeng Sun, Jiyu Liu, Kin-Chow Chang, and Perez, Daniel R.
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RNA viruses ,Influenza Viruses ,Physiology ,viruses ,Cell Membranes ,M1 protein ,Pathology and Laboratory Medicine ,medicine.disease_cause ,Biochemistry ,Viral Zoonoses ,Immune Physiology ,Medicine and Health Sciences ,Influenza A Virus, H9N2 Subtype ,Biology (General) ,Virus Release ,Immune System Proteins ,biology ,virus diseases ,Precipitation Techniques ,Medical Microbiology ,Influenza A virus ,Viral Pathogens ,Viruses ,Cellular Structures and Organelles ,Pathogens ,GNB1 ,Reassortant Viruses ,Research Article ,QH301-705.5 ,Immunology ,Hemagglutinin (influenza) ,Transfection ,Research and Analysis Methods ,Microbiology ,Antibodies ,Virus ,Viral Matrix Proteins ,Virology ,Influenza, Human ,medicine ,Genetics ,Immunoprecipitation ,Animals ,Humans ,Protein Interactions ,Molecular Biology Techniques ,Microbial Pathogens ,Molecular Biology ,Viral matrix protein ,Organisms ,Biology and Life Sciences ,Membrane Proteins ,Proteins ,Cell Biology ,RC581-607 ,Viral Replication ,Influenza A virus subtype H5N1 ,Viral replication ,Influenza in Birds ,biology.protein ,Parasitology ,Immunologic diseases. Allergy ,Chickens ,Orthomyxoviruses - Abstract
H5N6 highly pathogenic avian influenza virus (HPAIV) clade 2.3.4.4 not only exhibits unprecedented intercontinental spread in poultry, but can also cause serious infection in humans, posing a public health threat. Phylogenetic analyses show that 40% (8/20) of H5N6 viruses that infected humans carried H9N2 virus-derived internal genes. However, the precise contribution of H9N2 virus-derived internal genes to H5N6 virus infection in humans is unclear. Here, we report on the functional contribution of the H9N2 virus-derived matrix protein 1 (M1) to enhanced H5N6 virus replication capacity in mammalian cells. Unlike H5N1 virus-derived M1 protein, H9N2 virus-derived M1 protein showed high binding affinity for H5N6 hemagglutinin (HA) protein and increased viral progeny particle release in different mammalian cell lines. Human host factor, G protein subunit beta 1 (GNB1), exhibited strong binding to H9N2 virus-derived M1 protein to facilitate M1 transport to budding sites at the cell membrane. GNB1 knockdown inhibited the interaction between H9N2 virus-derived M1 and HA protein, and reduced influenza virus-like particles (VLPs) release. Our findings indicate that H9N2 virus-derived M1 protein promotes avian H5N6 influenza virus release from mammalian, in particular human cells, which could be a major viral factor for H5N6 virus cross-species infection., Author summary H9N2 avian influenza viruses (AIVs), through reassortment of their internal genes with other circulating AIVs subtypes (H5N6, H7N9, H10N8 and H10N3 viruses), are known to enable the resulting novel reassortants to infect humans. However, the precise roles of H9N2 virus-derived internal genes in human infection remain unknown. Here, we found that H9N2 virus-derived (but not H5N1 virus-derived) M1 protein showed strong interaction with host GNB1 protein to enhance M1 protein transport to budding sites at the cell membrane to promote progeny virus release, hence facilitating H5N6 reassortants to infect human cells.
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- 2021
6. Canine Influenza Virus A(H3N2) Clade with Antigenic Variation, China, 2016–2017
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Guoxia Bing, Zhaofei Xia, Liwei Zhou, Shikai Song, Junyi Hu, Juan Pu, Mingyang Wang, Honglei Sun, Haoran Sun, Mingyue Chen, Qian Wang, Yipeng Sun, Yanli Lyu, and Jinhua Liu
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dogs ,Epidemiology ,viruses ,Canine influenza ,lcsh:Medicine ,serology ,Canine Influenza Virus A(H3N2) Clade with Antigenic Variation, China, 2016–2017 ,adaptation ,medicine.disease_cause ,Serology ,0302 clinical medicine ,Zoonoses ,canine influenza ,Influenza A virus ,Dog Diseases ,030212 general & internal medicine ,Clade ,Phylogeny ,Zoonotic Infection ,mammalian adaptations ,Dispatch ,virus diseases ,H3N2 ,Antigenic Variation ,Infectious Diseases ,antigenicity ,surveillance ,influenza ,Microbiology (medical) ,China ,030231 tropical medicine ,Biology ,Virus ,lcsh:Infectious and parasitic diseases ,respiratory infections ,03 medical and health sciences ,Orthomyxoviridae Infections ,Influenza, Human ,evolution ,medicine ,Antigenic variation ,Animals ,Humans ,influenza A virus ,lcsh:RC109-216 ,Influenza A Virus, H3N2 Subtype ,lcsh:R ,Virology ,PB2 - Abstract
During 2012–2017, we collected throat swabs from dogs in China to characterize canine influenza virus (CIV) A(H3N2) isolates. A new antigenically and genetically distinct CIV H3N2 clade possessing mutations associated with mammalian adaptation emerged in 2016 and replaced previously circulating strains. This clade probably poses a risk for zoonotic infection.
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- 2019
7. Swine MicroRNAs ssc-miR-221-3p and ssc-miR-222 Restrict the Cross-Species Infection of Avian Influenza Virus
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Hsiang-Yu Yuan, Junda Zhu, Zhimin Jiang, Yipeng Sun, Honglei Sun, Tong Wang, Juan Pu, Jingwei Song, Weihua Gao, Haoran Sun, Jinhua Liu, and Chenxi Wang
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Swine ,Immunology ,Biology ,Virus Replication ,medicine.disease_cause ,Microbiology ,Virus ,Birds ,Cell membrane ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Virology ,Macrophages, Alveolar ,microRNA ,medicine ,Influenza A virus ,Animals ,Humans ,030304 developmental biology ,Homeodomain Proteins ,0303 health sciences ,Host (biology) ,Gene Expression Profiling ,virus diseases ,Virus-Cell Interactions ,Up-Regulation ,MicroRNAs ,HEK293 Cells ,medicine.anatomical_structure ,Apoptosis ,Influenza in Birds ,030220 oncology & carcinogenesis ,Insect Science ,Host-Pathogen Interactions ,Alveolar macrophage - Abstract
Avian influenza virus (AIV) can cross species barriers to infect humans and other mammals. However, these species-cross transmissions are most often dead-end infections due to host restriction. Current research about host restriction focuses mainly on the barriers of cell membrane, nuclear envelope, and host proteins; whether microRNAs (miRNAs) play a role in host restriction is largely unknown. In this study, we used porcine alveolar macrophage (PAM) cells as a model to elucidate the role of miRNAs in host range restriction. During AIV infection, 40 dysregulation expressed miRNAs were selected in PAM cells. Among them, two Sus scrofa (ssc; swine) miRNAs, ssc-miR-221-3p and ssc-miR-222, could inhibit the infection and replication of AIV in PAM cells by directly targeting viral genome and inducing cell apoptosis via inhibiting the expression of anti-apoptotic protein HMBOX1. Avian but not swine influenza virus caused upregulated expressions of ssc-miR-221-3p and ssc-miR-222 in PAM cells. We further found that NF-κB P65 was more effectively phosphorylated upon AIV infection and that P65 functioned as a transcription activator to regulate the AIV-induced expression of miR-221-3p/222. Importantly, we found that ssc-miR-221-3p and ssc-miR-222 could also be specifically upregulated upon AIV infection in newborn pig tracheal epithelial (NPTr) cells and also exerted anti-AIV function. In summary, our study indicated that miRNAs act as a host barrier during cross-species infection of influenza A virus. IMPORTANCE The host range of an influenza A virus is determined by species-specific interactions between virus and host cell factors. Host miRNAs can regulate influenza A virus replication; however, the role of miRNAs in host species specificity is unclear. Here, we show that the induced expression of ssc-miR-221-3p and ssc-miR-222 in swine cells is modulated by NF-κB P65 phosphorylation in response to AIV infection but not swine influenza virus infection. ssc-miR-221-3p and ssc-miR-222 exerted antiviral function via targeting viral RNAs and causing apoptosis by inhibiting the expression of HMBOX1 in host cells. These findings uncover miRNAs as a host range restriction factor that limits cross-species infection of influenza A virus.
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- 2020
8. Truncation of PA-X Contributes to Virulence and Transmission of H3N8 and H3N2 Canine Influenza Viruses in Dogs
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Yipeng Sun, Litao Liu, Chao Ma, Honglei Sun, Jinhua Liu, Qi Tong, Ye Shen, Juan Pu, Tong Wang, Shikai Song, and Munir Iqbal
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Canine influenza ,viruses ,Immunology ,Virulence ,Biology ,Viral Nonstructural Proteins ,medicine.disease_cause ,Virus Replication ,Microbiology ,Virus ,Madin Darby Canine Kidney Cells ,Influenza A Virus, H3N8 Subtype ,Plasmid ,Dogs ,Orthomyxoviridae Infections ,Virology ,Influenza A virus ,medicine ,Animals ,Humans ,Viral shedding ,Gene ,Influenza A Virus, H3N2 Subtype ,Virus Shedding ,Repressor Proteins ,HEK293 Cells ,Viral replication ,Insect Science ,Pathogenesis and Immunity - Abstract
Equine-origin H3N8 and avian-origin H3N2 canine influenza viruses (CIVs) prevalent in dogs are thought to pose a public health threat arising from intimate contact between dogs and humans. However, our understanding of CIV virulence is still limited. Influenza A virus PA-X is a fusion protein encoded in part by a +1 frameshifted open reading frame (X-ORF) in segment 3. The X-ORF can be translated in full-length (61-amino-acid) or truncated (41-amino-acid) form. Genetic analysis indicated that the X-ORFs of equine H3N8 and avian H3N2 influenza viruses encoded 61 amino acids but were truncated after introduction into dogs. To determine the effect of PA-X truncation on the biological characteristics of CIVs, we constructed four recombinant viruses on H3N8 and H3N2 CIV backgrounds bearing truncated or full-length PA-Xs. We observed that truncation of PA-X increased growth of both H3N8 and H3N2 CIVs in MDCK cells and suppressed expression from cotransfected plasmids in MDCK cells. Furthermore, truncation of PA-X enhanced viral pathogenicity in dogs, as shown by aggravated clinical symptoms and histopathological changes, increased viral replication in the respiratory system, and prolonged virus shedding. Additionally, CIVs with truncated PA-Xs were transmitted more efficiently in dogs. Global gene expression profiling of the lungs of infected dogs revealed that differentially expressed genes were mainly associated with inflammatory responses, which might contribute to the pathogenicity of PA-X-truncated CIVs. Our findings revealed that truncation of PA-X might be important for the adaptation of influenza viruses to dogs. IMPORTANCE Epidemics of equine-origin H3N8 and avian-origin H3N2 influenza viruses in canine populations are examples of successful cross-species transmission of influenza A viruses. Genetic analysis showed that the PA-X genes of equine H3N8 or avian H3N2 influenza viruses were full-length, with X-ORFs encoding 61 amino acids; however, those of equine-origin H3N8 or avian-origin H3N2 CIVs were truncated, suggesting that PA-X truncation occurred after transmission to dogs. In this study, we extended the PA-X genes of H3N8 and H3N2 CIVs and compared the biological characteristics of CIVs bearing different lengths of PA-X. We demonstrated that for both H3N8 and H3N2 viruses, truncation of PA-X increased virus yields in MDCK cells and enhanced viral replication, pathogenicity, and transmission in dogs. These results might reflect enhanced suppression of host gene expression and upregulation of genes related to inflammatory responses. Collectively, our data partially explain the conservation of truncated PA-X in CIVs.
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- 2020
9. IFI16 directly senses viral RNA and enhances RIG-I transcription and activation to restrict influenza virus infection
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Weihua Gao, Kin-Chow Chang, Chengjiang Gao, Shufang Yu, Honglei Sun, Juan Pu, Jinhua Liu, Zhimin Jiang, Yuying Zhang, Qi Tong, Fanhua Wei, Yipeng Sun, Mingyang Wang, and Tong Wang
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Microbiology (medical) ,viruses ,Immunology ,RNA polymerase II ,chemical and pharmacologic phenomena ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Microbiology ,Virus ,03 medical and health sciences ,Mice ,Orthomyxoviridae Infections ,Transcription (biology) ,Influenza A virus ,medicine ,Genetics ,Animals ,Humans ,Receptors, Immunologic ,Promoter Regions, Genetic ,Gene ,030304 developmental biology ,0303 health sciences ,Innate immune system ,biology ,030306 microbiology ,RIG-I ,RNA ,virus diseases ,Nuclear Proteins ,RNA-Binding Proteins ,Cell Biology ,biochemical phenomena, metabolism, and nutrition ,Phosphoproteins ,Immunity, Innate ,Cell biology ,Up-Regulation ,Host-Pathogen Interactions ,Interferon Type I ,biology.protein ,DEAD Box Protein 58 ,RNA, Viral ,biological phenomena, cell phenomena, and immunity ,Protein Binding ,Signal Transduction - Abstract
The retinoic acid-inducible gene I (RIG-I) receptor senses cytoplasmic viral RNA and activates type I interferons (IFN-I) and downstream antiviral immune responses. How RIG-I binds to viral RNA and how its activation is regulated remains unclear. Here, using IFI16 knockout cells and p204-deficient mice, we demonstrate that the DNA sensor IFI16 enhances IFN-I production to inhibit influenza A virus (IAV) replication. IFI16 positively upregulates RIG-I transcription through direct binding to and recruitment of RNA polymerase II to the RIG-I promoter. IFI16 also binds to influenza viral RNA via its HINa domain and to RIG-I protein with its PYRIN domain, thus promoting IAV-induced K63-linked polyubiquitination and RIG-I activation. Our work demonstrates that IFI16 is a positive regulator of RIG-I signalling during influenza virus infection, highlighting its role in the RIG-I-like-receptor-mediated innate immune response to IAV and other RNA viruses, and suggesting its possible exploitation to modulate the antiviral response.
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- 2020
10. An R195K Mutation in the PA-X Protein Increases the Virulence and Transmission of Influenza A Virus in Mammalian Hosts
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Yipeng Sun, Mingyue Chen, Yuhai Bi, Zhen Wang, Juan Pu, Xuxiao Zhang, Jinhua Liu, Zhe Hu, Guanlong Xu, Mingyang Wang, Honglei Sun, Munir Iqbal, and Qi Tong
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Virulence Factors ,viruses ,Immunology ,Reassortment ,Mutation, Missense ,Virulence ,Cross-species transmission ,Biology ,Viral Nonstructural Proteins ,medicine.disease_cause ,Microbiology ,Virus ,Madin Darby Canine Kidney Cells ,Dogs ,Virology ,Pandemic ,Influenza, Human ,Influenza A virus ,medicine ,Animals ,Humans ,Transmission (medicine) ,virus diseases ,Influenza A virus subtype H5N1 ,Repressor Proteins ,HEK293 Cells ,Amino Acid Substitution ,A549 Cells ,Insect Science ,Pathogenesis and Immunity - Abstract
In the 21st century, the emergence of H7N9 and H1N1/2009 influenza viruses, originating from animals and causing severe human infections, has prompted investigations into the genetic alterations required for cross-species transmission. We previously found that replacement of the human-origin PA gene segment in avian influenza virus (AIV) could overcome barriers to cross-species transmission. Recently, it was reported that the PA gene segment encodes both the PA protein and a second protein, PA-X. Here, we investigated the role of PA-X. We found that an H9N2 avian influenza reassortant virus bearing a human-origin H1N1/2009 PA gene was attenuated in mice after the loss of PA-X. Reverse genetics analyses of PA-X substitutions conserved in human influenza viruses indicated that R195K, K206R, and P210L substitutions conferred significantly increased replication and pathogenicity on H9N2 virus in mice and ferrets. PA-X R195K was present in all human H7N9 and H1N1/2009 viruses and predominated in human H5N6 viruses. Compared with PA-X 195R, H7N9 influenza viruses bearing PA-X 195K showed increased replication and transmission in ferrets. We further showed that PA-X 195K enhanced lung inflammatory responses, potentially due to decreased host shutoff function. A competitive transmission study in ferrets indicated that 195K provides a replicative advantage over 195R in H1N1/2009 viruses. In contrast, PA-X 195K did not influence the virulence of H9N2 AIV in chickens, suggesting that the effects of the substitution were mammal specific. Therefore, future surveillance efforts should scrutinize this region of PA-X because of its potential impact on cross-species transmission of influenza viruses. IMPORTANCE Four influenza pandemics in humans (the Spanish flu of 1918 [H1N1], the Asian flu of 1957 [H2N2], the Hong Kong flu of 1968 [H3N2], and the swine origin flu of 2009 [H1N1]) are all proposed to have been caused by avian or swine influenza viruses that acquired virulence factors through adaptive mutation or reassortment with circulating human viruses. Currently, influenza viruses circulating in animals are repeatedly transmitted to humans, posing a significant threat to public health. However, the molecular properties accounting for interspecies transmission of influenza viruses remain unclear. In the present study, we demonstrated that PA-X plays an important role in cross-species transmission of influenza viruses. At least three human-specific amino acid substitutions in PA-X dramatically enhanced the adaptation of animal influenza viruses in mammals. In particular, PA-X 195K might have contributed to cross-species transmission of H7N9, H5N6, and H1N1/2009 viruses from animal reservoirs to humans.
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- 2019
11. Genetic evolution of influenza H9N2 viruses isolated from various hosts in China from 1994 to 2013
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Zejiang Wang, Gang Wu, Chenxi Wang, Guoxia Bing, Robert A. Carter, Jinliang Wang, Juan Pu, Shuoguo Wang, Jinhua Liu, Yipeng Sun, Honglei Sun, Chong Li, Yongqiang Wang, Lan Wang, and Robert G. Webster
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0301 basic medicine ,China ,Genotype ,genetic evolution ,Epidemiology ,viruses ,Immunology ,host range ,Zoology ,Genome, Viral ,Biology ,medicine.disease_cause ,Microbiology ,H5N1 genetic structure ,Host Specificity ,H9N2 influenza virus ,Virus ,Birds ,Evolution, Molecular ,03 medical and health sciences ,Orthomyxoviridae Infections ,Virology ,geographic distribution ,Drug Discovery ,Evolution of influenza ,Influenza A Virus, H9N2 Subtype ,Influenza A virus ,medicine ,Animals ,Humans ,Clade ,Mammals ,Molecular Epidemiology ,Molecular epidemiology ,Host (biology) ,Genetic Variation ,Sequence Analysis, DNA ,General Medicine ,Phylogeography ,030104 developmental biology ,Infectious Diseases ,Viral evolution ,Original Article ,Parasitology - Abstract
Influenza H9N2 subtype viruses and their reassortants (such as H7N9) are posing increasing threats to birds and humans in China. During 2009–2013, multiple novel subtype viruses with H9N2 original genes emerged in China. Yet, the genetic evolution of H9N2 viruses in various host organisms in China has not been systematically investigated since 2009. In the present study, we performed large-scale sequence analysis of H9N2 viral genomes from public databases, representing the spectrum of viruses isolated from birds, mammals and humans in China from 1994 to 2013, and updated the clade classification for each segment. We identified 117 distinct genotypes in 730 H9N2 viruses. We analyzed the sequences of all eight segments in each virus and found three important time points: the years 2000, 2006 and 2010. In the periods divided by these years, genotypic diversity, geographic distribution and host range changed considerably. Genotypic diversity fluctuated greatly in 2000 and 2006. Since 2010, a single genotype became predominant in poultry throughout China, and the eastern coastal region became the newly identified epidemic center. Throughout their 20-year prevalence in China, H9N2 influenza viruses have emerged and adapted from aquatic birds to chickens. The minor avian species and wild birds exacerbated H9N2 genotypes by providing diversified genes, and chickens were the most prevalent vector in which the viruses evolved and expanded their prevalence. It is the necessity for surveillance and disease control on live-bird markets, poultry farms and wild-bird habitats in China.
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- 2017
12. Nested RT-PCR method for the detection of European avian-like H1 swine influenza A virus
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Jinhua Liu, Yuan Zhang, Honglei Sun, Wei Yandi, Juan Pu, Chen-fang Yu, and Pei Xingyao
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0301 basic medicine ,European avian-like H1 ,Nested rt pcr ,HA gene ,Agriculture (General) ,viruses ,Plant Science ,Biology ,medicine.disease_cause ,Biochemistry ,S1-972 ,nested RT-PCR ,03 medical and health sciences ,swine influenza A virus ,Food Animals ,Pandemic ,molecular diagnosis ,Influenza A virus ,medicine ,Multiplex ,Gene ,Plaque-forming unit ,Ecology ,Virology ,Reverse transcriptase ,030104 developmental biology ,Animal Science and Zoology ,Primer (molecular biology) ,Agronomy and Crop Science ,Food Science - Abstract
Swine influenza A virus (swine IAV) circulates worldwide in pigs and poses a serious public health threat, as evidenced by the 2009 H1N1 influenza pandemic. Among multiple subtypes/lineages of swine influenza A viruses, European avian-like (EA) H1N1 swine IAV has been dominant since 2005 in China and caused infections in humans in 2010. Highly sensitive and specific methods of detection are required to differentiate EA H1N1 swine IAVs from viruses belonging to other lineages and subtypes. In this study, a nested reverse transcription (RT)-PCR assay was developed to detect EA H1 swine IAVs. Two primer sets (outer and inner) were designed specifically to target the viral hemagglutinin genes. Specific PCR products were obtained from all tested EA H1N1 swine IAV isolates, but not from other lineages of H1 swine IAVs, other subtypes of swine IAVs, or other infectious swine viruses. The sensitivity of the nested RT-PCR was improved to 1 plaque forming unit (PFU) mL −1 which was over 10 4 PFU mL −1 for a previously established multiplex RT-PCR method. The nested RT-PCR results obtained from screening 365 clinical samples were consistent with those obtained using conventional virus isolation methods combined with sequencing. Thus, the nested RT-PCR assay reported herein is more sensitive and suitable for the diagnosis of clinical infections and surveillance of EA H1 swine IAVs in pigs and humans.
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- 2016
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13. Induction of PGRN by influenza virus inhibits the antiviral immune responses through downregulation of type I interferons signaling
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Fanhua Wei, Zhimin Jiang, Honglei Sun, Juan Pu, Yipeng Sun, Mingyang Wang, Qi Tong, Yuhai Bi, Xiaojing Ma, George Fu Gao, and Jinhua Liu
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Male ,RNA viruses ,Influenza Viruses ,Viral Diseases ,Physiology ,Pathology and Laboratory Medicine ,Biochemistry ,Mice ,White Blood Cells ,Progranulins ,Animal Cells ,Immune Physiology ,Zoonoses ,Medicine and Health Sciences ,Small interfering RNAs ,Post-Translational Modification ,Biology (General) ,Cells, Cultured ,Mice, Knockout ,0303 health sciences ,Immune System Proteins ,030302 biochemistry & molecular biology ,NF-kappa B ,H5N1 ,Medical microbiology ,Orthomyxoviridae ,Nucleic acids ,Infectious Diseases ,Influenza A virus ,Viral Pathogens ,Host-Pathogen Interactions ,Interferon Type I ,Viruses ,Pathogens ,Cellular Types ,Signal Transduction ,Research Article ,QH301-705.5 ,Immune Cells ,Immunology ,Down-Regulation ,Transfection ,Research and Analysis Methods ,Antiviral Agents ,Microbiology ,Antibodies ,03 medical and health sciences ,Orthomyxoviridae Infections ,Virology ,Genetics ,Animals ,Molecular Biology Techniques ,Non-coding RNA ,Microbial Pathogens ,Molecular Biology ,Immune Evasion ,030304 developmental biology ,Blood Cells ,Biology and life sciences ,Macrophages ,Organisms ,Ubiquitination ,Correction ,Proteins ,Cell Biology ,RC581-607 ,Immunity, Innate ,Influenza ,Gene regulation ,RNA ,Parasitology ,Gene expression ,Immunologic diseases. Allergy ,Orthomyxoviruses - Abstract
Type I interferons (IFNs) play a critical role in host defense against influenza virus infection, and the mechanism of influenza virus to evade type I IFNs responses remains to be fully understood. Here, we found that progranulin (PGRN) was significantly increased both in vitro and in vivo during influenza virus infection. Using a PGRN knockdown assay and PGRN-deficient mice model, we demonstrated that influenza virus-inducing PGRN negatively regulated type I IFNs production by inhibiting the activation of NF-κB and IRF3 signaling. Furthermore, we showed that PGRN directly interacted with NF-κB essential modulator (NEMO) via its Grn CDE domains. We also verified that PGRN recruited A20 to deubiquitinate K63-linked polyubiquitin chains on NEMO at K264. In addition, we found that macrophage played a major source of PGRN during influenza virus infection, and PGRN neutralizing antibodies could protect against influenza virus-induced lethality in mice. Our data identify a PGRN-mediated IFN evasion pathway exploited by influenza virus with implication in antiviral applications. These findings also provide insights into the functions and crosstalk of PGRN in innate immunity., Author summary The innate immune system is the first line of host defense against microbial infection, while viruses develop several strategies to evade the host defense. It is of great significance to explore the mechanism by which viruses to evade the antiviral host defense. Previous studies have found that progranulin (PGRN) plays an important role in a variety of physiologic and disease processes. Here, we demonstrated that PGRN induced by influenza virus negatively regulated type I IFN production by inhibiting the activation of NF-κB and IRF3 signaling. We further showed that PGRN directly interacted with NEMO via its Grn CDE domains and recruited A20 to deubiquitinate K63-linked polyubiquitin chains on NEMO. Macrophage played a major source of PGRN during influenza virus infection, and PGRN neutralizing antibodies could protect against influenza virus-induced lethality in mice. Our findings highlight a new strategy whereby influenza virus to evade type I IFN-mediated antiviral immune response and also provide insights into the functions and crosstalk of PGRN in innate immunity.
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- 2019
14. The use of pyrosequencing for detection of hemagglutinin mutations associated with increased pathogenicity of H5N1 avian influenza viruses in mammals
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Mingyang Wang, Juan Pu, Honglei Sun, Chenxi Wang, Wu Shaoqiang, Yipeng Sun, Xuxiao Zhang, Jinhua Liu, Caixia Wang, Guoxia Bing, Yongning Zhang, and Xiangmei Lin
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0301 basic medicine ,animal diseases ,Virulence ,Biology ,medicine.disease_cause ,Cleavage (embryo) ,Rapid detection ,Serine ,03 medical and health sciences ,Orthomyxoviridae Infections ,medicine ,Animals ,Self-Sustained Sequence Replication ,chemistry.chemical_classification ,Mammals ,General Veterinary ,Influenza A Virus, H5N1 Subtype ,virus diseases ,Pathogenicity ,Virology ,Influenza A virus subtype H5N1 ,Amino acid ,030104 developmental biology ,Hemagglutinins ,chemistry ,Influenza A virus ,Mutation ,Pyrosequencing ,Brief Communications - Abstract
Hemagglutinin (HA) cleavage is critical for virulence of influenza viruses. The amino acid residue at the P6 position of the HA cleavage site (HACS) has been shown to be most variable and to have a direct correlation with the cleavage efficiency and pathogenicity of H5N1 avian influenza viruses (AIVs) in mammals. Among these amino acid variants, serine has been associated with the highest virulence in mammals, and its detection may serve as an indicator for H5N1 AIVs with high pathogenicity and potential public risk. We developed a rapid detection method based on reverse-transcription (RT)-PCR and pyrosequencing to detect a mutation at the HACS that is associated with increased pathogenicity of H5N1 AIVs in mammals. Herein, we provide a specific, sensitive, and reliable method for rapid detection of one of the virulence determinants associated with increased pathogenicity of H5N1 AIVs in mammals.
- Published
- 2018
15. PA-X is a virulence factor in avian H9N2 influenza virus
- Author
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Jinliang Wang, Lu Qi, Jinhua Liu, Kin-Chow Chang, Guanlong Xu, Huijie Gao, Juan Pu, Honglei Sun, Yipeng Sun, and Weili Kong
- Subjects
Virulence Factors ,animal diseases ,viruses ,Virulence ,Viral Nonstructural Proteins ,Biology ,medicine.disease_cause ,H5N1 genetic structure ,Virulence factor ,Virus ,Antigenic drift ,Cell Line ,Microbiology ,Mice ,Virology ,Influenza, Human ,Influenza A Virus, H9N2 Subtype ,medicine ,Influenza A virus ,Animals ,Humans ,Lung ,Mice, Inbred BALB C ,Viral culture ,virus diseases ,Influenza A virus subtype H5N1 ,Repressor Proteins ,Influenza in Birds ,Cytokines ,Female ,Chickens - Abstract
H9N2 influenza viruses have been circulating worldwide in multiple avian species, and regularly infect pigs and humans. Recently, a novel protein, PA-X, produced from the PA gene by ribosomal frameshifting, was demonstrated to be an antivirulence factor in pandemic 2009 H1N1, highly pathogenic avian H5N1 and 1918 H1N1 viruses. However, a similar role of PA-X in the prevalent H9N2 avian influenza viruses has not been established. In this study, we compared the virulence and cytopathogenicity of H9N2 WT virus and H9N2 PA-X-deficient virus. Loss of PA-X in H9N2 virus reduced apoptosis and had a marginal effect on progeny virus output in human pulmonary adenocarcinoma (A549) cells. Without PA-X, PA was less able to suppress co-expressed GFP in human embryonic kidney 293T cells. Furthermore, absence of PA-X in H9N2 virus attenuated viral pathogenicity in mice, which showed no mortality, reduced progeny virus production, mild-to-normal lung histopathology, and dampened proinflammatory cytokine and chemokine response. Therefore, unlike previously reported H1N1 and H5N1 viruses, we show that PA-X protein in H9N2 virus is a pro-virulence factor in facilitating viral pathogenicity and that the pro- or antivirulence role of PA-X in influenza viruses is virus strain-dependent.
- Published
- 2015
16. Enhanced pathogenicity and neurotropism of mouse-adapted H10N7 influenza virus are mediated by novel PB2 and NA mutations
- Author
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Chenxi Wang, Jinhua Liu, Yipeng Sun, Guanlong Xu, Juan Pu, Honglei Sun, Ming Jiang, Weihua Gao, Mingyang Wang, Xuxiao Zhang, and Kin-Chow Chang
- Subjects
0301 basic medicine ,Central Nervous System ,Virulence Factors ,viruses ,Adaptation, Biological ,Mutation, Missense ,Virulence ,Neuraminidase ,medicine.disease_cause ,Virus ,03 medical and health sciences ,Viral Proteins ,Orthomyxoviridae Infections ,Serial passage ,Virology ,medicine ,Animals ,Serial Passage ,Mutation ,Mice, Inbred BALB C ,biology ,virus diseases ,H10N7 influenza virus, mouse-adaptation, neurovirulence, pathogenicity ,RNA-Dependent RNA Polymerase ,Influenza A virus subtype H5N1 ,Viral Tropism ,030104 developmental biology ,Viral replication ,Influenza A virus ,biology.protein ,Tissue tropism ,Female - Abstract
The H10 subtype of avian influenza viruses (AIVs) circulates globally in wild birds and poultry, and this subtype has been shown to be increasingly prevalent in China. Among the various H10 viruses, H10N7 AIVs have caused repeated mammal and human infections. To investigate their genetic adaptation in mammals, we generated a mouse-adapted avian H10N7 variant (A/mallard/Beijing/27/2011-MA; BJ27-MA) which exhibited increased virulence in mice compared to wild-type virus and acquired neurotropism. Sequencing showed the absence of the widely recognized mammalian adaptation markers of E627K and D701N in PB2 in the mouse-adapted strain; instead, five amino acid mutations were identified: E158G and M631L in PB2; G218E in haemagglutinin (H3 numbering); and K110E and S453I in neuraminidase (NA). The neurovirulence of the BJ27-MA virus necessitated the combined presence of the PB2 and NA mutations. Mutations M631L and E158G of PB2 and K110E of NA were required to mediate increased virus replication and severity of infection in mice and mammalian cells. PB2-M631L was functionally the most dominant mutation in that it strongly upregulated viral polymerase activity and played a critical role in the enhancement of virus replication and disease severity in mice. K110E mutation in NA, on the other hand, significantly promoted NA enzymatic activity. These results indicate that the novel mutations in PB2 and NA genes are critical for the adaptation of H10N7 AIV in mice, and they could serve as molecular signatures of virus transmission to mammalian hosts, including humans.
- Published
- 2017
17. A Multiplex RT-PCR Assay for Detection and Differentiation of Avian-Origin Canine H3N2, Equine-Origin H3N8, Human-Origin H3N2, and H1N1/2009 Canine Influenza Viruses
- Author
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Yipeng Sun, Qian Wang, Honglei Sun, Chenxi Wang, Jinhua Liu, Juan Pu, and Junyi Hu
- Subjects
0301 basic medicine ,RNA viruses ,Influenza Viruses ,Canine influenza ,viruses ,lcsh:Medicine ,Artificial Gene Amplification and Extension ,medicine.disease_cause ,Pathology and Laboratory Medicine ,Polymerase Chain Reaction ,law.invention ,Geographical Locations ,law ,Medicine and Health Sciences ,Multiplex ,lcsh:Science ,Polymerase chain reaction ,Mammals ,Multidisciplinary ,biology ,H1N1 ,virus diseases ,Orthomyxoviridae ,Real-time polymerase chain reaction ,Medical Microbiology ,Influenza A virus ,Viral Pathogens ,Viruses ,Vertebrates ,Pathogens ,Research Article ,China ,Asia ,Hemagglutinin (influenza) ,Microbiology ,03 medical and health sciences ,Dogs ,Extraction techniques ,Canine Distemper Virus ,Multiplex polymerase chain reaction ,medicine ,Animals ,Humans ,Molecular Biology Techniques ,Microbial Pathogens ,Molecular Biology ,Biology and life sciences ,lcsh:R ,Organisms ,Reverse Transcriptase-Polymerase Chain Reaction ,biology.organism_classification ,Virology ,Influenza A virus subtype H5N1 ,RNA extraction ,Research and analysis methods ,030104 developmental biology ,Amniotes ,People and Places ,Paramyxoviruses ,biology.protein ,lcsh:Q ,Multiplex Polymerase Chain Reaction ,Orthomyxoviruses - Abstract
Virological and serological surveys have documented that H1N1/2009, avian-origin canine H3N2 (cH3N2), seasonal human-origin H3N2 (hH3N2), and equine-origin H3N8 influenza viruses are consistently circulating in dogs. In the present study, a multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was developed for simultaneous detection and differentiation of these influenza viruses. Four primer sets were designed to target the hemagglutinin genes of H1N1/2009, cH3N2, hH3N2, and H3N8 canine influenza viruses (CIVs). This mRT-PCR assay demonstrated high specificity and sensitivity for the four CIV subtypes. Additionally, mRT-PCR results obtained from 420 clinical samples were consistent with those obtained by the conventional virus isolation method. Our mRT-PCR assay is reliable for clinical diagnosis and rapid identification of CIVs.
- Published
- 2017
18. Isolation and characterization of H4N6 avian influenza viruses from mallard ducks in Beijing, China
- Author
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Junyi Hu, Yipeng Sun, Honglei Sun, Xinyi Xu, Jinhua Liu, Guoxia Bing, Chenxi Wang, and Juan Pu
- Subjects
RNA viruses ,0301 basic medicine ,Influenza Viruses ,viruses ,Waterfowl ,lcsh:Medicine ,Pathology and Laboratory Medicine ,medicine.disease_cause ,Poultry ,Animal Diseases ,Gene flow ,Mice ,Zoonoses ,Genotype ,Medicine and Health Sciences ,Influenza A virus ,lcsh:Science ,Phylogeny ,Data Management ,Mammals ,Multidisciplinary ,Bird Genetics ,Transmission (medicine) ,virus diseases ,Phylogenetic Analysis ,Animal Models ,Phylogenetics ,Ducks ,Infectious Diseases ,Experimental Organism Systems ,Medical Microbiology ,Viral Pathogens ,Beijing ,Vertebrates ,Viruses ,Female ,Pathogens ,Research Article ,Avian Influenza ,Computer and Information Sciences ,Guinea Pigs ,030106 microbiology ,Animals, Wild ,Genome, Viral ,Biology ,Research and Analysis Methods ,Rodents ,Microbiology ,H5N1 genetic structure ,Virus ,Birds ,Animal Influenza ,03 medical and health sciences ,Virology ,Genetics ,medicine ,Animals ,Evolutionary Systematics ,Microbial Pathogens ,Taxonomy ,Evolutionary Biology ,Body Weight ,lcsh:R ,Organisms ,Biology and Life Sciences ,Viral Replication ,Influenza A virus subtype H5N1 ,030104 developmental biology ,Viral replication ,Fowl ,Influenza in Birds ,Amniotes ,lcsh:Q ,Animal Genetics ,Zoology ,Viral Transmission and Infection ,Orthomyxoviruses - Abstract
The novel H7N9 influenza virus, which has caused severe disease in humans in China, is a reassortant with surface genes derived from influenza viruses in wild birds. This highlights the importance of monitoring influenza viruses in these hosts. However, surveillance of influenza virus in wild birds remains very limited in China. In this study, we isolated four H4N6 avian influenza viruses (AIVs) from mallard ducks in Beijing Wetland Park, which is located on the East Asia-Australasia migratory flyway. The gene segments of these Chinese H4N6 viruses were closest to AIVs in wild birds from Mongolia or the Republic of Georgia, indicating the interregional AIV gene flow among these countries. All of our isolates belonged to a novel genotype that was different from other H4N6 viruses isolated in China. We further evaluated the virulence and transmission of two representative H4N6 strains in mammalian models. We found that both of these H4N6 viruses replicated efficiently in mice without adaptation. Additionally, these two strains had a 100% transmission rate in guinea pigs via direct contact, but they had not acquired respiratory droplet transmissibility. These results reveal the potential threat to human health of H4N6 viruses in migratory birds and the need for enhanced surveillance of AIVs in wild birds.
- Published
- 2017
19. Structures and Receptor Binding of Hemagglutinins from Human-Infecting H7N9 Influenza Viruses
- Author
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Weimin Gong, Ying Wu, Hao Song, Jinghua Yan, Yanfang Zhang, Dayan Wang, Yu Wang, Feng Gao, Yuelong Shu, Joel Haywood, Fei Wang, Zheng Fan, Wenjun Liu, Wei Zhang, Honglei Sun, Jinhua Liu, Jianxun Qi, Yi Shi, Yuhai Bi, George F. Gao, and Cheng-Feng Qin
- Subjects
Protein Conformation ,Viral protein ,Mutant ,Glycine ,Hemagglutinin Glycoproteins, Influenza Virus ,Receptors, Cell Surface ,Biology ,Crystallography, X-Ray ,medicine.disease_cause ,Virus ,Birds ,Protein structure ,Influenza, Human ,medicine ,Influenza A virus ,Animals ,Humans ,Receptor ,chemistry.chemical_classification ,Multidisciplinary ,Virology ,Amino acid ,chemistry ,Influenza in Birds - Abstract
Two Viruses to Bind Structural studies of two different H7N9 influenza viruses isolated from humans—A/Shanghai/1/2013 and A/Anhui/1/2013—which have different amino acid sequences in the receptor binding site, provide data indicating that the virus is in transition with respect to host adaptation. The Shanghai virus was one of the first isolated in humans that binds avian receptor glycans with high affinity, but binds poorly to human receptors. However, the later Anhui isolates can bind both avian and human receptors at high affinity. Shi et al. (p. 243 , published online 5 September) show that four hydrophobic mutations contribute to acquisition of affinity for the human receptor by the virus hemagglutinin (HA) and confirm this effect in binding studies with virus particles. Further comparison of a mutant H7N9 A/Anhui/1/2013 HA with the bird flu H5N1 virus revealed the significance of some of the naturally occurring changes observed in circulating H7N9 viruses, which helps to explain how these viruses have been able to cause many severe human infections in a short time.
- Published
- 2013
20. Generation and protective efficacy of a cold-adapted attenuated avian H9N2 influenza vaccine
- Author
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Wei Yandi, Jinhua Liu, Lu Qi, Honglei Sun, Juan Pu, Yipeng Sun, and Huijie Gao
- Subjects
CD4-Positive T-Lymphocytes ,0301 basic medicine ,Influenza vaccine ,animal diseases ,viruses ,030106 microbiology ,CD8-Positive T-Lymphocytes ,Biology ,Vaccines, Attenuated ,medicine.disease_cause ,Article ,Antigenic drift ,Virus ,Madin Darby Canine Kidney Cells ,Microbiology ,03 medical and health sciences ,Dogs ,Immune system ,Influenza A Virus, H9N2 Subtype ,Influenza A virus ,medicine ,Animals ,Multidisciplinary ,business.industry ,Vaccination ,Embryonated ,food and beverages ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Poultry farming ,Virology ,Cold Temperature ,030104 developmental biology ,Influenza Vaccines ,Influenza in Birds ,business ,Chickens - Abstract
To prevent H9N2 avian influenza virus infection in chickens, a long-term vaccination program using inactivated vaccines has been implemented in China. However, the protective efficacy of inactivated vaccines against antigenic drift variants is limited, and H9N2 influenza virus continues to circulate in vaccinated chicken flocks in China. Therefore, developing a cross-reactive vaccine to control the impact of H9N2 influenza in the poultry industry remains a high priority. In the present study, we developed a live cold-adapted H9N2 influenza vaccine candidate (SD/01/10-ca) by serial passages in embryonated eggs at successively lower temperatures. A total of 13 amino acid mutations occurred during the cold-adaptation of this H9N2 virus. The candidate was safe in chickens and induced robust hemagglutination-inhibition antibody responses and influenza virus–specific CD4+ and CD8+ T cell immune responses in chickens immunized intranasally. Importantly, the candidate could confer protection of chickens from homologous and heterogenous H9N2 viruses. These results demonstrated that the cold-adapted attenuated H9N2 virus would be selected as a vaccine to control the infection of prevalent H9N2 influenza viruses in chickens.
- Published
- 2016
21. Transmission and pathogenicity of novel reassortants derived from Eurasian avian-like and 2009 pandemic H1N1 influenza viruses in mice and guinea pigs
- Author
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Huijie Gao, Jinhua Liu, Yuelong Shu, Yu Wang, Juan Pu, Kin-Chow Chang, Lirong Liu, Qiming He, Hanchun Yang, Qinfang Liu, Honglei Sun, Xin Guo, Yipeng Sun, Weili Kong, Zhihua Qin, and Dayan Wang
- Subjects
0301 basic medicine ,Genes, Viral ,Genotype ,Guinea Pigs ,Respiratory System ,Sus scrofa ,030106 microbiology ,Reassortment ,Virulence ,Hemagglutinin Glycoproteins, Influenza Virus ,Receptors, Cell Surface ,Biology ,Virus Replication ,medicine.disease_cause ,Article ,Virus ,Madin Darby Canine Kidney Cells ,Birds ,Guinea pig ,03 medical and health sciences ,Dogs ,Influenza A Virus, H1N1 Subtype ,Influenza, Human ,Reassortant Viruses ,Influenza A virus ,medicine ,Animals ,Humans ,Pandemics ,Mice, Inbred BALB C ,Multidisciplinary ,Coinfection ,Virology ,HEK293 Cells ,030104 developmental biology ,Viral replication ,Influenza Vaccines ,Influenza in Birds ,Hybridization, Genetic ,Female ,Protein Binding - Abstract
Given the present extensive co-circulation in pigs of Eurasian avian-like (EA) swine H1N1 and 2009 pandemic (pdm/09) H1N1 viruses, reassortment between them is highly plausible but largely uncharacterized. Here, experimentally co-infected pigs with a representative EA virus and a pdm/09 virus yielded 55 novel reassortant viruses that could be categorized into 17 genotypes from Gt1 to Gt17 based on segment segregation. Majority of novel reassortants were isolated from the lower respiratory tract. Most of reassortant viruses were more pathogenic and contagious than the parental EA viruses in mice and guinea pigs. The most transmissible reassortant genotypes demonstrated in guinea pigs (Gt2, Gt3, Gt7, Gt10 and Gt13) were also the most lethal in mice. Notably, nearly all these highly virulent reassortants (all except Gt13) were characterized with possession of EA H1 and full complement of pdm/09 ribonucleoprotein genes. Compositionally, we demonstrated that EA H1-222G contributed to virulence by its ability to bind avian-type sialic acid receptors, and that pdm/09 RNP conferred the most robust polymerase activity to reassortants. The present study revealed high reassortment compatibility between EA and pdm/09 viruses in pigs, which could give rise to progeny reassortant viruses with enhanced virulence and transmissibility in mice and guinea pig models.
- Published
- 2016
22. Truncation of C-terminal 20 amino acids in PA-X contributes to adaptation of swine influenza virus in pigs
- Author
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Yu Wang, Xin Xiong, Honglei Sun, Hanchun Yang, Xuxiao Zhang, Ming Jiang, Juan Pu, Qiming He, Jinhua Liu, Yipeng Sun, Qinfang Liu, Xin Guo, and Guanlong Xu
- Subjects
0301 basic medicine ,Swine ,viruses ,Viral Nonstructural Proteins ,Biology ,Real-Time Polymerase Chain Reaction ,medicine.disease_cause ,Article ,Virus ,Cell Line ,Madin Darby Canine Kidney Cells ,law.invention ,03 medical and health sciences ,Dogs ,Orthomyxoviridae Infections ,Genes, Reporter ,law ,Influenza A Virus, H1N2 Subtype ,Influenza A virus ,medicine ,Animals ,Amino Acids ,Lung ,Gene ,Swine Diseases ,chemistry.chemical_classification ,Translational frameshift ,Multidisciplinary ,Virology ,Fusion protein ,Recombinant Proteins ,Amino acid ,Repressor Proteins ,Trachea ,030104 developmental biology ,Microscopy, Fluorescence ,chemistry ,Viral replication ,Interferon Type I ,Recombinant DNA ,Nasal Cavity - Abstract
The PA-X protein is a fusion protein incorporating the N-terminal 191 amino acids of the PA protein with a short C-terminal sequence encoded by an overlapping ORF (X-ORF) in segment 3 that is accessed by + 1 ribosomal frameshifting and this X-ORF exists in either full length or a truncated form (either 61-or 41-condons). Genetic evolution analysis indicates that all swine influenza viruses (SIVs) possessed full-length PA-X prior to 1985, but since then SIVs with truncated PA-X have gradually increased and become dominant, implying that truncation of this protein may contribute to the adaptation of influenza virus in pigs. To verify this hypothesis, we constructed PA-X extended viruses in the background of a “triple-reassortment” H1N2 SIV with truncated PA-X and evaluated their biological characteristics in vitro and in vivo. Compared with full-length PA-X, SIV with truncated PA-X had increased viral replication in porcine cells and swine respiratory tissues, along with enhanced pathogenicity, replication and transmissibility in pigs. Furthermore, we found that truncation of PA-X improved the inhibition of IFN-I mRNA expression. Hereby, our results imply that truncation of PA-X may contribute to the adaptation of SIV in pigs.
- Published
- 2016
23. A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses
- Author
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Qingdong Tang, Bao Jingnan, Honglei Sun, Juan Pu, Yipeng Sun, Jinliang Wang, and Jinhua Liu
- Subjects
China ,animal structures ,Newcastle Disease ,animal diseases ,Population ,Newcastle disease virus ,Hemagglutinin (influenza) ,medicine.disease_cause ,Sensitivity and Specificity ,Newcastle disease ,law.invention ,law ,Virology ,Genotype ,medicine ,Animals ,Multiplex ,education ,Poultry Diseases ,Polymerase chain reaction ,DNA Primers ,education.field_of_study ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,virus diseases ,biology.organism_classification ,Influenza A virus subtype H5N1 ,Real-time polymerase chain reaction ,Influenza A virus ,Influenza in Birds ,biology.protein ,Chickens ,Multiplex Polymerase Chain Reaction - Abstract
Avian influenza viruses (AIVs) and Newcastle disease viruses (NDVs) co-circulate in the poultry population in China. These viruses cause repeated disease outbreaks that exhibit similar clinical symptoms and epidemiological patterns. H5 and H9 influenza viruses are the major pathogens infecting poultry stocks. Recently, H3 AIV (one of the main subtypes in waterfowl) has become endemic in chickens. A multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was designed for simultaneous detection and differentiation of avian H3, H5, H9 subtype AIVs and NDVs. Four primer sets were evaluated, three of which specifically targeted the hemagglutinin genes of H3, H5 and H9 AIVs, while the other targeted the NDV fusion gene. The sensitivity and specificity of the mRT-PCR assay was determined. The assay detected the major clades or genotypes of all of the reference AIVs and NDVs currently circulating in China. In addition, the mRT-PCR results obtained from screening 380 clinical swabs and 12 experimental tracheal samples were consistent with those obtained using conventional virus isolation methods. The mRT-PCR assay was established successfully for the detection and differentiation of avian H3, H5, and H9 subtype AIVs and NDVs. The method should, therefore, provide a valuable diagnostic tool for these infections.
- Published
- 2012
24. Natural and experimental infection of dogs with pandemic H1N1/2009 influenza virus
- Author
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Shasha Sun, Yipeng Sun, Jingyi Zhao, Degui Lin, Linghong Fan, Honglei Sun, Jingjiao Ma, Lijie Du, Juan Pu, and Jinhua Liu
- Subjects
China ,viruses ,Virus isolation ,Molecular Sequence Data ,Biology ,medicine.disease_cause ,Virus ,Dogs ,Influenza A Virus, H1N1 Subtype ,Orthomyxoviridae Infections ,Virology ,Influenza, Human ,Pandemic ,Influenza A virus ,medicine ,Animals ,Humans ,Dog Diseases ,Pandemics ,Phylogeny ,Inoculation ,Transmission (medicine) ,virus diseases ,Pathogenicity ,Female - Abstract
Evidence of H1N1/2009 influenza virus infection was identified in two domestic dogs in China in November 2009. Virus isolation and sequence analysis of all eight genes of the two isolates showed that they were related closely to the H1N1/2009 influenza virus circulating in humans, indicating that they were probably acquired from humans. To determine the pathogenicity and transmissibility of H1N1/2009 influenza virus in dogs, experimental infection and transmission were performed. Inoculated dogs were able to shed virus in nasal secretions, but symptoms were very mild. Uninoculated dogs were co-mingled to determine the transmissibility of the isolate, and one of three exposed dogs was shown to develop infection. The present findings indicate that human H1N1/2009 can infect dogs, but is transmitted inefficiently between dogs.
- Published
- 2012
25. Twenty amino acids at the C-terminus of PA-X are associated with increased influenza A virus replication and pathogenicity
- Author
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Jiao Hu, Qiming He, Lu Qi, Xin Xiong, Yang Lin, Kin-Chow Chang, Jinliang Wang, Yu Wang, Yipeng Sun, Honglei Sun, Xiufan Liu, Huijie Gao, Juan Pu, Weili Kong, Lai-Giea Seng, and Jinhua Liu
- Subjects
viruses ,Amino Acid Motifs ,Virulence ,Viral Nonstructural Proteins ,Biology ,Virus Replication ,medicine.disease_cause ,H5N1 genetic structure ,Negative-strand RNA Viruses ,Ribosomal frameshift ,Virus ,Mice ,Influenza A Virus, H1N1 Subtype ,Virology ,Influenza, Human ,Influenza A Virus, H9N2 Subtype ,medicine ,Influenza A virus ,Animals ,Humans ,Mice, Inbred BALB C ,Influenza A Virus, H5N1 Subtype ,Animal ,Standard ,Influenza A virus subtype H5N1 ,Reverse genetics ,Repressor Proteins ,Viral replication ,Female - Abstract
The PA-X protein, arising from ribosomal frameshift during PA translation, was recently discovered in influenza A virus (IAV). The C-terminal domain ‘X’ of PA-X proteins in IAVs can be classified as full-length (61 aa) or truncated (41 aa). In the main, avian influenza viruses express full-length PA-X proteins, whilst 2009 pandemic H1N1 (pH1N1) influenza viruses harbour truncated PA proteins. The truncated form lacks aa 232–252 of the full-length PA-X protein. The significance of PA-X length in virus function remains unclear. To address this issue, we constructed a set of contemporary influenza viruses (pH1N1, avian H5N1 and H9N2) with full and truncated PA-X by reverse genetics to compare their replication and host pathogenicity. All full-length PA-X viruses in human A549 cells conferred 10- to 100-fold increase in viral replication and 5–8 % increase in apoptosis relative to corresponding truncated PA-X viruses. Full-length PA-X viruses were more virulent and caused more severe inflammatory responses in mice. Furthermore, aa 233–252 at the C terminus of PA-X strongly suppressed co-transfected gene expression by ∼50 %, suggesting that these terminal 20 aa could play a role in enhancing viral replication and contribute to virulence.
- Published
- 2015
26. Characterization of clade 2.3.4.4 highly pathogenic H5 avian influenza viruses in ducks and chickens
- Author
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Xiufan Liu, Juan Pu, Litao Liu, Guanlong Xu, Jiao Hu, George F. Gao, Jinhua Liu, and Honglei Sun
- Subjects
0301 basic medicine ,Gene Expression Regulation, Viral ,Antigenicity ,viruses ,Highly pathogenic ,Virulence ,Hemagglutinin Glycoproteins, Influenza Virus ,Biology ,medicine.disease_cause ,Microbiology ,03 medical and health sciences ,Species Specificity ,medicine ,Influenza A virus ,Animals ,Clade ,Antigens, Viral ,General Veterinary ,business.industry ,Transmission (medicine) ,virus diseases ,General Medicine ,Poultry farming ,Virology ,Influenza A virus subtype H5N1 ,030104 developmental biology ,Ducks ,Influenza Vaccines ,Influenza in Birds ,business ,Chickens - Abstract
Worldwide dissemination of reassortant variants of H5 clade 2.3.4.4 highly pathogenic avian influenza (HPAI) viruses has posed a great threat to the poultry industry. Here, we systematically characterized the H5N2, H5N6 and H5N8 influenza viruses in poultry and compared them with those of previous clade 2.3.4 H5N1 virus. All the three H5 subtype reassortants caused systematic infection in ducks, and exhibited efficient direct transmission in ducks. All of them were highly pathogenic in chickens; however, the H5 reassortants have reduced virulence compared to the parental H5N1 virus. Antigenicity analysis revealed that the current vaccines that are widely used in China may fail to confer protection against the H5 reassortants.
- Published
- 2015
27. Serological survey of canine H3N2, pandemic H1N1/09, and human seasonal H3N2 influenza viruses in cats in northern China, 2010–2014
- Author
-
Bo Jiang, Degui Lin, Ming Wang, Juan Pu, Lijie Du, Xuxiao Zhang, Ran Wang, Honglei Sun, Jinhua Liu, Ye Shen, and Yipeng Sun
- Subjects
Male ,Veterinary medicine ,medicine.medical_specialty ,China ,viruses ,Prevalence ,Short Report ,Biology ,medicine.disease_cause ,Antibodies, Viral ,Cat Diseases ,Antibodies ,Dogs ,Influenza A Virus, H1N1 Subtype ,Orthomyxoviridae Infections ,Seroepidemiologic Studies ,Virology ,Pandemic ,Influenza, Human ,Influenza A virus ,medicine ,Seroprevalence ,Animals ,Humans ,Dog Diseases ,CATS ,Transmission (medicine) ,Public health ,Influenza A Virus, H3N2 Subtype ,virus diseases ,Cat ,Influenza ,respiratory tract diseases ,Infectious Diseases ,Human mortality from H5N1 ,Cats ,Female ,Seasons - Abstract
Background The close contact between cats and humans poses a threat to public health because of the potential zoonotic transmission of influenza viruses to humans. Therefore, we examined the seroprevalence of pandemic H1N1/09, canine H3N2, and human H3N2 viruses in pet cats in northern China from 2010 to 2014. Finding Of 1794 serum samples, the seropositivity rates for H1N1/09, canine H3N2, and human H3N2 were 5.7%, 0.7%, and 0.4%, respectively. The seropositivity rate for H1N1/09 in cats was highest in 2010 (8.3%), and then declined continuously thereafter. Cats older than 10 years were most commonly seropositive for the H1N1/09 virus. Conclusions Our findings emphasize the need for continuous surveillance of influenza viruses in cats in China.
- Published
- 2015
28. The infection of turkeys and chickens by reassortants derived from pandemic H1N1 2009 and avian H9N2 influenza viruses
- Author
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Jinhua Liu, Litao Liu, Sizhe Wu, Yu Wang, Yipeng Sun, Juan Pu, Yi Qu, Honglei Sun, Chong Li, Ye Shen, Yu Zhou, and Weili Kong
- Subjects
Turkeys ,animal structures ,viruses ,Reassortment ,Virulence ,Biology ,medicine.disease_cause ,Virus Replication ,Virus ,Article ,Disease Outbreaks ,Viral Proteins ,Influenza A Virus, H1N1 Subtype ,Ribonucleases ,Reassortant Viruses ,medicine ,Influenza A virus ,Influenza A Virus, H9N2 Subtype ,Animals ,Infectivity ,Multidisciplinary ,Coinfection ,virus diseases ,Virology ,Influenza A virus subtype H5N1 ,Enzyme Activation ,Kinetics ,Viral replication ,Influenza in Birds ,Chickens - Abstract
Outbreaks of pandemic H1N1 2009 (pH1N1) in turkeys have been reported in several countries. Co-infection of pH1N1 and avian H9N2 influenza viruses in turkeys provide the opportunity for their reassortment and novel reassortant viruses might further be transmitted to other avian species. However, virulence and transmission of those reassortant viruses in poultry remain unclear. In the present study, we generated 16 single-gene reassortant influenza viruses including eight reassortants on the pH1N1 background by individual replacement with a corresponding gene segment from H9N2 and eight reassortants on the H9N2 background replaced individually with corresponding gene from pH1N1 and characterized reassortants viruses in turkeys and chickens. We found that the pH1N1 virus dramatically increased its infectivity and transmissibility in turkeys and chickens after introducing any gene (except for PB2) from H9N2 virus and H9N2 virus acquired single gene (except for HA) of pH1N1 almost did not influence its replication and transmission in turkeys and chickens. Additionally, 13 reassortant viruses transmitted from turkeys to chickens. Our results indicate that turkeys and chickens are susceptible to pH1N1-H9N2 reassortant viruses and mixing breeding of different avian species would facilitate the transmission of these reassortant viruses.
- Published
- 2014
29. C-terminal elongation of NS1 of H9N2 influenza virus induces a high level of inflammatory cytokines and increases transmission
- Author
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Yu Wang, Qiming He, Honglei Sun, Lirong Liu, Zhihua Qin, Yipeng Sun, Sizhe Wu, Rui Zhang, Weili Kong, Jinhua Liu, Jinliang Wang, and Juan Pu
- Subjects
China ,viruses ,Mutant ,Biology ,Viral Nonstructural Proteins ,medicine.disease_cause ,Virus Replication ,Virus ,Proinflammatory cytokine ,Cell Line ,Dogs ,Orthomyxoviridae Infections ,Virology ,Influenza A virus ,medicine ,Influenza A Virus, H9N2 Subtype ,Animals ,Mice, Inbred BALB C ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Influenza A virus subtype H5N1 ,Reverse genetics ,Reverse Genetics ,Disease Models, Animal ,Viral replication ,Cell culture ,Influenza in Birds ,Host-Pathogen Interactions ,Cytokines ,Female ,Chickens - Abstract
H9N2 avian influenza viruses are enzootic around the world and can infect many different avian and mammalian hosts, including humans. Unlike the H9N2 viruses, which mainly originated in other countries and possess a non-structural protein 1 (NS1) of 230 aa, 98 % of the H9N2 viruses isolated in China lack the 13 aa at the C terminus of NS1 (217 aa in total). The biological significance of NS1 elongation remains elusive. To examine the effect of NS1 C-terminal elongation in the influenza virus, we used reverse genetics to generate a wt avian influenza H9N2 virus containing a 217 aa NS1 (H9N2NS1217) and two mutant viruses with elongated NS1s of 230 and 237 aa (H9N2NS1230 and H9N2NS1237). C-terminal elongation of NS1 did not have a significant impact on virus replication in Madin–Darby canine kidney cells or DF-1 cells. The three variants exhibited similar replicability in mice; however, the H9N2NS1230 and H9N2NS1237 variants exhibited an upregulation in the level of inflammatory cytokines. In addition, both the H9N2NS1230 and H9N2NS1237 viruses increased replication and induced a high level of inflammatory cytokines and transmission in chickens, compared with the wt virus. These findings suggest that the NS1 extension conferred a gain of fitness to some extent.
- Published
- 2014
30. Influenza A Virus Acquires Enhanced Pathogenicity and Transmissibility after Serial Passages in Swine
- Author
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Kai Wei, Juan Pu, Zhenhong Sun, Kin-Chow Chang, Yanbo Yin, Xin Guo, Guangpeng Ma, Weili Kong, Jinhua Liu, Hanchun Yang, Yipeng Sun, and Honglei Sun
- Subjects
Swine ,viruses ,Immunology ,Guinea Pigs ,Viral quasispecies ,Biology ,Recombinant virus ,medicine.disease_cause ,Microbiology ,Airborne transmission ,H5N1 genetic structure ,Virus ,Cell Line ,Dogs ,Orthomyxoviridae Infections ,Serial passage ,Virology ,Influenza A virus ,medicine ,Animals ,Serial Passage ,Virulence ,Viral culture ,Insect Science ,Mutation ,Pathogenesis and Immunity ,Female ,Bronchoalveolar Lavage Fluid - Abstract
Genetic and phylogenetic analyses suggest that the pandemic H1N1/2009 virus was derived from well-established swine influenza lineages; however, there is no convincing evidence that the pandemic virus was generated from a direct precursor in pigs. Furthermore, the evolutionary dynamics of influenza virus in pigs have not been well documented. Here, we subjected a recombinant virus (rH1N1) with the same constellation makeup as the pandemic H1N1/2009 virus to nine serial passages in pigs. The severity of infection sequentially increased with each passage. Deep sequencing of viral quasispecies from the ninth passage found five consensus amino acid mutations: PB1 A469T, PA 1129T, NA N329D, NS1 N205K, and NEP T48N. Mutations in the hemagglutinin (HA) protein, however, differed greatly between the upper and lower respiratory tracts. Three representative viral clones with the five consensus mutations were selected for functional evaluation. Relative to the parental virus, the three viral clones showed enhanced replication and polymerase activity in vitro and enhanced replication, pathogenicity, and transmissibility in pigs, guinea pigs, and ferrets in vivo . Specifically, two mutants of rH1N1 (PB1 A469T and a combination of NS1 N205K and NEP T48N) were identified as determinants of transmissibility in guinea pigs. Crucially, one mutant viral clone with the five consensus mutations, which also carried D187E, K211E, and S289N mutations in its HA, additionally was able to infect ferrets by airborne transmission as effectively as the pandemic virus. Our findings demonstrate that influenza virus can acquire viral characteristics that are similar to those of the pandemic virus after limited serial passages in pigs. IMPORTANCE We demonstrate here that an engineered reassortant swine influenza virus, with the same gene constellation pattern as the pandemic H1N1/2009 virus and subjected to only nine serial passages in pigs, acquired greatly enhanced virulence and transmissibility. In particular, one representative pathogenic passaged virus clone, which carried three mutations in the HA gene and five consensus mutations in PB1, PA, NA, NS1, and NEP genes, additionally was able to confer respiratory droplet transmission as effectively as the pandemic H1N1/2009 virus. Our findings suggest that pigs can readily induce adaptive mutational changes to a precursor pandemic-like virus to transform it into a highly virulent and infectious form akin to that of the pandemic H1N1/2009 virus, which underlines the potential direct role of pigs in promoting influenza A virus pathogenicity and transmissibility.
- Published
- 2014
31. Structural basis for preferential avian receptor binding by the human-infecting H10N8 avian influenza virus
- Author
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Wei Zhang, Chaobin Huang, Jianxun Qi, Yuelong Shu, Fei Wang, Xiangdong Li, Jianfang Zhou, Ying Wu, Honglei Sun, Yi Shi, Min Wang, Jinghua Yan, Yuhai Bi, George F. Gao, and Jinhua Liu
- Subjects
China ,Viral protein ,Molecular Sequence Data ,General Physics and Astronomy ,Hemagglutinin Glycoproteins, Influenza Virus ,Plasma protein binding ,Biology ,medicine.disease_cause ,Crystallography, X-Ray ,Influenza A Virus, H7N9 Subtype ,H5N1 genetic structure ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Protein Structure, Secondary ,Protein structure ,Influenza A Virus, H10N8 Subtype ,Influenza, Human ,Influenza A virus ,medicine ,Animals ,Humans ,Receptor ,Multidisciplinary ,General Chemistry ,Surface Plasmon Resonance ,Virology ,Influenza A virus subtype H5N1 ,Recombinant Proteins ,Protein Structure, Tertiary ,Intestines ,Trachea ,Ducks ,Influenza in Birds ,Receptors, Virus ,Protein Binding - Abstract
Since December 2013, at least three cases of human infections with H10N8 avian influenza virus have been reported in China, two of them being fatal. To investigate the epidemic potential of H10N8 viruses, we examined the receptor binding property of the first human isolate, A/Jiangxi-Donghu/346/2013 (JD-H10N8), and determined the structures of its haemagglutinin (HA) in complex with both avian and human receptor analogues. Our results suggest that JD-H10N8 preferentially binds the avian receptor and that residue R137-localized within the receptor-binding site of HA-plays a key role in this preferential binding. Compared with the H7N9 avian influenza viruses, JD-H10N8 did not exhibit the enhanced binding to human receptors observed with the prevalent H7N9 virus isolate Anhui-1, but resembled the receptor binding activity of the early-outbreak H7N9 isolate (Shanghai-1). We conclude that the H10N8 virus is a typical avian influenza virus.
- Published
- 2014
32. Comparative virus replication and host innate responses in human cells infected with three prevalent clades (2.3.4, 2.3.2, and 7) of highly pathogenic avian influenza H5N1 viruses
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Honglei Sun, Yi Zhang, Jing Li, Qingyu Zhu, Kin-Chow Chang, Jinhua Liu, Juan Pu, Yipeng Sun, and Jiang Gu
- Subjects
Influenza A Virus, H5N1 Subtype ,Host (biology) ,viruses ,Immunology ,Biology ,medicine.disease_cause ,Virus Replication ,Microbiology ,Virology ,H5N1 genetic structure ,Influenza A virus subtype H5N1 ,Virus ,Immunity, Innate ,Viral replication ,Immunity ,Insect Science ,medicine ,Influenza A virus ,Pathogenesis and Immunity ,Humans ,Clade - Abstract
Highly pathogenic avian influenza H5N1 virus clades 2.3.4, 2.3.2, and 7 are the dominant cocirculating H5N1 viruses in poultry in China. However, humans appear to be clinically susceptible mostly to the 2.3.4 virus clade. Here, we demonstrated that A549 cells and human macrophages infected with clade 2.3.4 viruses produced significantly more viruses than those infected with the other two clades. Likewise, clade 2.3.4-infected macrophages caused the most severe cellular damage and strongest proinflammatory response.
- Published
- 2014
33. A Single Amino Acid at the Hemagglutinin Cleavage Site Contributes to the Pathogenicity and Neurovirulence of H5N1 Influenza Virus in Mice
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Yipeng Sun, Honglei Sun, Yi Shi, Yi Zhang, Qingyu Zhu, Jing Li, Xishan Lu, Yuhai Bi, Hanchun Yang, George F. Gao, Juan Pu, and Jinhua Liu
- Subjects
Immunology ,Amino Acid Motifs ,Molecular Sequence Data ,Virulence ,Hemagglutinin Glycoproteins, Influenza Virus ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Cell Line ,Serine ,Mice ,Virology ,Influenza, Human ,Influenza A virus ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Peptide sequence ,Tropism ,Genetics ,Mice, Inbred BALB C ,Influenza A Virus, H5N1 Subtype ,Reverse genetics ,Influenza A virus subtype H5N1 ,Insect Science ,Influenza in Birds ,Pathogenesis and Immunity ,Female ,Chickens ,Protein Processing, Post-Translational - Abstract
H5 influenza viruses containing a motif of multiple basic amino acids at the hemagglutinin (HA) cleavage site (HACS) are highly pathogenic in chicken but display different virulence phenotypes in mammals. Previous studies have shown that multiple basic amino acids of H5N1 influenza virus are a prerequisite for lethality in mice. However, it remains unclear which specific residue at the cleavage site affects the pathogenicity of H5N1 in mammals. A comprehensive genetic analysis of the H5N1 HACS showed that residues at P6 (position 325, by H3 numbering) were the most polymorphic, including serine (S), arginine (R), deletion (*), glycine (G), and isoleucine (I). To determine whether a single residue at P6 could affect virulence, we introduced different mutations at P6 of an avirulent clade 7 H5N1 strain, rg325G, by reverse genetics. Among the recombinant viruses, the rg325S virus showed the highest cleavage efficiency in vitro . All these viruses were highly pathogenic in chicken but exhibited different virulences in mice. The rg325S virus exhibited the highest pathogenicity in terms of unrestricted organ tropism and neurovirulence. Remarkably, the HA-325S substitution dramatically increased the pathogenicity of H5N1 viruses of other clades, including clades 2.2, 2.3.2, and 2.3.4, indicating that this residue impacts genetically divergent H5N1 viruses. An analysis of predicted structures containing these mutations showed that the cleavage site loop with 325S was the most exposed, which might be responsible for the efficient cleavage and high virulence. Our results demonstrate that an amino acid substitution at the P6 cleavage site alone could modulate the virulence of H5N1 in mice.
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- 2012
34. Acute respiratory distress syndrome induced by a swine 2009 H1N1 variant in mice
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Lihong Fan, Jian Qiao, Yuan Ma, Jinhua Liu, Guangpeng Ma, Juan Pu, Yi Zhang, Jun Yang, Yipeng Sun, and Honglei Sun
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ARDS ,Pathology ,Viral Diseases ,Anatomy and Physiology ,Pulmonology ,Swine ,viruses ,Respiratory System ,lcsh:Medicine ,Hemagglutinin Glycoproteins, Influenza Virus ,medicine.disease_cause ,Virus Replication ,Leukocyte Count ,Mice ,Influenza A Virus, H1N1 Subtype ,Edema ,Influenza A virus ,Diffuse alveolar damage ,lcsh:Science ,Lung ,Respiratory Distress Syndrome ,Multidisciplinary ,Animal Models ,medicine.anatomical_structure ,Infectious Diseases ,Cytokines ,Medicine ,Female ,medicine.symptom ,Bronchoalveolar Lavage Fluid ,Research Article ,medicine.medical_specialty ,Lung injury ,Microbiology ,Virus ,Proinflammatory cytokine ,Model Organisms ,Virology ,medicine ,Animals ,Humans ,Biology ,business.industry ,lcsh:R ,medicine.disease ,respiratory tract diseases ,Disease Models, Animal ,Kinetics ,Immunology ,Mutation ,lcsh:Q ,Blood Gas Analysis ,business - Abstract
Background Acute respiratory distress syndrome (ARDS) induced by pandemic 2009 H1N1 influenza virus has been widely reported and was considered the main cause of death in critically ill patients with 2009 H1N1 infection. However, no animal model has been developed for ARDS caused by infection with 2009 H1N1 virus. Here, we present a mouse model of ARDS induced by 2009 H1N1 virus. Methodology Principal Findings Mice were inoculated with A/swine/Shandong/731/2009 (SD/09), which was a 2009 H1N1 influenza variant with a G222D mutation in the hemagglutinin. Clinical symptoms were recorded every day. Lung injury was assessed by lung water content and histopathological observation. Arterial blood gas, leukocyte count in the bronchial alveolar lavage fluid and blood, virus titers, and cytokine levels in the lung were measured at various times post-inoculation. Mice infected with SD/09 virus showed typical ARDS symptoms characterized by 60% lethality on days 8–10 post-inoculation, highly edematous lungs, inflammatory cellular infiltration, alveolar and interstitial edema, lung hemorrhage, progressive and severe hypoxemia, and elevated levels of proinflammatory cytokines and chemokines. Conclusions/Significance These results suggested that we successfully established an ARDS mouse model induced by a virulent 2009 H1N1 variant without previous adaptation, which may be of benefit for evaluating the pathogenesis or therapy of human ARDS caused by 2009 H1N1 virus.
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- 2011
35. M Gene Reassortment in H9N2 Influenza Virus Promotes Early Infection and Replication: Contribution to Rising Virus Prevalence in Chickens in China.
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Juan Pu, Honglei Sun, Yi Qu, Chenxi Wang, Weihua Gao, Junda Zhu, Yipeng Sun, Yuhai Bi, Yinhua Huang, Kin-Chow Chang, Jie Cui, and Jinhua Liu
- Subjects
- *
INFLUENZA treatment , *INFLUENZA A virus , *VIRAL replication , *DISEASE prevalence , *MUTAGENESIS , *CHICKEN diseases - Abstract
Segment reassortment and base mutagenesis of influenza A viruses are the primary routes to the rapid evolution of high-fitness virus genotypes. We recently described a predominant G57 genotype of avian H9N2 viruses that caused countrywide outbreaks in chickens in China during 2010 to 2013, which led to the zoonotic emergence of H7N9 viruses. One of the key features of the G57 genotype is the replacement of the earlier A/chicken/Beijing/1/1994 (BJ/94)-like M gene with the A/quail/Hong Kong/G1/1997 (G1)-like M gene of quail origin. We report here the functional significance of the G1-like M gene in H9N2 viruses in conferring increased infection severity and infectivity in primary chicken embryonic fibroblasts and chickens. H9N2 virus housing the G1-like M gene, in place of the BJ/94-like M gene, showed an early surge in viral mRNA and viral RNA (vRNA) transcription that was associated with enhanced viral protein production and with an early elevated release of progeny virus comprising largely spherical rather than filamentous virions. Importantly, H9N2 virus with the G1-like M gene conferred extrapulmonary virus spread in chickens. Five highly represented signature amino acid residues (37A, 95K, 224N, and 242N in the M1 protein and 21G in the M2 protein) encoded by the prevalent G1-like M gene were demonstrated to be prime contributors to enhanced infectivity. Therefore, the genetic evolution of the M gene in H9N2 virus increases reproductive virus fitness, indicating its contribution to the rising virus prevalence in chickens in China. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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36. Serological survey of canine H3N2, pandemic H1N1/09, and human seasonal H3N2 influenza viruses in cats in northern China, 2010-2014.
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Xuxiao Zhang, Ye Shen, Lijie Du, Ran Wang, Bo Jiang, Honglei Sun, Juan Pu, Degui Lin, Ming Wang, Jinhua Liu, and Yipeng Sun
- Subjects
INFLUENZA A virus, H3N2 subtype ,CAT diseases ,SEROPREVALENCE ,INFLUENZA A virus ,DISEASE prevalence ,IMMUNOGLOBULINS - Abstract
Background: The close contact between cats and humans poses a threat to public health because of the potential zoonotic transmission of influenza viruses to humans. Therefore, we examined the seroprevalence of pandemic H1N1/09, canine H3N2, and human H3N2 viruses in pet cats in northern China from 2010 to 2014. Finding: Of 1794 serum samples, the seropositivity rates for H1N1/09, canine H3N2, and human H3N2 were 5.7%, 0.7%, and 0.4%, respectively. The seropositivity rate for H1N1/09 in cats was highest in 2010 (8.3%), and then declined continuously thereafter. Cats older than 10 years were most commonly seropositive for the H1N1/09 virus. Conclusions: Our findings emphasize the need for continuous surveillance of influenza viruses in cats in China. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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37. Recombinant parainfluenza virus 5 expressing clade 2.3.4.4b H5 hemagglutinin protein confers broad protection against H5Ny influenza viruses.
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Han Li, Haoran Sun, Mengyan Tao, Qiqi Han, Haili Yu, Jiaqi Li, Xue Lu, Qi Tong, Juan Pu, Yipeng Sun, Litao Liu, Jinhua Liu, and Honglei Sun
- Subjects
- *
INFLUENZA A virus , *PARAINFLUENZA viruses , *AVIAN influenza A virus , *RECOMBINANT viruses , *INFLUENZA viruses , *HEMAGGLUTININ , *PANDEMIC preparedness - Abstract
The global circulation of clade 2.3.4.4b H5Ny highly pathogenic avian influenza viruses (HPAIVs) in poultry and wild birds, increasing mammal infections, continues to pose a public health threat and may even form a pandemic. An efficacious vaccine against H5Ny HPAIVs is crucial for emergency use and pandemic preparedness. In this study, we developed a parainfluenza virus 5 (PIV5)-based vaccine candidate expressing hemagglutinin (HA) protein of clade 2.3.4.4b H5 HPAIV, termed rPIV5-H5, and evaluated its safety and efficacy in mice and ferrets. Our results demonstrated that intranasal immunization with a single dose of rPIV5-H5 could stimulate H5-specific antibody responses, moreover, a prime-boost regimen using rPIV5-H5 stimulated robust humoral, cellular, and mucosal immune responses in mice. Challenge study showed that rPIV5-H5 prime-boost regimen provided sterile immunity against lethal clade 2.3.4.4b H5N1 virus infection in mice and ferrets. Notably, rPIV5-H5 prime-boost regimen provided protection in mice against challenge with lethal doses of heterologous clades 2.2, 2.3.2, and 2.3.4 H5N1, and clade 2.3.4.4h H5N6 viruses. These results revealed that rPIV5-H5 can elicit protective immunity against a diverse clade of highly pathogenic H5Ny virus infection in mammals, highlighting the potential of rPIV5-H5 as a pan-H5 influenza vaccine candidate for emergency use. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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- View/download PDF
38. Identification of swine influenza A virus and Stenotrophomonas maltophilia co-infection in Chinese pigs
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Jinhua Liu, Yuhai Bi, Jun Yang, Guanghua Fu, Honglei Sun, Dongjun Hou, Yipeng Sun, and Juan Pu
- Subjects
China ,Swine ,Stenotrophomonas maltophilia ,viruses ,Guinea Pigs ,Population ,Microbial Sensitivity Tests ,Biology ,medicine.disease_cause ,S. maltophilia ,H5N1 genetic structure ,Virus ,lcsh:Infectious and parasitic diseases ,Microbiology ,Swine influenza virus ,Orthomyxoviridae Infections ,Virology ,Drug Resistance, Bacterial ,Influenza A virus ,medicine ,Animals ,lcsh:RC109-216 ,Viral shedding ,education ,Swine Diseases ,education.field_of_study ,Co-infections ,Coinfection ,Research ,virus diseases ,Viral Load ,medicine.disease ,biology.organism_classification ,Anti-Bacterial Agents ,Virus Shedding ,Trachea ,Nasal Mucosa ,Infectious Diseases ,Female ,Gram-Negative Bacterial Infections ,Viral load - Abstract
Background Influenza virus virulence can be exacerbated by bacterial co-infections. Swine influenza virus (SIV) infection together with some bacteria is found to enhance pathogenicity. Methods SIV-positive samples suspected of containing bacteria were used for bacterial isolation and identification. Antimicrobial susceptibility testing was performed by disc diffusion methods. To investigate the interaction of SIV and the bacteria in vitro, guinea pigs were used as mammalian hosts to determine the effect on viral susceptibility and transmissibility. Differences in viral titers between groups were compared using Student’s t-test. Results During surveillance for SIV in China from 2006 to 2009, seven isolates (24.14%) of 29 influenza A viruses were co-isolated with Stenotrophomonas maltophilia from nasal and tracheal swab samples of pigs. Antimicrobial susceptibility testing showed that the bacteria possessed a high level of resistance towards clinically used antibiotics. To investigate the interaction between these two microorganisms in influencing viral susceptibility and transmission in humans, guinea pigs were used as an infection model. Animals were inoculated with SIV or S. maltophilia alone or co-infected with SIV and S. maltophilia. The results showed that although no transmission among guinea pigs was observed, virus–bacteria co-infections resulted in higher virus titers in nasal washes and trachea and a longer virus shedding period. Conclusions This is the first report of influenza virus co-infection with S. maltophilia in the Chinese swine population. Increased replication of virus by co-infection with multidrug resistant bacteria might increase the infection rate of SIV in humans. The control of S. maltophilia in clinics will contribute to reducing the spread of SIV in pigs and humans.
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39. Reassortment with Dominant Chicken H9N2 Influenza Virus Contributed to the Fifth H7N9 Virus Human Epidemic.
- Author
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Juan Pu, Yanbo Yin, Jiyu Liu, Xinyu Wang, Yong Zhou, Zejiang Wang, Yipeng Sun, Honglei Sun, Fangtao Li, Jingwei Song, Runkang Qu, Weihua Gao, Dongdong Wang, Zhen Wang, Shijie Yan, Mingyue Chen, Jinfeng Zeng, Zhimin Jiang, Haoran Sun, and Yanan Zong
- Subjects
- *
INFLUENZA A virus, H7N9 subtype , *INFLUENZA A virus , *INFLUENZA viruses , *CHICKEN diseases , *AVIAN influenza A virus , *VIRUS diseases - Abstract
H9N2 avian influenza virus (AIV) is regarded as a principal donor of viral genes through reassortment to cocirculating influenza viruses that can result in zoonotic reassortants. Whether H9N2 virus can maintain a sustained evolutionary impact on such reassortants is unclear. Since 2013, avian H7N9 virus had caused five sequential human epidemics in China; the fifth wave in 2016 to 2017 was by far the largest, but the mechanistic explanation behind the scale of infection is not clear. Here, we found that just prior to the fifth H7N9 virus epidemic, H9N2 viruses had phylogenetically mutated into new subclades, changed antigenicity, and increased their prevalence in chickens vaccinated with existing H9N2 vaccines. In turn, the new H9N2 virus subclades of PB2 and PA genes, housing mammalian adaptive mutations, were reassorted into cocirculating H7N9 virus to create a novel dominant H7N9 virus genotype that was responsible for the fifth H7N9 virus epidemic. H9N2-derived PB2 and PA genes in H7N9 virus conferred enhanced polymerase activity in human cells at 33°C and 37°C and increased viral replication in the upper and lower respiratory tracts of infected mice, which could account for the sharp increase in human cases of H7N9 virus infection in the 2016-2017 epidemic. The role of H9N2 virus in the continual mutation of H7N9 virus highlights the public health significance of H9N2 virus in the generation of variant reassortants of increasing zoonotic potential. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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40. Truncation of PA-X contributes to virulence and transmission of H3N8 and H3N2 canine influenza viruses in dogs.
- Author
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Litao Liu, Shikai Song, Ye Shen, Chao Ma, Tong Wang, Qi Tong, Honglei Sun, Juan Pu, Iqbal, Munir, Jinhua Liu, and Yipeng Sun
- Subjects
- *
INFLUENZA A virus , *INFLUENZA viruses , *AVIAN influenza A virus , *GENE expression profiling , *DOGS , *RECOMBINANT viruses , *PLASMIDS - Abstract
Equine-origin H3N8 and avian-origin H3N2 canine influenza viruses (CIVs) prevalent in dogs are thought to pose a public health threat arising from intimate contact between dogs and humans. However, our understanding of CIV virulence is still limited. Influenza A virus PA-X is a fusion protein encoded in part by a +1 frameshifted open reading frame (X-ORF) in segment 3. The X-ORF can be translated in full-length (61 amino acids) or truncated (41 amino acids) form. Genetic analysis indicated that the X-ORFs of equine H3N8 and avian H3N2 influenza viruses encoded 61 amino acids but were truncated after introduction into dogs. To determine the effect of PA-X truncation on the biological characteristics of CIVs, we constructed four recombinant viruses on H3N8 and H3N2 CIV backgrounds bearing truncated or full-length PA-Xs. We observed that truncation of PA-X increased growth of both H3N8 and H3N2 CIVs in MDCK cells and suppressed expression from co-transfected plasmids in MDCK cells. Furthermore, truncation of PA-X enhanced viral pathogenicity in dogs as shown by aggravated clinical symptoms and histopathological changes, increased viral replication in the respiratory system, and prolonged virus shedding. Additionally, CIVs with truncated PA-Xs were transmitted more efficiently in dogs. Global gene expression profiling of the lungs of infected dogs revealed that differentially expressed genes were mainly associated with inflammatory responses, which might contribute to the pathogenicity of PA-X-truncated CIVs. Our findings revealed that truncation of PA-X might be important for the adaptation of influenza viruses to dogs. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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41. An R195K Mutation in the PA-X Protein Increases the Virulence and Transmission of Influenza A Virus in Mammalian Hosts.
- Author
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Yipeng Sun, Zhe Hu, Xuxiao Zhang, Mingyue Chen, Zhen Wang, Guanlong Xu, Yuhai Bi, Qi Tong, Mingyang Wang, Honglei Sun, Juan Pu, Iqbal, Munir, and Jinhua Liu
- Subjects
- *
H1N1 influenza , *INFLUENZA A virus , *INFLUENZA A virus, H7N9 subtype , *AVIAN influenza A virus , *REVERSE genetics , *FERRET - Abstract
In the 21st century, the emergence of H7N9 and H1N1/2009 influenza viruses, originating from animals and causing severe human infections, has prompted investigations into the genetic alterations required for cross-species transmission. We previously found that replacement of the human-origin PA gene segment in avian influenza virus (AIV) could overcome barriers to cross-species transmission. Recently, it was reported that the PA gene segment encodes both the PA protein and a second protein, PA-X. Here, we investigated the role of PA-X. We found that an H9N2 avian influenza reassortant virus bearing a human-origin H1N1/2009 PA gene was attenuated in mice after the loss of PA-X. Reverse genetics analyses of PA-X substitutions conserved in human influenza viruses indicated that R195K, K206R, and P210L substitutions conferred significantly increased replication and pathogenicity on H9N2 virus in mice and ferrets. PA-X R195K was present in all human H7N9 and H1N1/2009 viruses and predominated in human H5N6 viruses. Compared with PA-X 195R, H7N9 influenza viruses bearing PA-X 195K showed increased replication and transmission in ferrets. We further showed that PA-X 195K enhanced lung inflammatory responses, potentially due to decreased host shutoff function. A competitive transmission study in ferrets indicated that 195K provides a replicative advantage over 195R in H1N1/2009 viruses. In contrast, PA-X 195K did not influence the virulence of H9N2 AIV in chickens, suggesting that the effects of the substitution were mammal specific. Therefore, future surveillance efforts should scrutinize this region of PA-X because of its potential impact on cross-species transmission of influenza viruses. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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42. Amino Acid 316 of Hemagglutinin and the Neuraminidase Stalk Length Influence Virulence of H9N2 Influenza Virus in Chickens and Mice.
- Author
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Yipeng Sun, Yuanyuan Tan, Kai Wei, Honglei Sun, Yi Shi, Juan Pu, Hanchun Yang, Gao, George F., Yanbo Yin, Wenhai Feng, Perez, Daniel R., and Jinhua Liu
- Subjects
- *
INFLUENZA A virus , *HEMAGGLUTININ , *NEURAMINIDASE , *ERYTHROCYTES , *ENZYMES - Abstract
H9N2 influenza viruses with an A316S substitution in hemagglutinin (HA) and a shorter neuraminidase (NA) stalk have become predominant in China. The A316S was shown to increase HA cleavage efficiency when combined with short stalk NA, and the short stalk NA improved NA enzyme activity and release of virus from erythrocytes. Single mutations or combinations of these mutations strengthened the virulence of H9N2 virus in chickens and mice. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
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