Monoclonal antibodies(mAbs) are widely used in virus research and disease diagnosis. The nucleoprotein(NP) of influenza A virus(IAV) plays important roles in multiple stages of the virus life cycle. Therefore, generat...Monoclonal antibodies(mAbs) are widely used in virus research and disease diagnosis. The nucleoprotein(NP) of influenza A virus(IAV) plays important roles in multiple stages of the virus life cycle. Therefore, generating conserved mAbs against NP and characterizing their properties will provide useful tools for IAV research. In this study, two mAbs against the NP protein, 10 E9 and 3 F3, were generated with recombinant truncated NP proteins(NP-1 and NP-2) as immunogens. The heavy-chain subclass of both 10 E9 and 3 F3 was determined to be IgG2α, and the light-chain type was κ. Truncation and site-specific mutation analyses showed that the epitopes of mAbs 10 E9 and 3 F3 were located in the N terminal 84–89 amino acids and the C terminal 320–324 amino acids of the NP protein, respectively. We found that mAbs 10 E9 and 3 F3 reacted well with the NP protein of H1–H15 subtypes of IAV. Both 10 E9 and 3 F3 can be used in immunoprecipitation assay, and 10 E9 was also successfully applied in confocal microscopy. Furthermore, we found that the 10 E9-recognized _(84) SAGKDP_(89) epitope and 3 F3-recognized 320 ENPAH324 epitope were highly conserved in NP among all avian and human IAVs. Thus, the two mAbs we developed could be used as powerful tools in the development of diagnostic methods of IAV, and also surely promote the basic research in understanding the replication mechanisms of IAV.展开更多
Purpose:In the present study,we focused on the 46 microRNAs and 719 genes in the microRNA-gene network,reported by us,and aimed to build a research blueprint of feedforward loops and reveal the key TFs in H1N1-infecte...Purpose:In the present study,we focused on the 46 microRNAs and 719 genes in the microRNA-gene network,reported by us,and aimed to build a research blueprint of feedforward loops and reveal the key TFs in H1N1-infected mouse lung.Method:Based on microRNAs and genes in the microRNA-gene network previously reported by us,we used Jemboss software to find relationships between TFs and microRNAs(or genes),and then built a TF-microRNA-gene network exploiting the interactions between TFs and microRNAs(or genes).Next,we searched the sequences of above genes or microRNAs near the transcription start site(TSS)area,and then used the MatchTM algorithm to predict relevant TFs,and built the TF-Gene-Network.Result:We built a TF-microRNAgene network and exploreed eight key TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin,in the network,and then constructed subgraphs of these eight TFs.Simultaneously,we predicted the possible target genes of microRNAs and identified the feedforward regulation relationship of possible TFs,microRNAs and mRNAs.The results showed that all eight factors with a score greater than 100 were TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,CRX,myogenin and MZF1.We then constructed subtables of the above eight TFs.Conclusion:In this study,TFs including NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin showed the highest score(>100)not only in the TF-microRNA-gene network but also in feedforward loops,indicating that these eight TFs play the most important roles in mouse H1N1 influenza virus infection biology.展开更多
Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple...Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple online databases were used to search and screen out the active components from MHT,the related targets of active components of MHT and the genes related to IAV.Search the corresponding genes name of target through UniProt database.Cytoscape 3.7.2 was used to construct the drug-component-target network diagram.Venn diagram was used to screen the intersection genes of the active components corresponding to the target and disease-related genes,and the intersection genes were imported into the STRING Database Online platform to obtain the protein-protein interaction(PPI)network.Then,the PPI network was imported into Cytoscape 3.7.2 to obtain the core targets.Finally,Wei Sheng Xing(http://www.bioinformatics.com.cn/)was used to do GO function enrichment analysis and KEGG signaling pathway enrichment analysis for the intersection genes,and the results of GO and KEGG were visualized.Results A total of 116 active components and 253 potential targets were screened from MHT.Quercetin,Kaempferol,and Luteolin are the main active components,and AKT1,TNF,TP53,IL6,and JUN are the core targets of 253 potential targets.There are 2906 targets of influenza A,including 121 intersection genes.Enrichment analysis showed that there were 131 entries of molecular function,89 entries of cell component,1645 entries of biological process and 195 entries of signaling pathway.PI3K-Akt,MAPK,and JAK-STAT were the main signaling pathways.Conclusion MHT plays an important role in the prevention and treatment of influenza A by acting on multiple targets and multiple signaling pathways.展开更多
Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membran...Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membranes. In this study, we identified cation-dependent mannose-6-phosphate receptor(M6PR) as a crucial host factor for the replication of IAV. We found that siRNA knockdown of M6PR expression significantly reduced the growth titers of different subtypes of IAV, and that the inhibitory effect of M6PR siRNA treatment on IAV growth was overcome by the complement of exogenously expressed M6PR. When A549 cells were treated with siRNA targeting M6PR,the nuclear accumulation of viral nucleoprotein(NP) was dramatically inhibited at early timepoints post-infection, indicating that M6PR engages in the early stage of the IAV replication cycle. By investigating the role of M6PR in the individual entry and post-entry steps of IAV replication, we found that the downregulation of M6PR expression had no effect on attachment, internalization, early endosome trafficking,or late endosome acidification. However, we found that M6PR expression was critical for the fusion of viral envelope and late endosomal membranes. Of note, M6PR interacted with the hemagglutinin(HA) protein of IAV, and further studies showed that the lumenal domain of M6PR and the ectodomain of HA2 mediated the interaction and directly promoted the fusion of the viral and late endosomal membranes,thereby facilitating IAV replication. Together, our findings highlight the importance of the M6PR–HA interaction in the fusion of viral and late endosomal membranes during IAV replication.展开更多
Background:The influenza A virus is the primary cause of respiratory infections and poses a global health risk.Pudilan Xiaoyan oral liquid(PDL)exhibits anti-inflammatory and immunomodulatory properties.PDL is commonly...Background:The influenza A virus is the primary cause of respiratory infections and poses a global health risk.Pudilan Xiaoyan oral liquid(PDL)exhibits anti-inflammatory and immunomodulatory properties.PDL is commonly employed in clinical practice to manage upper respiratory tract infections.However,there is still much to uncover regarding its potential therapeutic mechanism.Methods:Institute of cancer research mice were infected with influenza A virus via nasal drip.The general state of the mice,lung index,and lung index inhibition rate were used to evaluate the efficacy of PDL.Enzyme-linked immunosorbent assay,western blotting,and immunohistochemistry were used to observe the presence of proteins and cytokines in the lung tissue.Apoptosis was evaluated using the TUNEL assay.Results:PDL improved the mental state of influenza A virus-infected mice,reduced the lung index,and inhibited viral replication.The expression of interleukin-1βand tumor necrosis factor-αwere decreased,whereas the expression of interleukin-10 in the lung tissue was increased due to PDL treatment.In addition,PDL treatment modulated Toll-like receptor 4 and MyD88 expressions in the lung tissues.PDL significantly reduced apoptosis and decreased cleaved caspase-3 and PARP levels,whereas increased B-cell lymphoma-2 expression in the lung tissue.Notably,the moderate-dose group of PDL exhibited a more pronounced effect.These findings indicate that PDL exerts a protective effect against pneumonia injury in influenza A virus-infected mice.Conclusion:PDL inhibited the inflammatory response and regulated apoptosis by regulating Toll-like receptor 4 and MyD88 protein expressions,thereby protecting the lung tissue from viral infection-induced lung tissue injury.展开更多
Influenza A virus(IAV)genome comprises eight negative-sense RNA segments,of which the replication is well orchestrated and the delicate balance of multiple segments are dynamically regulated throughout IAV life cycle....Influenza A virus(IAV)genome comprises eight negative-sense RNA segments,of which the replication is well orchestrated and the delicate balance of multiple segments are dynamically regulated throughout IAV life cycle.However,previous studies seldom discuss these balances except for functional hemagglutinin-neuraminidase balance that is pivotal for both virus entry and release.Therefore,we attempt to revisit IAV life cycle by highlighting the critical role of“genome balance”.Moreover,we raise a“balance regression”model of IAV evolution that the virus evolves to rebalance its genome after reassortment or interspecies transmission,and direct a“balance compensation”strategy to rectify the“genome imbalance”as a result of artificial modifications during creation of recombinant IAVs.This review not only improves our understanding of IAV life cycle,but also facilitates both basic and applied research of IAV in future.展开更多
Influenza A virus(IAV),responsible for seasonal epidemics and recurring pandemics,represents a global threat to public health.Given the risk of a potential IAV pandemic,it is increasingly important to better understan...Influenza A virus(IAV),responsible for seasonal epidemics and recurring pandemics,represents a global threat to public health.Given the risk of a potential IAV pandemic,it is increasingly important to better understand virushost interactions and develop new anti-viral strategies.Here,we reported nonmuscle myosin IIA(MYH9)-mediated regulation of IAV infection.MYH9 depletion caused a profound inhibition of IAV infection by reducing viral attachment and internalization in human lung epithelial cells.Surprisingly,overexpression of MYH9 also led to a significant reduction in viral productive infection.Interestingly,overexpression of MYH9 retained viral attachment,internalization,or uncoating,but suppressed the viral ribonucleoprotein(vRNP)activity in a minigenome system.Further analyses found that excess MYH9 might interrupt the formation of vRNP by interacting with the viral nucleoprotein(NP)and result in the reduction of the completed vRNP in the nucleus,thereby inhibiting subsequent viral RNA transcription and replication.Together,we discovered that MYH9 can interact with IAV NP protein and engage in the regulation of vRNP complexes,thereby involving viral replication.These findings enlighten new mechanistic insights into the complicated interface of host-IAV interactions,ultimately making it an attractive target for the generation of antiviral drugs.展开更多
Influenza viruses not only cause respiratory illness,but also have been reported to elicit neurological manifestations following acute viral infection.The central nervous system(CNS)has a specific defense mechanism ag...Influenza viruses not only cause respiratory illness,but also have been reported to elicit neurological manifestations following acute viral infection.The central nervous system(CNS)has a specific defense mechanism against pathogens structured by cerebral microvasculature lined with brain endothelial cells to form the blood–brain barrier(BBB).To investigate the response of human brain microvascular endothelial cells(hBMECs)to the Influenza A virus(IAV),we inoculated the cells with the A/WSN/33(H1N1)virus.We then conducted an RNAseq experiment to determine the changes in gene expression levels and the activated disease pathways following infection.The analysis revealed an effective activation of the innate immune defense by inducing the pattern recognition receptors(PRRs).Along with the production of proinflammatory cytokines,we detected an upregulation of interferons and interferon-stimulated genes,such as IFN-β/λ,ISG15,CXCL11,CXCL3 and IL-6,etc.Moreover,infected hBMECs exhibited a disruption in the cytoskeletal structure both on the transcriptomic and cytological levels.The RNAseq analysis showed different pathways and candidate genes associated with the neuroactive ligand-receptor interaction,neuroinflammation,and neurodegenerative diseases,together with a predicted activation of the neuroglia.Likewise,some genes linked with the mitochondrial structure and function displayed a significantly altered expression.En masse,this data supports that hBMECs could be infected by the IAV,which induces the innate and inflammatory immune response.The results suggest that the influenza virus infection could potentially induce a subsequent aggravation of neurological disorders.展开更多
Honeysuckle has been used in the treatment of influenza virus infection for thousands of years in China.However,its main active components and the functional mechanisms remain to be elucidated.Here,four honeysuckle ex...Honeysuckle has been used in the treatment of influenza virus infection for thousands of years in China.However,its main active components and the functional mechanisms remain to be elucidated.Here,four honeysuckle extracts,including acids extract,flavonoids extract,total extract and acids-flavonoids mixture,were prepared to clarify the main active antiviral components.The cytopathic effect reduction assay showed that all the four extracts inhibited the replication of influenza viruses H1N1,H3N2 and the oseltamivir-resistant mutant strain H1N1-H275Y.The acids-flavonoids mixture had the strongest inhibitory effects in vitro with EC_(50) values of 3.8,4.1,and >20 lg/m L against H1N1,H3N2 and H1N1-H275Y,respectively,showing competitive antiviral activity with oseltamivir and ribavirin.Honeysuckle acids extract also showed the most significant antiviral activity in vivo.Oral administration of the acids extract at a dosage of 600 mg/kg/d effectively alleviated viral pneumonia,maintained body weight and improved the survival rate to 30% of the mice infected with a lethal dose of H1N1.The results of time-of-drug addition experiment and neuraminidase (NA) inhibition assay showed that honeysuckle extracts had a broad-spectrum inhibitory effect against influenza virus NAs.The flavonoid extract showed the strongest inhibitory effect on the NA of influenza virus H7N9 with an IC_(50) of 24.7 lg/m L.These results suggested that these extracts might exert their antiviral activity by suppressing the release of influenza viruses.Briefly,our findings demonstrate that acids and flavonoids extracts of honeysuckle are the major antiviral active components,and the acids extract has the potential to be developed into an antiviral agent against influenza virus,especially for oseltamivirresistant viruses.展开更多
OBJECTIVE:To investigate synergistic effect of Reduning(RDN)injection plus ribavirin against severe pneumonia induced by H1 N1 influenza A virus in mice.METHODS:We established a mouse model of severe pneumonia induced...OBJECTIVE:To investigate synergistic effect of Reduning(RDN)injection plus ribavirin against severe pneumonia induced by H1 N1 influenza A virus in mice.METHODS:We established a mouse model of severe pneumonia induced by influenza A virus by infecting Balb/c mice with CA07 virus.We randomly assigned the infected mice into four groups,and treated them with normal saline(NS group),RDN(injection,86.6 mg/kg),ribavirin(injection,66.6 mg/kg)or double Ribavirin plus RDN group,the same dosage as used in the single treatments)for 5 d.Lung index and lung pathology were recorded or calculated in terms of the curative effective.Cytokines,NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome related protein including caspase-associated recruitment domain(CARD)domain Apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1 and NOD-like receptor family,pyrin domain containing 3(NLRP3),and reactive oxygen species were simultaneously investigated.RESULTS:RDN plus ribavirin treatment,not RDN or ribavirin alone,provided a significant survival benefit to the influenza A virus-infected mice.The combination treatment protected the mice against severe influenza infection by attenuating the severe lung injury.The combined treatment also reduced the viral titers in mouse lungs and lung index,downregulated their immunocytokine levels,including IL-1βand IL-18,and down regulated the NLRP3,especially the transcription and translation of caspase-1.Meanwhile NS group had significantly higher reactive oxygen species(ROS)expression which could was dramatically reduced by the treatment of RDN plus ribavirin.CONCLUSION:Our study showed that RDN combined with ribavirin could protect the mice,and reduce the lung immunopathologic damage caused by severe influenza pneumonia.The mechanism could be that it reduced ROS produce and inhibited NLRP3 inflammasome activation so that mainly lower the downstream inflammatory cytokines IL-1βand IL-18.展开更多
As a major component of the viral ribonucleoprotein(vRNP)complex in influenza A virus(IAV),nucleoprotein(NP)interacts with isoforms of importinαfamily members,leading to the import of itself and vRNP complex into the...As a major component of the viral ribonucleoprotein(vRNP)complex in influenza A virus(IAV),nucleoprotein(NP)interacts with isoforms of importinαfamily members,leading to the import of itself and vRNP complex into the nucleus,a process pivotal in the replication cycle of IAV.In this study,we found that BinCARD1,an isoform of Bcl10-interacting protein with CARD(BinCARD),was leveraged by IAV for efficient viral replication.BinCARD1 promoted the nuclear import of the vRNP complex and newly synthesized NP and thus enhanced vRNP complex activity.Moreover,we found that BinCARD1 interacted with NP to promote NP binding to importinα7,an adaptor in the host nuclear import pathway.However,we also found that BinCARD1 promoted RIG-I-mediated innate immune signaling by mediating Lys63-linked polyubiquitination of TRAF3,and that TBK1 appeared to degrade BinCARD1.We showed that BinCARD1 was polyubiquitinated at residue K103 through a Lys63 linkage,which was recognized by the TBK1-p62 axis for autophagic degradation.Overall,our data demonstrate that IAV leverages BinCARD1 as an important host factor that promotes viral replication,and two mechanisms in the host defense system are triggered—innate immune signaling and autophagic degradation—to mitigate the promoting effect of BinCARD1 on the life cycle of IAV.展开更多
Seasonal influenza epidemics and influenza pandemics caused by influenza A virus(IAV)has resulted in millions of deaths in the world.The development of anti-IAV vaccines and therapeutics is urgently needed for prevent...Seasonal influenza epidemics and influenza pandemics caused by influenza A virus(IAV)has resulted in millions of deaths in the world.The development of anti-IAV vaccines and therapeutics is urgently needed for prevention and treatment of IAV infection and for controlling future influenza pandemics.Hemagglutinin(HA)of IAV plays a critical role in viral binding,fusion and entry,and contains the major neutralizing epitopes.Therefore,HA is an attractive target for developing anti-IAV drugs and vaccines.Here we have reviewed the recent progress in study of conformational changes of HA during viral fusion process and development of HA-based antiviral therapeutics and vaccines.展开更多
Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus.Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibi...Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus.Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors.In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA,which expresses Gaussia luciferase upon influenza A virus infection.Using this cell line,an assay was developed and optimized to search for inhibitors of influenza virus replication.Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format(Z 0 factor value40.8).A pilot screening provides further evidence for validation of the assay.Taken together,this work provides a simple,convenient,and reliable HTS assay to identify compounds with anti-influenza activity.展开更多
Efficient and eco-friendly disinfection of air-borne human respiratory RNA viruses is pursued in both public environment and portable usage.The AlGaN-based deep ultraviolet(DUV)light-emission diode(LED)has high practi...Efficient and eco-friendly disinfection of air-borne human respiratory RNA viruses is pursued in both public environment and portable usage.The AlGaN-based deep ultraviolet(DUV)light-emission diode(LED)has high practical potentials because of its advantages of variable wavelength,rapid sterilization,environmental protection,and miniaturization.Therefore,whether the emission wavelength has effects on the disinfection as well as whether the device is feasible to sterilize various respiratory RNA viruses under portable conditions is crucial.Here,we fabricate AlGaN-based DUV LEDs with different wavelength on high-temperature-annealed(HTA)AlN/Sapphire templates and investigate the inactivation effects for several respiratory RNA viruses.The AlN/AlGaN superlattices are employed between the template and upper n-AlGaN to release the strong compressive stress(SCS),improving the crystal quality and interface roughness.DUV LEDs with the wavelength of 256,265,and 278 nm,corresponding to the light output power of 6.8,9.6,and 12.5 mW,are realized,among which the 256 nm-LED shows the most potent inactivation effect in human respiratory RNA viruses,including SARS-CoV-2,influenza A virus(IAV),and human parainfluenza virus(HPIV),at a similar light power density(LPD)of~0.8 mW/cm2 for 10 s.These results will contribute to the advanced DUV LED application of disinfecting viruses with high potency and broad spectrum in a portable and eco-friendly use.展开更多
Background:Endothelial cells play a key role in the cytokine storm caused by influenza A virus. MicroRNA-155 (miR-155) is an important regulator in inflammation. Its role in the inflammatory response to influenza A in...Background:Endothelial cells play a key role in the cytokine storm caused by influenza A virus. MicroRNA-155 (miR-155) is an important regulator in inflammation. Its role in the inflammatory response to influenza A infection, however, has yet to be elucidated. In this study, we explored the role as well as the underlying mechanism of miR-155 in the cytokine production in influenza A-infected endothelial cells.Methods:Human pulmonary microvascular endothelial cells (HPMECs) were infected with the influenza A virus strain H1N1. The efficiency of H1N1 infection was confirmed by immunofluorescence. The expression levels of proinflammatory cytokines and miR-155 were determined using real-time polymerase chain reaction. A dual-luciferase reporter assay characterized the interaction between miR-155 and sphingosine-1-phosphate receptor 1 (S1PR1). Changes in the target protein levels were determined using Western blot analysis.Results:MiR-155 was elevated in response to the H1N1 infection in HPMECs (24 h post-infection vs. 0 h post-infection, 3.875 ± 0.062 vs. 1.043 ± 0.013, P = 0.001). Over-expression of miR-155 enhanced inflammatory cytokine production (miR-155 mimic vs. negative control, all P < 0.05 in regard of cytokine levels) and activation of nuclear factor kappa B in infected HPMECs (miR-155 mimic vs. negative control, P = 0.004), and down-regulation of miR-155 had the opposite effect. In addition, S1PR1 was a direct target of miR-155 in the HPMECs. Inhibition of miR-155 enhanced the expression of the S1PR1 protein. Down-regulation of S1PR1 decreased the inhibitory effect of the miR-155 blockade on H1N1-induced cytokine production and nuclear factor kappa B activation in HPMECs. Conclusion:MiR-155 maybe modulate influenza A-induced inflammatory response by targeting S1PR1.展开更多
In this study,SB216763 and cyclosporine A were identified as anti-influenza A virus(IAV)agents by tran-scriptome signature reversion(TSR)analysis through deep mining of the cellular transcriptome of hu-man airway and ...In this study,SB216763 and cyclosporine A were identified as anti-influenza A virus(IAV)agents by tran-scriptome signature reversion(TSR)analysis through deep mining of the cellular transcriptome of hu-man airway and lung cell lines infected with 3 strains of IAV and the chemical perturbations library.A synergistic effect of SB216763 and cyclosporine A against influenza A was disclosed by quantification of the network-based relationship,which was validated in vitro.Along with burgeoning omics approaches,transcriptome-based drug development is flourishing,which provides a novel insight into antivirals dis-covery with comprehensive cellular transcriptional information of disease and chemical perturbations in multicomponent intervention.This strategy can be applied as a new approach in discovering multitar-get antiviral agents from approved drugs,clinical compounds,natural products or other known bioactive compounds.展开更多
Mutations in viral proteins can lead to the cold adaption of influenza A virus and the cold-adapted virus is an important vaccination instrument.Here,we identify a novel strain of influenza A virus with cold sensitivi...Mutations in viral proteins can lead to the cold adaption of influenza A virus and the cold-adapted virus is an important vaccination instrument.Here,we identify a novel strain of influenza A virus with cold sensitivity conferred by a mutation at a phosphorylation site within the nucleoprotein(NP).The highly conserved tyrosine 385 residue(Y385)of NP was identified as a phosphorylation site by mass spectrometry.The constructive NP phosphorylation mimicked by Y385 E mutation was fatal for virus replication,while the continuous Y385 dephosphorylation mimicked by Y385 F mutation had little impact on virus replication in vitro.Notably,the Y385 F virus showed much lower replicative capacity in turbinates of mice compared with the wild type virus.Moreover,the replication of Y385 F virus was significantly reduced in both A549 and MDCK cells grown at 33℃,when compared to that at 37℃.These results indicated that the Y385 F mutation led to cold sensitivity of virus.We further found that the cold sensitivity of Y385 F virus could be attributed to diminished NP oligomerization rather than any changes in intracellular localization.Taken together,these findings suggest that the phosphorylation of NP may be a critical factor that regulates the temperature sensitivity of influenza A virus.展开更多
Modern miniaturization and the digitalization of characterization instruments greatly facilitate the diffusion of technological advances in new fields and generate innovative applications.The concept of a portable,ine...Modern miniaturization and the digitalization of characterization instruments greatly facilitate the diffusion of technological advances in new fields and generate innovative applications.The concept of a portable,inexpensive and semi-automated biosensing platform,or lab-on-a-chip,is a vision shared by many researchers and venture industries.Under this scope,we present a semiconductor monolithic integration approach to conduct surface plasmon resonance studies.This technology is already commonly used for biochemical characterization in pharmaceutical industries,but we have reduced the technological platform to a few nanometers in scale on a semiconductor chip.We evaluate the signal quality of this nanophotonic device using hyperspectral-imaging technology,and we compare its performance with that of a standard prism-based commercial system.Two standard biochemical agents are employed for this characterization study:bovine serum albumin and inactivated influenza A virus.Time resolutions of data acquisition varying between 360 and 2.2 s are presented,yielding 2.731025–1.531026 RIU resolutions,respectively.展开更多
A number of eukaryotic expression vectors have been developed for use as DNA vaccines. They showed varying abilities to initiate immune responses;however, there is little data to indicate which of these vectors will b...A number of eukaryotic expression vectors have been developed for use as DNA vaccines. They showed varying abilities to initiate immune responses;however, there is little data to indicate which of these vectors will be the most useful and practical for DNA vaccines in different species. This report examines the use of five expression vectors with different promoters and Kozak sequence to express the same hemagglutinin (HA) protein of an H6N2 avian influenza virus for DNA vaccination in chickens. Although intramuscular vaccination with seven DNA constructs elicited no or limited measurable H6 HA antibody responses in Hy-Line chickens, variable reduction in virus shedding for either oropharyngeal or cloacal swabs post-virus challenge were observed. This indicated that all DNA constructs generated some levels of protective immunity against homologous virus challenge. Interestingly, lower dose (50 or 100 μg) of plasmid DNAs consistently induced better immune response than higher dose (300 or 500 μg). In the transfection experiments there appeared to be a hierarchy in the in vitro expression efficiency in the order of pCAG-optiHAk/ pCAG-HAk > pCI-HAk > VR-HA > pCI-HA > pCI-neo-HA > pVAX-HA. Since the level of in vitro expression correlates with the level of immune response in vivo, in vitro expression levels of the DNA constructs can be used as an indicator for pre-selection of plasmid vaccines prior to in vivo assessment. Moreover, our results suggested that the Kozak sequence could be used as an effective tool for DNA vaccine design.展开更多
Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 1...Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).展开更多
基金supported by the Natural Science Foundation of Heilongjiang Province,China(JQ2019C005)the National Natural Science Foundation of China(31702265 and 32172847)。
文摘Monoclonal antibodies(mAbs) are widely used in virus research and disease diagnosis. The nucleoprotein(NP) of influenza A virus(IAV) plays important roles in multiple stages of the virus life cycle. Therefore, generating conserved mAbs against NP and characterizing their properties will provide useful tools for IAV research. In this study, two mAbs against the NP protein, 10 E9 and 3 F3, were generated with recombinant truncated NP proteins(NP-1 and NP-2) as immunogens. The heavy-chain subclass of both 10 E9 and 3 F3 was determined to be IgG2α, and the light-chain type was κ. Truncation and site-specific mutation analyses showed that the epitopes of mAbs 10 E9 and 3 F3 were located in the N terminal 84–89 amino acids and the C terminal 320–324 amino acids of the NP protein, respectively. We found that mAbs 10 E9 and 3 F3 reacted well with the NP protein of H1–H15 subtypes of IAV. Both 10 E9 and 3 F3 can be used in immunoprecipitation assay, and 10 E9 was also successfully applied in confocal microscopy. Furthermore, we found that the 10 E9-recognized _(84) SAGKDP_(89) epitope and 3 F3-recognized 320 ENPAH324 epitope were highly conserved in NP among all avian and human IAVs. Thus, the two mAbs we developed could be used as powerful tools in the development of diagnostic methods of IAV, and also surely promote the basic research in understanding the replication mechanisms of IAV.
基金National Natural Science Foundation of China(No.81873072)the China Academy of Chinese Medical Sciences Foundation(No.ZZ11-093,No.ZXKT17037).
文摘Purpose:In the present study,we focused on the 46 microRNAs and 719 genes in the microRNA-gene network,reported by us,and aimed to build a research blueprint of feedforward loops and reveal the key TFs in H1N1-infected mouse lung.Method:Based on microRNAs and genes in the microRNA-gene network previously reported by us,we used Jemboss software to find relationships between TFs and microRNAs(or genes),and then built a TF-microRNA-gene network exploiting the interactions between TFs and microRNAs(or genes).Next,we searched the sequences of above genes or microRNAs near the transcription start site(TSS)area,and then used the MatchTM algorithm to predict relevant TFs,and built the TF-Gene-Network.Result:We built a TF-microRNAgene network and exploreed eight key TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin,in the network,and then constructed subgraphs of these eight TFs.Simultaneously,we predicted the possible target genes of microRNAs and identified the feedforward regulation relationship of possible TFs,microRNAs and mRNAs.The results showed that all eight factors with a score greater than 100 were TFs,namely NF-AT1,GKLF,SRY,SOX10,AML1,CRX,myogenin and MZF1.We then constructed subtables of the above eight TFs.Conclusion:In this study,TFs including NF-AT1,GKLF,SRY,SOX10,AML1,MZF1,CRX and myogenin showed the highest score(>100)not only in the TF-microRNA-gene network but also in feedforward loops,indicating that these eight TFs play the most important roles in mouse H1N1 influenza virus infection biology.
基金This study was supported by the Discipline Innovation Team Construction Project of the Second Affiliated Hospital of Shaanxi University of Chinese Medicine (2020XKTD-A02).
文摘Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple online databases were used to search and screen out the active components from MHT,the related targets of active components of MHT and the genes related to IAV.Search the corresponding genes name of target through UniProt database.Cytoscape 3.7.2 was used to construct the drug-component-target network diagram.Venn diagram was used to screen the intersection genes of the active components corresponding to the target and disease-related genes,and the intersection genes were imported into the STRING Database Online platform to obtain the protein-protein interaction(PPI)network.Then,the PPI network was imported into Cytoscape 3.7.2 to obtain the core targets.Finally,Wei Sheng Xing(http://www.bioinformatics.com.cn/)was used to do GO function enrichment analysis and KEGG signaling pathway enrichment analysis for the intersection genes,and the results of GO and KEGG were visualized.Results A total of 116 active components and 253 potential targets were screened from MHT.Quercetin,Kaempferol,and Luteolin are the main active components,and AKT1,TNF,TP53,IL6,and JUN are the core targets of 253 potential targets.There are 2906 targets of influenza A,including 121 intersection genes.Enrichment analysis showed that there were 131 entries of molecular function,89 entries of cell component,1645 entries of biological process and 195 entries of signaling pathway.PI3K-Akt,MAPK,and JAK-STAT were the main signaling pathways.Conclusion MHT plays an important role in the prevention and treatment of influenza A by acting on multiple targets and multiple signaling pathways.
基金supported by the National Natural Science Foundation of China(32192453,32172847)the National Key Research and Development Program of China(2021YFD1800204)+1 种基金the Laboratory of Lingnan Modern Agriculture Project(NT2021007)the earmarked fund for CARS-41。
文摘Influenza A virus(IAV) commandeers numerous host cellular factors for successful replication. However, very few host factors have been revealed to be involved in the fusion of viral envelope and late endosomal membranes. In this study, we identified cation-dependent mannose-6-phosphate receptor(M6PR) as a crucial host factor for the replication of IAV. We found that siRNA knockdown of M6PR expression significantly reduced the growth titers of different subtypes of IAV, and that the inhibitory effect of M6PR siRNA treatment on IAV growth was overcome by the complement of exogenously expressed M6PR. When A549 cells were treated with siRNA targeting M6PR,the nuclear accumulation of viral nucleoprotein(NP) was dramatically inhibited at early timepoints post-infection, indicating that M6PR engages in the early stage of the IAV replication cycle. By investigating the role of M6PR in the individual entry and post-entry steps of IAV replication, we found that the downregulation of M6PR expression had no effect on attachment, internalization, early endosome trafficking,or late endosome acidification. However, we found that M6PR expression was critical for the fusion of viral envelope and late endosomal membranes. Of note, M6PR interacted with the hemagglutinin(HA) protein of IAV, and further studies showed that the lumenal domain of M6PR and the ectodomain of HA2 mediated the interaction and directly promoted the fusion of the viral and late endosomal membranes,thereby facilitating IAV replication. Together, our findings highlight the importance of the M6PR–HA interaction in the fusion of viral and late endosomal membranes during IAV replication.
基金funded by Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences,grant number CI2021A04608National Natural Science Foundation of China,grant number 82141206.
文摘Background:The influenza A virus is the primary cause of respiratory infections and poses a global health risk.Pudilan Xiaoyan oral liquid(PDL)exhibits anti-inflammatory and immunomodulatory properties.PDL is commonly employed in clinical practice to manage upper respiratory tract infections.However,there is still much to uncover regarding its potential therapeutic mechanism.Methods:Institute of cancer research mice were infected with influenza A virus via nasal drip.The general state of the mice,lung index,and lung index inhibition rate were used to evaluate the efficacy of PDL.Enzyme-linked immunosorbent assay,western blotting,and immunohistochemistry were used to observe the presence of proteins and cytokines in the lung tissue.Apoptosis was evaluated using the TUNEL assay.Results:PDL improved the mental state of influenza A virus-infected mice,reduced the lung index,and inhibited viral replication.The expression of interleukin-1βand tumor necrosis factor-αwere decreased,whereas the expression of interleukin-10 in the lung tissue was increased due to PDL treatment.In addition,PDL treatment modulated Toll-like receptor 4 and MyD88 expressions in the lung tissues.PDL significantly reduced apoptosis and decreased cleaved caspase-3 and PARP levels,whereas increased B-cell lymphoma-2 expression in the lung tissue.Notably,the moderate-dose group of PDL exhibited a more pronounced effect.These findings indicate that PDL exerts a protective effect against pneumonia injury in influenza A virus-infected mice.Conclusion:PDL inhibited the inflammatory response and regulated apoptosis by regulating Toll-like receptor 4 and MyD88 protein expressions,thereby protecting the lung tissue from viral infection-induced lung tissue injury.
基金supported by National Natural Science Foundation of China(No.82104134)Key Technology Research and Development Program of Shandong,China(No.2020CXGC010505)The Social Benefiting Technology Program of Qingdao,China(No.21-1-4-rkjk-15-nsh).
文摘Influenza A virus(IAV)genome comprises eight negative-sense RNA segments,of which the replication is well orchestrated and the delicate balance of multiple segments are dynamically regulated throughout IAV life cycle.However,previous studies seldom discuss these balances except for functional hemagglutinin-neuraminidase balance that is pivotal for both virus entry and release.Therefore,we attempt to revisit IAV life cycle by highlighting the critical role of“genome balance”.Moreover,we raise a“balance regression”model of IAV evolution that the virus evolves to rebalance its genome after reassortment or interspecies transmission,and direct a“balance compensation”strategy to rectify the“genome imbalance”as a result of artificial modifications during creation of recombinant IAVs.This review not only improves our understanding of IAV life cycle,but also facilitates both basic and applied research of IAV in future.
基金supported by the National Natural Science Foundation of China(82071788,81901598,81771704,and 82041015)National Key R&D Program of China(2022YFC2604100).
文摘Influenza A virus(IAV),responsible for seasonal epidemics and recurring pandemics,represents a global threat to public health.Given the risk of a potential IAV pandemic,it is increasingly important to better understand virushost interactions and develop new anti-viral strategies.Here,we reported nonmuscle myosin IIA(MYH9)-mediated regulation of IAV infection.MYH9 depletion caused a profound inhibition of IAV infection by reducing viral attachment and internalization in human lung epithelial cells.Surprisingly,overexpression of MYH9 also led to a significant reduction in viral productive infection.Interestingly,overexpression of MYH9 retained viral attachment,internalization,or uncoating,but suppressed the viral ribonucleoprotein(vRNP)activity in a minigenome system.Further analyses found that excess MYH9 might interrupt the formation of vRNP by interacting with the viral nucleoprotein(NP)and result in the reduction of the completed vRNP in the nucleus,thereby inhibiting subsequent viral RNA transcription and replication.Together,we discovered that MYH9 can interact with IAV NP protein and engage in the regulation of vRNP complexes,thereby involving viral replication.These findings enlighten new mechanistic insights into the complicated interface of host-IAV interactions,ultimately making it an attractive target for the generation of antiviral drugs.
基金the financial support provided by the National Program on Key Research Project of China(2016YFD0500406)the National Natural Sciences Foundation of China(Grant No.31872455)+1 种基金the Fundamental Research Funds for the Central Universities(2662018PY016)the Start-up Research Fund from Huazhong Agricultural University.
文摘Influenza viruses not only cause respiratory illness,but also have been reported to elicit neurological manifestations following acute viral infection.The central nervous system(CNS)has a specific defense mechanism against pathogens structured by cerebral microvasculature lined with brain endothelial cells to form the blood–brain barrier(BBB).To investigate the response of human brain microvascular endothelial cells(hBMECs)to the Influenza A virus(IAV),we inoculated the cells with the A/WSN/33(H1N1)virus.We then conducted an RNAseq experiment to determine the changes in gene expression levels and the activated disease pathways following infection.The analysis revealed an effective activation of the innate immune defense by inducing the pattern recognition receptors(PRRs).Along with the production of proinflammatory cytokines,we detected an upregulation of interferons and interferon-stimulated genes,such as IFN-β/λ,ISG15,CXCL11,CXCL3 and IL-6,etc.Moreover,infected hBMECs exhibited a disruption in the cytoskeletal structure both on the transcriptomic and cytological levels.The RNAseq analysis showed different pathways and candidate genes associated with the neuroactive ligand-receptor interaction,neuroinflammation,and neurodegenerative diseases,together with a predicted activation of the neuroglia.Likewise,some genes linked with the mitochondrial structure and function displayed a significantly altered expression.En masse,this data supports that hBMECs could be infected by the IAV,which induces the innate and inflammatory immune response.The results suggest that the influenza virus infection could potentially induce a subsequent aggravation of neurological disorders.
基金supported by the Fundamental Research Funds for the Central Universities (2632017PT03)the National Natural Science Foundation (No. 81700756)。
文摘Honeysuckle has been used in the treatment of influenza virus infection for thousands of years in China.However,its main active components and the functional mechanisms remain to be elucidated.Here,four honeysuckle extracts,including acids extract,flavonoids extract,total extract and acids-flavonoids mixture,were prepared to clarify the main active antiviral components.The cytopathic effect reduction assay showed that all the four extracts inhibited the replication of influenza viruses H1N1,H3N2 and the oseltamivir-resistant mutant strain H1N1-H275Y.The acids-flavonoids mixture had the strongest inhibitory effects in vitro with EC_(50) values of 3.8,4.1,and >20 lg/m L against H1N1,H3N2 and H1N1-H275Y,respectively,showing competitive antiviral activity with oseltamivir and ribavirin.Honeysuckle acids extract also showed the most significant antiviral activity in vivo.Oral administration of the acids extract at a dosage of 600 mg/kg/d effectively alleviated viral pneumonia,maintained body weight and improved the survival rate to 30% of the mice infected with a lethal dose of H1N1.The results of time-of-drug addition experiment and neuraminidase (NA) inhibition assay showed that honeysuckle extracts had a broad-spectrum inhibitory effect against influenza virus NAs.The flavonoid extract showed the strongest inhibitory effect on the NA of influenza virus H7N9 with an IC_(50) of 24.7 lg/m L.These results suggested that these extracts might exert their antiviral activity by suppressing the release of influenza viruses.Briefly,our findings demonstrate that acids and flavonoids extracts of honeysuckle are the major antiviral active components,and the acids extract has the potential to be developed into an antiviral agent against influenza virus,especially for oseltamivirresistant viruses.
基金Supported by The Beijing Natural Science Foundation the research on the mechanisms of Reduning Injection which protects mice from severe pneumonia in terms of the activation level of NLRP3 inflammatomes(No.7172099)
文摘OBJECTIVE:To investigate synergistic effect of Reduning(RDN)injection plus ribavirin against severe pneumonia induced by H1 N1 influenza A virus in mice.METHODS:We established a mouse model of severe pneumonia induced by influenza A virus by infecting Balb/c mice with CA07 virus.We randomly assigned the infected mice into four groups,and treated them with normal saline(NS group),RDN(injection,86.6 mg/kg),ribavirin(injection,66.6 mg/kg)or double Ribavirin plus RDN group,the same dosage as used in the single treatments)for 5 d.Lung index and lung pathology were recorded or calculated in terms of the curative effective.Cytokines,NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome related protein including caspase-associated recruitment domain(CARD)domain Apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1 and NOD-like receptor family,pyrin domain containing 3(NLRP3),and reactive oxygen species were simultaneously investigated.RESULTS:RDN plus ribavirin treatment,not RDN or ribavirin alone,provided a significant survival benefit to the influenza A virus-infected mice.The combination treatment protected the mice against severe influenza infection by attenuating the severe lung injury.The combined treatment also reduced the viral titers in mouse lungs and lung index,downregulated their immunocytokine levels,including IL-1βand IL-18,and down regulated the NLRP3,especially the transcription and translation of caspase-1.Meanwhile NS group had significantly higher reactive oxygen species(ROS)expression which could was dramatically reduced by the treatment of RDN plus ribavirin.CONCLUSION:Our study showed that RDN combined with ribavirin could protect the mice,and reduce the lung immunopathologic damage caused by severe influenza pneumonia.The mechanism could be that it reduced ROS produce and inhibited NLRP3 inflammasome activation so that mainly lower the downstream inflammatory cytokines IL-1βand IL-18.
基金This investigation was funded by a grant from the Laboratory of Lingnan Modern Agriculture Project(NT2021007 to HC and CL)the National Natural Science Foundation of China(NSFC)(32192453 to CL,32172847 to LJ)+2 种基金the National Key Research and Development Program of China(2021YFD1800203 to HC,2021YFD1800204 to CL)the Natural Science Foundation of Heilongjiang Province(JQ2019C005 to CL)the Central Public-Interest Scientific Institution Basal Research Fund(Y2017JC35 to GT).
文摘As a major component of the viral ribonucleoprotein(vRNP)complex in influenza A virus(IAV),nucleoprotein(NP)interacts with isoforms of importinαfamily members,leading to the import of itself and vRNP complex into the nucleus,a process pivotal in the replication cycle of IAV.In this study,we found that BinCARD1,an isoform of Bcl10-interacting protein with CARD(BinCARD),was leveraged by IAV for efficient viral replication.BinCARD1 promoted the nuclear import of the vRNP complex and newly synthesized NP and thus enhanced vRNP complex activity.Moreover,we found that BinCARD1 interacted with NP to promote NP binding to importinα7,an adaptor in the host nuclear import pathway.However,we also found that BinCARD1 promoted RIG-I-mediated innate immune signaling by mediating Lys63-linked polyubiquitination of TRAF3,and that TBK1 appeared to degrade BinCARD1.We showed that BinCARD1 was polyubiquitinated at residue K103 through a Lys63 linkage,which was recognized by the TBK1-p62 axis for autophagic degradation.Overall,our data demonstrate that IAV leverages BinCARD1 as an important host factor that promotes viral replication,and two mechanisms in the host defense system are triggered—innate immune signaling and autophagic degradation—to mitigate the promoting effect of BinCARD1 on the life cycle of IAV.
基金We thank Chungen Pan for assistance in preparing the Fig.4B.SJ and LD were supported by the US NIH grants(Nos.AI68002,AI46221)SL and RL were supported by the Natural Science Foundation of China(Grant No.30772602)the Ministry of Education’s New Century Talent Program(NCET-06-0753).
文摘Seasonal influenza epidemics and influenza pandemics caused by influenza A virus(IAV)has resulted in millions of deaths in the world.The development of anti-IAV vaccines and therapeutics is urgently needed for prevention and treatment of IAV infection and for controlling future influenza pandemics.Hemagglutinin(HA)of IAV plays a critical role in viral binding,fusion and entry,and contains the major neutralizing epitopes.Therefore,HA is an attractive target for developing anti-IAV drugs and vaccines.Here we have reviewed the recent progress in study of conformational changes of HA during viral fusion process and development of HA-based antiviral therapeutics and vaccines.
基金This work was supported in part by National S&T Major Special Project on Major New Drug Innovation(No.2012ZX09301-002-004)(S.C.)National Science and Technology Major Project,“China Mega-Project for Infectious Disease”(No.2013ZX 10004601-002),and the Xiehe Scholar(S.C.).
文摘Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus.Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors.In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA,which expresses Gaussia luciferase upon influenza A virus infection.Using this cell line,an assay was developed and optimized to search for inhibitors of influenza virus replication.Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format(Z 0 factor value40.8).A pilot screening provides further evidence for validation of the assay.Taken together,this work provides a simple,convenient,and reliable HTS assay to identify compounds with anti-influenza activity.
基金supports from the National Key R&D Program of China(2022YFB3605001)National Natural Science Foundation of China(62121005,62004196,61725403,31922004,and 61827813)+2 种基金Youth Innovation Promotion Association of Chinese Academy of Sciences(2023223)Young Elite Scientist Sponsorship Program by CAST(YESS20200182)Innovation Team Project from the Hubei Province(2020CFA015).
文摘Efficient and eco-friendly disinfection of air-borne human respiratory RNA viruses is pursued in both public environment and portable usage.The AlGaN-based deep ultraviolet(DUV)light-emission diode(LED)has high practical potentials because of its advantages of variable wavelength,rapid sterilization,environmental protection,and miniaturization.Therefore,whether the emission wavelength has effects on the disinfection as well as whether the device is feasible to sterilize various respiratory RNA viruses under portable conditions is crucial.Here,we fabricate AlGaN-based DUV LEDs with different wavelength on high-temperature-annealed(HTA)AlN/Sapphire templates and investigate the inactivation effects for several respiratory RNA viruses.The AlN/AlGaN superlattices are employed between the template and upper n-AlGaN to release the strong compressive stress(SCS),improving the crystal quality and interface roughness.DUV LEDs with the wavelength of 256,265,and 278 nm,corresponding to the light output power of 6.8,9.6,and 12.5 mW,are realized,among which the 256 nm-LED shows the most potent inactivation effect in human respiratory RNA viruses,including SARS-CoV-2,influenza A virus(IAV),and human parainfluenza virus(HPIV),at a similar light power density(LPD)of~0.8 mW/cm2 for 10 s.These results will contribute to the advanced DUV LED application of disinfecting viruses with high potency and broad spectrum in a portable and eco-friendly use.
基金a grant from the National Nature Science Foundation of China(No.81470206).
文摘Background:Endothelial cells play a key role in the cytokine storm caused by influenza A virus. MicroRNA-155 (miR-155) is an important regulator in inflammation. Its role in the inflammatory response to influenza A infection, however, has yet to be elucidated. In this study, we explored the role as well as the underlying mechanism of miR-155 in the cytokine production in influenza A-infected endothelial cells.Methods:Human pulmonary microvascular endothelial cells (HPMECs) were infected with the influenza A virus strain H1N1. The efficiency of H1N1 infection was confirmed by immunofluorescence. The expression levels of proinflammatory cytokines and miR-155 were determined using real-time polymerase chain reaction. A dual-luciferase reporter assay characterized the interaction between miR-155 and sphingosine-1-phosphate receptor 1 (S1PR1). Changes in the target protein levels were determined using Western blot analysis.Results:MiR-155 was elevated in response to the H1N1 infection in HPMECs (24 h post-infection vs. 0 h post-infection, 3.875 ± 0.062 vs. 1.043 ± 0.013, P = 0.001). Over-expression of miR-155 enhanced inflammatory cytokine production (miR-155 mimic vs. negative control, all P < 0.05 in regard of cytokine levels) and activation of nuclear factor kappa B in infected HPMECs (miR-155 mimic vs. negative control, P = 0.004), and down-regulation of miR-155 had the opposite effect. In addition, S1PR1 was a direct target of miR-155 in the HPMECs. Inhibition of miR-155 enhanced the expression of the S1PR1 protein. Down-regulation of S1PR1 decreased the inhibitory effect of the miR-155 blockade on H1N1-induced cytokine production and nuclear factor kappa B activation in HPMECs. Conclusion:MiR-155 maybe modulate influenza A-induced inflammatory response by targeting S1PR1.
基金supported by the CAMS Innovation Fund for Medical Sciences (Nos. 2021-I2M-1-028 and 2020I2M-2-010)the Opening Foundation of the State Key Laboratory of Bioactive Substance and Function of Natural Medicines (No. GTZK202109)+2 种基金the National Natural Science Foundation of China (No. 81473256)the Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study (No. BZ0150)the Disciplines Construction Project (No. 201920200802)
文摘In this study,SB216763 and cyclosporine A were identified as anti-influenza A virus(IAV)agents by tran-scriptome signature reversion(TSR)analysis through deep mining of the cellular transcriptome of hu-man airway and lung cell lines infected with 3 strains of IAV and the chemical perturbations library.A synergistic effect of SB216763 and cyclosporine A against influenza A was disclosed by quantification of the network-based relationship,which was validated in vitro.Along with burgeoning omics approaches,transcriptome-based drug development is flourishing,which provides a novel insight into antivirals dis-covery with comprehensive cellular transcriptional information of disease and chemical perturbations in multicomponent intervention.This strategy can be applied as a new approach in discovering multitar-get antiviral agents from approved drugs,clinical compounds,natural products or other known bioactive compounds.
基金supported by grants from the Strategic Priority Research Program of Chinese Academy of Sciences(XDB29010000)the National Natural Science Foundation of China(31630079,31972657,31672531)+2 种基金the National Key Research and Development Program of China(2016YFD0500206)the Mega-Project of Guangxi Natural Science Foundation(2015GXNSFEA139002)supported by Youth Innovation Promotion Association of CAS(2019091)。
文摘Mutations in viral proteins can lead to the cold adaption of influenza A virus and the cold-adapted virus is an important vaccination instrument.Here,we identify a novel strain of influenza A virus with cold sensitivity conferred by a mutation at a phosphorylation site within the nucleoprotein(NP).The highly conserved tyrosine 385 residue(Y385)of NP was identified as a phosphorylation site by mass spectrometry.The constructive NP phosphorylation mimicked by Y385 E mutation was fatal for virus replication,while the continuous Y385 dephosphorylation mimicked by Y385 F mutation had little impact on virus replication in vitro.Notably,the Y385 F virus showed much lower replicative capacity in turbinates of mice compared with the wild type virus.Moreover,the replication of Y385 F virus was significantly reduced in both A549 and MDCK cells grown at 33℃,when compared to that at 37℃.These results indicated that the Y385 F mutation led to cold sensitivity of virus.We further found that the cold sensitivity of Y385 F virus could be attributed to diminished NP oligomerization rather than any changes in intracellular localization.Taken together,these findings suggest that the phosphorylation of NP may be a critical factor that regulates the temperature sensitivity of influenza A virus.
基金The authors acknowledge the financial contribution from the Natural Science and Engineering Research Council of Canada(NSERC Strategic grant STPGP 350501-07)the Canada Research Chair in Quantum Semiconductors Program and the Vanier Scholarship CGS program.
文摘Modern miniaturization and the digitalization of characterization instruments greatly facilitate the diffusion of technological advances in new fields and generate innovative applications.The concept of a portable,inexpensive and semi-automated biosensing platform,or lab-on-a-chip,is a vision shared by many researchers and venture industries.Under this scope,we present a semiconductor monolithic integration approach to conduct surface plasmon resonance studies.This technology is already commonly used for biochemical characterization in pharmaceutical industries,but we have reduced the technological platform to a few nanometers in scale on a semiconductor chip.We evaluate the signal quality of this nanophotonic device using hyperspectral-imaging technology,and we compare its performance with that of a standard prism-based commercial system.Two standard biochemical agents are employed for this characterization study:bovine serum albumin and inactivated influenza A virus.Time resolutions of data acquisition varying between 360 and 2.2 s are presented,yielding 2.731025–1.531026 RIU resolutions,respectively.
文摘A number of eukaryotic expression vectors have been developed for use as DNA vaccines. They showed varying abilities to initiate immune responses;however, there is little data to indicate which of these vectors will be the most useful and practical for DNA vaccines in different species. This report examines the use of five expression vectors with different promoters and Kozak sequence to express the same hemagglutinin (HA) protein of an H6N2 avian influenza virus for DNA vaccination in chickens. Although intramuscular vaccination with seven DNA constructs elicited no or limited measurable H6 HA antibody responses in Hy-Line chickens, variable reduction in virus shedding for either oropharyngeal or cloacal swabs post-virus challenge were observed. This indicated that all DNA constructs generated some levels of protective immunity against homologous virus challenge. Interestingly, lower dose (50 or 100 μg) of plasmid DNAs consistently induced better immune response than higher dose (300 or 500 μg). In the transfection experiments there appeared to be a hierarchy in the in vitro expression efficiency in the order of pCAG-optiHAk/ pCAG-HAk > pCI-HAk > VR-HA > pCI-HA > pCI-neo-HA > pVAX-HA. Since the level of in vitro expression correlates with the level of immune response in vivo, in vitro expression levels of the DNA constructs can be used as an indicator for pre-selection of plasmid vaccines prior to in vivo assessment. Moreover, our results suggested that the Kozak sequence could be used as an effective tool for DNA vaccine design.
基金partially supported by the National Institutes of Health(grant no.P20GM103646)the United States Department of Agriculture Animal and Plant Health Inspection Service(agreement 14-7428-1041-CA)
文摘Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).
