Influenza A Virus (IAV) binds sialic acid glycan receptor

GO:0030665

GO clathrin-coated vesicle membrane

PR:000049750

PR influenzavirus hemagglutinin

GO:0012506

GO vesicle membrane

PR:000044794

PR influenzavirus NS gene translation product

GO:0031381

GO viral RNA-directed RNA polymerase complex

GO:0019017

GO segmented viral genome

GO:0005102

GO receptor binding

GO:0036010

GO protein localization to endosome

GO:0006898

GO receptor-mediated endocytosis

GO:0061025

GO membrane fusion

GO:0034154

GO toll-like receptor 7 signaling pathway

GO:0034138

GO toll-like receptor 3 signaling pathway

GO:0046775

GO suppression by virus of host cytokine production

GO:0009616

GO virus induced gene silencing

GO:0075528

GO modulation by virus of host immune response

GO:0006606

GO protein import into nucleus

GO:0001172

GO transcription, RNA-templated

GO:0046784

GO viral mRNA export from host cell nucleus

GO:0019075

GO virus maturation

GO:0046761

GO viral budding from plasma membrane

GO:0019089

GO transmission of virus

WIKI occurrence

WIKI decreased

Influenza Virus

2021-12-05T12:32:16

WCS_9606

common toxicological species human

WCS_9031

common ecological species chicken

WikiUser_24

Wikiuser:Migration Pig

10090

NCBI mouse

9685

NCBI cat

WCS_9615

common toxicological species dog

9669

NCBI ferret

10036

NCBI Syrian hamster

10141

NCBI guinea pig

9544

NCBI rhesus macaque

WikiUser_25

Wikiuser: Cyauk human and other cells in culture

Influenza A Virus (IAV) binds sialic acid glycan receptor

IAV binds receptor

Molecular

Sialic acid was one of the first viral receptors identified5. Humans have 6 sialyl transferases that catalyze the addition of Sia with an a2,3 linkage to terminal galactose residues and 2 that catalyze the addition of an a2,6 linkage to terminal galactose residues6.   The HA receptor of the IAV attaches to the surface of the host cell via glycoconjugates that contain terminal sialic acid residues. The virus then “scans” the surface of the cell for the correct receptor, using its NA to remove nonproductive HA associations. The exact receptor is currently unknown however human influenza viruses preferentially bind sialic acid linked to galactose by a2,6 linkage, while avian influenza viruses prefer a2,3 linkages1. However, most viruses are not this dichotomous and the ability to bind sialic acid is more of a spectrum2. Additionally, the human respiratory tract contains both types of linkages as a gradient, with more a2,6 linked sialic acids present in the upper airway transitioning to more a2,3 linked sialic acids in the lower airway3. Some avian viruses can only replicate effectively in cells that express a2,3 linked sialic acids, which in humans is limited to the lower respiratory tract, which may serve as barrier to interspecies transmission and require that successful zoonosis is contingent upon the ability of the virus to bind a2,6 linked sialic acids, making this a marker of pandemic potential3. However, this is complicated by new evidence that non-binding sialic acids can contribute to enhanced binding and infection through hetero-multivalent interactions4. Individual hemagglutinin (HA) interactions with sialic acid glycan receptors are low affinity (KD ~0,5 to 20mM) leading to a low initial binding rate but high avidity is achieved through multivalent interactions with a receptor coated surface4.   Recent findings suggest phosphor-glycans are a potential alternative IAV receptor7. Additionally, two subtypes of IAV found exclusively in South and Central American bats (H17N10 and H18N11) use MHC class II for entry7,8.

Several studies have determined a dissociation constant (KD) for IAV and sialic acid glycan receptors as follows: <table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal; width:671px"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; height:23px; vertical-align:top; width:273px"> Reference </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:23px; vertical-align:top; width:131px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:23px; vertical-align:top; width:164px"> Binding partner </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:23px; vertical-align:top; width:103px"> Measured Kd </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:273px"> Sauter, N. K. et al. Hemagglutinins from two influenza virus variants bind to sialic acid derivatives with millimolar dissociation constants: a 500-MHz proton nuclear magnetic resonance study. Biochemistry 28, 8388–8396 (1989). </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:131px"> 500-MHz proton nuclear magnetic resonance (NMR) </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:164px"> X-31BHA virus (H3N2) with a(2,3)-Sialyl- lactose </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:103px"> 3.2 mM </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:273px"> Xiong, X., Coombs, P., Martin, S. et al. Receptor binding by a ferret-transmissible H5 avian influenza virus. Nature 497, 392–396 (2013). https://doi.org/10.1038/nature12144 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:131px"> microscale thermophoresis (MST) using recombinant HA trimers and surface biolayer interferometry (BLI) with purified viruses </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:164px"> A/Vietnam/1194/2004 (H5N1) with human and avian receptor </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:103px"> Human: 17mM   Avian: 1.1mM </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:273px"> Fei, Y. et al. Characterization of Receptor Binding Profiles of Influenza A Viruses Using An Ellipsometry-Based Label-Free Glycan Microarray Assay Platform. Biomolecules 5, 1480–1498 (2015). </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:131px"> Glycan microarray with a scanning ellipsometry sensor </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:164px"> A/Memphis/1971 (A/Mem71, H3N1), A/Udorn/307/1972 (A/Udorn72, H3N2), and A/Philippines/2/82/X-79 (A/Philips, H3N2) with 24 synthetic glycans (oligosaccharides) including include four β1-4-linked galactosides, three β1-3-linked galactosides, one β-linked galactoside, one α-linked N-acetylgalactosaminide, eight α2-3-linked sialosides, and seven α2-6-linked sialosides </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:103px"> 100pM </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:273px"> Vachieri, S. G. et al. Receptor binding by H10 influenza viruses. Nature 511, 475–477 (2014). </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:131px"> Biolayer interferometry </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:164px"> H10 virus to human and avian receptor </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:80px; vertical-align:top; width:103px"> Avian: 1.81 ± 0.39 mM   Human: 1.39 ± 0.32 mM, </td> </tr> </tbody> </table>   Other studies have characterized this interaction to identify species specificity:   <table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:253px"> Reference </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:190px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:181px"> Finding </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Rogers, G., Paulson, J., Daniels, R. et al. Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity. Nature 304, 76–78 (1983). https://doi.org/10.1038/304076a0 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, HAI </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Specific mutations at site 226 in HA impact sialic acid linkage binding preference </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Rogers GN, Pritchett TJ, Lane JL, Paulson JC. Differential sensitivity of human, avian, and equine influenza A viruses to a glycoprotein inhibitor of infection: selection of receptor specific variants. Virology. 1983 Dec;131(2):394-408. doi: 10.1016/0042-6822(83)90507-x. PMID: 6197808. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, HAI </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Human, avian, and equine H3 Influenza A viruses have different abilities to bind sialic acid (human prefer 2,6, animals prefer 2,3). </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Childs, R., Palma, A., Wharton, S. et al. Receptor-binding specificity of pandemic influenza A (H1N1) 2009 virus determined by carbohydrate microarray. Nat Biotechnol 27, 797–799 (2009). https://doi.org/10.1038/nbt0909-797 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Carbohydrate microarray </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Pandemic viruses were able to bind both 2,6 and 2,3 linked sialyl glycans while seasonal viruses only bound 2,6 </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Matrosovich M, Tuzikov A, Bovin N, Gambaryan A, Klimov A, Castrucci MR, Donatelli I, Kawaoka Y. Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals. J Virol. 2000 Sep;74(18):8502-12. doi: 10.1128/jvi.74.18.8502-8512.2000. PMID: 10954551; PMCID: PMC116362. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Solid- phase receptor binding assay </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Alteration of receptor binding efficiency may be a prerequisite for zoonosis </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Crusat M, Liu J, Palma AS, Childs RA, Liu Y, Wharton SA, Lin YP, Coombs PJ, Martin SR, Matrosovich M, Chen Z, Stevens DJ, Hien VM, Thanh TT, Nhu le NT, Nguyet LA, Ha do Q, van Doorn HR, Hien TT, Conradt HS, Kiso M, Gamblin SJ, Chai W, Skehel JJ, Hay AJ, Farrar J, de Jong MD, Feizi T. Changes in the hemagglutinin of H5N1 viruses during human infection--influence on receptor binding. Virology. 2013 Dec;447(1-2):326-37. doi: 10.1016/j.virol.2013.08.010. Epub 2013 Sep 17. PMID: 24050651; PMCID: PMC3820038. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, receptor binding assay using sialylglycopolymers, biolayer interferometry analysis, carbohydrate microarray analysis, crystallography </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> H5N1 infection of human leads to decreased ability to bind 2,3 linked sialic acid </td> </tr> </tbody> </table>

UBERON:0004802

UBERON respiratory tract epithelium

CL:0000066

CL epithelial cell

High Mixed

High

Adult, reproductively mature

High

During development and at adulthood

High High High High Moderate Moderate High High High High

References: <ol> <li>Paulson, J. and Rogers, G. Receptor determinants of human and animal influenza virus isolates: Differences in recptor specificity of the H3 hemagglutinin based on species of origin. Virology 127:2, 361-373 (1983). <a href="https://doi.org/10.1016/0042-6822(83)90150-2" style="color:#954f72; text-decoration:underline" target="_blank" title="Persistent link using digital object identifier">https://doi.org/10.1016/0042-6822(83)90150-2</a></li> <li>Get this from thesis</li> <li>Shinya, K., Ebina, M., Yamada, S. et al. Influenza virus receptors in the human airway. Nature 440, 435–436 (2006). <a href="https://doi.org/10.1038/440435a" style="color:#954f72; text-decoration:underline">https://doi.org/10.1038/440435a</a></li> <li>Liu, M., Huang, L.Z.X., Smits, A.A. et al. Human-type sialic acid receptors contribute to avian influenza A virus binding and entry by hetero-multivalent interactions. Nat Commun13, 4054 (2022). <a href="https://doi.org/10.1038/s41467-022-31840-0" style="color:#954f72; text-decoration:underline">https://doi.org/10.1038/s41467-022-31840-0</a></li> <li>Matrosovich M, Herrler G, Klenk HD. Sialic Acid Receptors of Viruses. Top Curr Chem. 2015;367:1-28. doi: 10.1007/128_2013_466. PMID: 23873408; PMCID: PMC7120183.</li> <li>Human Protein Atlas proteinatlas.org</li> <li>Sempere Borau, M and Stertz, S Entry of Influenza A virus into host cells- recent progress and remaining challenges. Current Opinion in Virology. 2021 doi: <a href="https://doi.org/10.1016/j.coviro.2021.03.001" rel="noreferrer noopener" target="_blank" title="Persistent link using digital object identifier">https://doi.org/10.1016/j.coviro.2021.03.001</a></li> <li>Karakus, U., Thamamongood, T., Ciminski, K. et al. MHC class II proteins mediate cross-species entry of bat influenza viruses. Nature 567, 109–112 (2019). https://doi.org/10.1038/s41586-019-0955-3</li> </ol> AOPWiki 2023-07-31T13:37:09

2023-08-02T10:27:29

Influenza A virus (IAV) cell entry

IAV cell entry

Cellular

IAV has two major surface proteins: hemagglutinin (HA) and neuraminidase (NA). HA binds to sialic acid glycans on the host cell surface to facilitate viral entry (1,2). Following this, the virion enters the cell through receptor—mediated endocytosis (usually involving clathrin) or micropinocytosis (1,3,4). The virus is then trafficked to the endosome, where the change in pH activates the M2 ion channel protein of the virus, leading to a conformational change in the HA exposing the fusion peptide and causing subsequent fusion of the viral envelope with the membrane of the vesicle (1). Following fusion, the vRNPs are released into the cytoplasm in a process known as “uncoating” and trafficked to the nucleus (1). This entire process takes about 10 minutes (5)

<table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Reference </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Finding </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Matlin, K.S., Reggio, H., Helenius, A. & Simons, K. Infectious entry pathway of influenza-virus in a canine kidney-cell line. J. Cell Biol. 91, 601–613 (1981) </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Electron microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Virus was seen bound to microvilli, in coated pits, coated vesicles, and large smooth-surfaced vacuoles, low pH was required for fusion, suggesting entry by endocytosis </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Rust, M., Lakadamyali, M., Zhang, F. et al. Assembly of endocytic machinery around individual influenza viruses during viral entry. Nat Struct Mol Biol 11, 567–573 (2004). https://doi.org/10.1038/nsmb769 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Real- time fluorescent microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Clathrin-mediated and clathrin- and caveolin-independent endocytic pathways used in parallel with similar efficiency </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> De Vries, E. et. al., Dissection of the Influenza A Virus Endocytic Routes Reveals Macropinocytosis as an Alternative Entry Pathway. Plos Pathogens (2011). <a href="https://doi.org/10.1371/journal.ppat.1001329" style="color:#954f72; text-decoration:underline">https://doi.org/10.1371/journal.ppat.1001329</a>   </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Luciferase reporter assay </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Macropinocytosis is an alternative entry pathway </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Chen, C. and Zhuang, X. Epsin 1 is a cargo- specific adaptor for the clathrin-mediated endocytosis of the influenza virus </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Colocalization of immunofluorescence </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> influenza entry via clathrin- mediated pathway   </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Sieczkarski, S. and Whittaker, G. Influenza Virus Can Enter and Infect Cells in the Absence of Clathrin-Mediated Endocytosis </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Flow cytommetry </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> IAV cell entry via non-clathrin dependent route </td> </tr> </tbody> </table>

UBERON:0000065

UBERON respiratory tract

CL:0002368

CL respiratory epithelial cell

High Unspecific

Moderate

All life stages

High High

<ol> <li>Dou, D., et. al. Influenza A Virus Cell Entry, Replication, Virion Assembly, and Movement. Front. Immunol. (2018) <a href="https://doi.org/10.3389/fimmu.2018.01581" style="color:#954f72; text-decoration:underline">https://doi.org/10.3389/fimmu.2018.01581</a></li> <li>Sempere Borau, M. and Stertz, S. Entry of influenza A virus into host cells- recent progress and remaining challenges. Current Opinion in Virology (2021) <a href="https://doi.org/10.1016/j.coviro.2021.03.001" style="color:#954f72; text-decoration:underline">https://doi.org/10.1016/j.coviro.2021.03.001</a></li> <li>Matlin, K.S., Reggio, H., Helenius, A. & Simons, K. Infectious entry pathway of influenza-virus in a canine kidney-cell line. J. Cell Biol. 91, 601–613 (1981) <a href="https://doi.org/10.1083/jcb.91.3.601" style="color:#954f72; text-decoration:underline">https://doi.org/10.1083/jcb.91.3.601</a></li> <li>De Vries, E. et. al., Dissection of the Influenza A Virus Endocytic Routes Reveals Macropinocytosis as an Alternative Entry Pathway. Plos Pathogens (2011). <a href="https://doi.org/10.1371/journal.ppat.1001329" style="color:#954f72; text-decoration:underline">https://doi.org/10.1371/journal.ppat.1001329</a></li> <li>Dou D, Hernandez-Neuta I, Wang H, Ostbye H, Qian X, Thiele S, et al. Analysis of IAV replication and co-infection dynamics by a versatile RNA viral genome labeling method. Cell Rep (2017) 20:251–63. doi:10.1016/j.celrep.2017.06.021</li> </ol> AOPWiki 2023-08-11T12:25:13

2023-10-03T10:16:57

Immune mechanisms antagonized by viral proteins

immune mechanisms antagonized

Molecular

IAV infection is detected by multiple host sensors. Within the infected cell, viral RNA in the cytosol (potentially as part of a stress granule) is recognized by RIG-I which signals through MAVS to induce pro-inflammatory cytokines and type 1 IFN (1). Additionally, while toll-like receptor (TLR) expression is usually localized to immune cells, nasal epithelial cells also express a majority of TLR subtypes (2). This allows for recognition of IAV through TLR3 or TLR7(1). Sensing of dsRNA through TLR3 and TRIF intermediate leads to a signaling cascade via NFkB and IRF3 to induce expression of Type I IFNs and ISGs as well as IL-1B and other pro-inflammatory cytokines (1). This is despite the fact that IAV does not produce dsRNA, however due to interactions with UAP56, TLR3 is able to recognize different RNA structures (5). Sensing ssRNA through TLR7 and Myd88 intermediate leads to a signaling cascade involving NFkB and IRF7 to also induce expression of Type I IFNs and ISGs as well as pro-inflammatory cytokines (1). The IAV NS1 protein binds to CPSF30 blocking the cleavage of pre-mRNAs and recruitment of the poly(A) polymerase and is considered the key antagonizing factor (3,4). This causes accumulation of unprocessed cellular pre-mRNA accumulation in the nucleus leading to the inhibition of general gene expression as well as induction of IFN, ISGs, and other pro-inflammatory gene (3). Additionally, NS1 can bind cellular dsDNA as well as components of mRNA export machinery, again preventing the expression of antiviral genes (3).Additionally, the IAV PA-X protein also blocks cellular antiviral responses by selectively degrading host RNA Pol II transcribed RNAs in the nucleus, sparing Pol I and Pol III products (3). This again inhibits expression of antiviral and proinflammatory genes.

Current research (mostly in mouse models and from clinical data anaylsis) suggests that sex differences in response to infection emerge during the adaptive immune response. The innate immune response steps outlined in this KE are expected to be universal, but the magnitude and some mechanisms may differ between sexes and more work is needed to understand (6-8)

UBERON:0000065

UBERON respiratory tract

CL:0002368

CL respiratory epithelial cell

Moderate Unspecific

High

Adult, reproductively mature

High High

<ol> <li>Iwasaki, A., Pillai, P. Innate immunity to influenza virus infection. Nat Rev Immunol 14, 315–328 (2014). <a href="https://doi.org/10.1038/nri3665" style="color:#954f72; text-decoration:underline">https://doi.org/10.1038/nri3665</a></li> <li>McClure, R. and Massari, P. TLR-dependent human mucosal epithelial responses to microbial pathogens. Front. Immunol. (2014). doi: <a href="https://doi.org/10.3389%2Ffimmu.2014.00386" style="color:#954f72; text-decoration:underline" target="_blank">10.3389/fimmu.2014.00386</a></li> <li>Nogales, A., et. al., Modulation of Innate Immune Responses by the Influenza A NS1 and PA-X Proteins. MDPI viruses. (2018). doi: <a href="https://doi.org/10.3390%2Fv10120708" style="color:#954f72; text-decoration:underline" target="_blank">10.3390/v10120708</a></li> <li>Zhang, Y., Xu, Z., and Cao, Y., Host-Virus Interaction: How Host Cells defend against Influenza A Virus Infection. MDPI viruses. (2020). doi: <a href="https://doi.org/10.3390%2Fv12040376" style="color:#954f72; text-decoration:underline" target="_blank">10.3390/v12040376</a></li> <li>Schulz, O., Diebold, S., Chen, M. et al. Toll-like receptor 3 promotes cross-priming to virus-infected cells. Nature 433, 887–892 (2005). https://doi.org/10.1038/nature03326</li> <li>Peretz J, Pekosz A, Lane AP, Klein SL. Estrogenic compounds reduce influenza A virus replication in primary human nasal epithelial cells derived from female, but not male, donors. Am J Physiol Lung Cell Mol Physiol. 2016 Mar 1;310(5):L415-25. doi: 10.1152/ajplung.00398.2015. Epub 2015 Dec 18. PMID: 26684252; PMCID: PMC4773846.</li> <li>Klein SL, Flanagan KL. Sex differences in immune responses. Nat Rev Immunol. 2016 Oct;16(10):626-38. doi: 10.1038/nri.2016.90. Epub 2016 Aug 22. PMID: 27546235.</li> <li>Jacobsen H, Klein SL. Sex Differences in Immunity to Viral Infections. Front Immunol. 2021 Aug 31;12:720952. doi: 10.3389/fimmu.2021.720952. PMID: 34531867; PMCID: PMC8438138.</li> </ol> AOPWiki 2023-08-23T09:36:12

2023-12-13T10:05:20

Influenza A Virus production increased

IAV production increased

Molecular

The major stages of Influenza A virus (IAV) replication, which could each be multiple KE's on their own but are summarized here, include trafficking to the host cell nucleus, replication of vRNAs, viral mRNA transcription, assembly and trafficking of vRNPs, ER targeting and maturation, and HA proteolytic activation at the Golgi or Plasma membrane (1).  The current model of IAV replication is vRNP entry into the nucleus via the importin-alpha-importin-beta nuclear import pathway within one hour (1-9). However, this step has been shown to contribute to host restriction so it is worth noting that a majority of this work has been performed in immortalized cell lines from various species (1). Replication of vRNAs occurs in two steps- transcription of the reverse complement (cRNA) followed by transcription of new genome copies using the cRNAs as templates- in the nucleus facilitated by the heterotrimeric viral RNA-dependent RNA polymerase (1). The transcription process is unprimed and reliant on 13 conserved nucleotides at the 5’ end and 12 nucleotides at the 3’ end of each segment that are partially complementary and form a double-stranded “promoter” by base-pairing (10). This “promoter” is recognized by the viral RNA polymerase and the template is transcribed. Viral mRNA transcription, in contrast, is primed by a process known as “cap- snatching” from host mRNAs (11,12) and polyadenylated through stuttering (13,14). Two segments- M and NS- are spliced (15). NS transcripts have a balanced ratio of spliced to un spliced throughout infection but the ratio of spliced M (M2) increases through infection, although the efficiency differs between strains and also functions in host restriction (16-19). Following nuclear export, translation of the viral mRNAs is divided between cytosolic ribosomes (internal proteins PB1, PB2, PA, NP, NS1, NS2, and M1) and ER- associated ribosomes (membrane proteins- HA, NA, and M2) (1). The NP and polymerase subunits (PB1, PB2, PA) traffic back to the nucleus to assist in viral mRNA transcription and vRNA replication (1) while NS1 is also trafficked to the nucleus to inhibit interferon signaling (see KE 2180). vRNPs are trafficked to the plasma membrane to meet the other proteins for viral assembly by Rab11, which associates with Pb2 to ensure that all new virions incorporated vRNPs carrying a polymerase (20). During translocation within the ER, the N-terminus of HA and M2 is directly translocated into the ER lumen, while the NA C-terminus is positioned in the ER lumen (1). Both HA and NA receive multiple N-linked glycans which have been shown to affect activity and antigenicity but mainly function in folding efficiency (21-24). The HA protein is formed through trimerization of independently folded monomers while the NA tetramer results from association of co-translationally formed dimers (21,25,26,27). An abundance of HA and NA is produced during infection to promote oligomerization but consequences of this approach include triggering of the ER stress response, which must be blocked, and syncytia formation, which promotes cell to cell transmission (1,28,29,30) The HA surface protein is synthesized as an inactive form denoted HA0 (1, 31, 32, 33). HA0 is then proteolytically activated through cleavage into HA1 and HA2 active subunits by trypsin-like proteases (1,31,32,33,34). IAV is fairly promiscuous in its use of these proteases, and the cleavage occurs in a multi- or mono- basic site on the HA0 protein (34,35). Highly pathogenic avian IAVs tend to have a multibasic cleavage site that is targeted by furin (36). Human and low pathogenic avian viruses tend to contain a monobasic cleavage site and utilize any available protease such as TMRPSS2, TMPRSS4, and HAT for proteolytic activation (37,38).

<table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Conclusions/ uses </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Example Reference </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Immunofluorescence microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Visualization of infected cells, can target specific proteins or processes </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> <ol> <li>Martin K, Helenius A. Transport of incoming influenza virus nucleocapsids into the nucleus. J Virol (1991) 65:232–44.</li> <li>O’Neill RE, Jaskunas R, Blobel G, Palese P, Moroianu J. Nuclear import of influenza virus RNA can be mediated by viral nucleoprotein and transport factors required for protein import. J Biol Chem (1995) 270:22701–4. doi:10.1074/jbc.270.39.22701</li> </ol>   </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> In situ hybridization </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Track localization of RNA viral genomes in cells </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Dou D, Hernandez-Neuta I, Wang H, Ostbye H, Qian X, Thiele S, et al. Analysis of IAV replication and co-infection dynamics by a versatile RNA viral genome labeling method. Cell Rep (2017) 20:251–63. doi:10.1016/j.celrep.2017.06.021 </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> In vitro polyadenylation assay </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Identify sequences important for polyadenylation </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Poon LL, Pritlove DC, Fodor E, Brownlee GG. Direct evidence that the poly(A) tail of influenza A virus mRNA is synthesized by reiterative copying of a U track in the virion RNA template. J Virol (1999) 73:3473–6. </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Plasmid expression measured by Northern Blot </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Analyze differences in splicing efficiencies </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Backstrom Winquist E, Abdurahman S, Tranell A, Lindstrom S, Tingsborg S, Schwartz S. Inefficient splicing of segment 7 and 8 mRNAs is an inherent property of influenza virus A/Brevig Mission/1918/1 (H1N1) that causes elevated expression of NS1 protein. Virology (2012) 422:46–58. doi:10.1016/j.virol.2011.10.004   </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Single molecule FRET </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Molecule- molecule interactions </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Robb NC, Te Velthuis AJ, Wieneke R, Tampé R, Cordes T, Fodor E, Kapanidis AN. Single-molecule FRET reveals the pre-initiation and initiation conformations of influenza virus promoter RNA. Nucleic Acids Res. 2016 Dec 1;44(21):10304-10315. doi: 10.1093/nar/gkw884. Epub 2016 Sep 30. PMID: 27694620; PMCID: PMC5137447. </td> </tr> </tbody> </table>

UBERON:0000065

UBERON respiratory tract

CL:0002368

CL respiratory epithelial cell

<ol> <li>Dou, D., Revol, R., Ostbye, H., Wang, H., and Daniels, R., Influenza A Virus Cell Entry, Replication, Virion Assembly and Movement. Front. Immunol.  20 July 2018. <a href="https://doi.org/10.3389/fimmu.2018.01581" style="color:#954f72; text-decoration:underline">https://doi.org/10.3389/fimmu.2018.01581</a></li> <li>Martin K, Helenius A. Transport of incoming influenza virus nucleocapsids into the nucleus. J Virol (1991) 65:232–44.</li> <li>Kemler I, Whittaker G, Helenius A. Nuclear import of microinjected influenza virus ribonucleoproteins. Virology (1994) 202:1028–33. doi:10.1006/viro.1994.1432</li> <li>O’Neill RE, Jaskunas R, Blobel G, Palese P, Moroianu J. Nuclear import of influenza virus RNA can be mediated by viral nucleoprotein and transport factors required for protein import. J Biol Chem (1995) 270:22701–4. doi:10.1074/jbc.270.39.22701</li> <li>Wang P, Palese P, O’Neill RE. The NPI-1/NPI-3 (karyopherin alpha) binding site on the influenza a virus nucleoprotein NP is a nonconventional nuclear localization signal. J Virol (1997) 71:1850–6.</li> <li>Cros JF, Garcia-Sastre A, Palese P. An unconventional NLS is critical for the nuclear import of the influenza A virus nucleoprotein and ribonucleoprotein. Traffic (2005) 6:205–13. doi:10.1111/j.1600-0854.2005.00263.x</li> <li>Wu WW, Weaver LL, Pante N. Ultrastructural analysis of the nuclear localization sequences on influenza A ribonucleoprotein complexes. J Mol Biol (2007) 374:910–6. doi:10.1016/j.jmb.2007.10.022</li> <li>Chou YY, Heaton NS, Gao Q, Palese P, Singer RH, Lionnet T. Colocalization of different influenza viral RNA segments in the cytoplasm before viral budding as shown by single-molecule sensitivity FISH analysis. PLoS Pathog (2013) 9:e1003358. doi:10.1371/journal.ppat.1003358</li> <li>Dou D, Hernandez-Neuta I, Wang H, Ostbye H, Qian X, Thiele S, et al. Analysis of IAV replication and co-infection dynamics by a versatile RNA viral genome labeling method. Cell Rep (2017) 20:251–63. doi:10.1016/j.celrep.2017.06.021</li> <li>Robb NC, Te Velthuis AJ, Wieneke R, Tampe R, Cordes T, Fodor E, et al. Single-molecule FRET reveals the pre-initiation and initiation conformations of influenza virus promoter RNA. Nucleic Acids Res (2016) 44:10304–15. doi:10.1093/nar/gkw884</li> <li>Reich S, Guilligay D, Pflug A, Malet H, Berger I, Crepin T, et al. Structural insight into cap-snatching and RNA synthesis by influenza polymerase. Nature (2014) 516:361–6. doi:10.1038/nature14009</li> <li>Plotch SJ, Bouloy M, Ulmanen I, Krug RM. A unique cap(m7GpppXm)-dependent influenza virion endonuclease cleaves capped RNAs to generate the primers that initiate viral RNA transcription. Cell (1981) 23:847–58. doi:10.1016/0092-8674(81)90449-9</li> <li>Robertson JS, Schubert M, Lazzarini RA. Polyadenylation sites for influenza virus mRNA. J Virol (1981) 38:157–63.</li> <li>Poon LL, Pritlove DC, Fodor E, Brownlee GG. Direct evidence that the poly(A) tail of influenza A virus mRNA is synthesized by reiterative copying of a U track in the virion RNA template. J Virol (1999) 73:3473–6.</li> <li>Dubois J, Terrier O, Rosa-Calatrava M. Influenza viruses and mRNA splicing: doing more with less. MBio (2014) 5:e70–14. doi:10.1128/mBio.00070-14</li> <li>Inglis SC, Brown CM. Differences in the control of virus mRNA splicing during permissive or abortive infection with influenza A (fowl plague) virus. J Gen Virol (1984) 65(Pt 1):153–64. doi:10.1099/0022-1317-65-1-153</li> <li>Valcarcel J, Portela A, Ortin J. Regulated M1 mRNA splicing in influenza virus-infected cells. J Gen Virol (1991) 72(Pt 6):1301–8. doi:10.1099/0022-1317-72-6-1301</li> <li>Robb NC, Fodor E. The accumulation of influenza A virus segment 7 spliced mRNAs is regulated by the NS1 protein. J Gen Virol (2012) 93:113–8. doi:10.1099/vir.0.035485-0</li> <li>Backstrom Winquist E, Abdurahman S, Tranell A, Lindstrom S, Tingsborg S, Schwartz S. Inefficient splicing of segment 7 and 8 mRNAs is an inherent property of influenza virus A/Brevig Mission/1918/1 (H1N1) that causes elevated expression of NS1 protein. Virology (2012) 422:46–58. doi:10.1016/j.virol.2011.10.004</li> <li>Amorim MJ, Bruce EA, Read EK, Foeglein A, Mahen R, Stuart AD, et al. A Rab11- and microtubule-dependent mechanism for cytoplasmic transport of influenza A virus viral RNA. J Virol (2011) 85:4143–56. doi:10.1128/JVI.02606-10</li> <li>Wang N, Glidden EJ, Murphy SR, Pearse BR, Hebert DN. The cotranslational maturation program for the type II membrane glycoprotein influenza neuraminidase. J Biol Chem (2008) 283:33826–37. doi:10.1074/jbc.M806897200</li> <li> Daniels R, Svedine S, Hebert DN. N-linked carbohydrates act as lumenal maturation and quality control protein tags. Cell Biochem Biophys (2004) 41:113–38. doi:10.1385/CBB:41:1:113</li> <li>Hebert DN, Zhang JX, Chen W, Foellmer B, Helenius A. The number and location of glycans on influenza hemagglutinin determine folding and association with calnexin and calreticulin. J Cell Biol (1997) 139:613–23. doi:10.1083/jcb.139.3.613</li> <li>Powell H,  Pekosz A. Neuraminidase antigenic drift of H3N2 clade 3c.2a viruses alters virus replication, enzymatic activity, and inhibitory antibody binding. Plos Pathogens (2020). <a href="https://doi.org/10.1371/journal.ppat.1008411" style="color:#954f72; text-decoration:underline">https://doi.org/10.1371/journal.ppat.1008411</a></li> <li>Saito T, Taylor G, Webster RG. Steps in maturation of influenza A virus neuraminidase. J Virol (1995) 69:5011–7.</li> <li>da Silva DV, Nordholm J, Madjo U, Pfeiffer A, Daniels R. Assembly of subtype 1 influenza neuraminidase is driven by both the transmembrane and head domains. J Biol Chem (2013) 288:644–53. doi:10.1074/jbc.M112.424150</li> <li>da Silva DV, Nordholm J, Dou D, Wang H, Rossman JS, Daniels R. The influenza virus neuraminidase protein transmembrane and head domains have coevolved. J Virol (2015) 89:1094–104. doi:10.1128/JVI.02005-14</li> <li>Hassan IH, Zhang MS, Powers LS, Shao JQ, Baltrusaitis J, Rutkowski DT, et al. Influenza A viral replication is blocked by inhibition of the inositol-requiring enzyme 1 (IRE1) stress pathway. J Biol Chem (2012) 287:4679–89. doi:10.1074/jbc.M111.284695</li> <li>Roberson EC, Tully JE, Guala AS, Reiss JN, Godburn KE, Pociask DA, et al. Influenza induces endoplasmic reticulum stress, caspase-12-dependent apoptosis, and c-Jun N-terminal kinase-mediated transforming growth factor-beta release in lung epithelial cells. Am J Respir Cell Mol Biol (2012) 46:573–81. doi:10.1165/rcmb.2010-0460OC</li> <li>Cifuentes-Muñoz N, Dutch RE, Cattaneo R. Direct cell-to-cell transmission of respiratory viruses: The fast lanes. PLoS Pathog. 2018 Jun 28;14(6):e1007015. doi: 10.1371/journal.ppat.1007015. PMID: 29953542; PMCID: PMC6023113.</li> <li>Klenk HD, Rott R, Orlich M, Blodorn J.Activation of influenza A viruses by trypsin treatment. Virology (1975) 68:426–39. 10.1016/0042-6822(75)90284-6</li> <li>Huang RT, Rott R, Klenk HD. Influenza viruses cause hemolysis and fusion of cells. Virology (1981) 110:243–7. 10.1016/0042-6822(81)90030-1</li> <li>Maeda T, Kawasaki K, Ohnishi S. Interaction of influenza virus hemagglutinin with target membrane lipids is a key step in virus-induced hemolysis and fusion at pH 5.2. Proc Natl Acad Sci U S A (1981) 78:4133–7. 10.1073/pnas.78.7.4133</li> <li>Kido,  H., Okumura,  Y., Yamada,  H., Quang Le,  T. & Yano, M. Proteases essential for human influenza virus entry into cells and their inhibitors as potential therapeutic agents. Curr Pharm Des 13, 405–414 (2007))</li> <li>Bottcher-Friebertshauser E, Garten W, Matrosovich M, Klenk HD. The hemagglutinin: a determinant of pathogenicity. Curr Top Microbiol Immunol (2014) 385:3–34. 10.1007/82_2014_384 </li> <li>Stieneke-Grober A, Vey M, Angliker H, Shaw E, Thomas G, Roberts C, et al. Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin-like endoprotease. EMBO J (1992) 11:2407–14.</li> <li>Bottcher E, Matrosovich T, Beyerle M, Klenk HD, Garten W, Matrosovich M. Proteolytic activation of influenza viruses by serine proteases TMPRSS2 and HAT from human airway epithelium. J Virol (2006) 80:9896–8. 10.1128/JVI.01118-06 </li> <li>Chaipan C, Kobasa D, Bertram S, Glowacka I, Steffen I, Tsegaye TS, et al. Proteolytic activation of the 1918 influenza virus hemagglutinin. J Virol (2009) 83:3200–11. 10.1128/JVI.02205-08</li> <li>Lingwood D, Simons K. Lipid rafts as a membrane-organizing principle. Science (2010) 327:46–50. 10.1126/science.1174621</li> <li>Chen BJ, Leser GP, Morita E, Lamb RA. Influenza virus hemagglutinin and neuraminidase, but not the matrix protein, are required for assembly and budding of plasmid-derived virus-like particles. J Virol (2007) 81:7111–23. 10.1128/JVI.00361-07</li> <li>Lai JC, Chan WW, Kien F, Nicholls JM, Peiris JS, Garcia JM. Formation of virus-like particles from human cell lines exclusively expressing influenza neuraminidase. J Gen Virol (2010) 91:2322–30. 10.1099/vir.0.019935-0</li> <li>Yondola MA, Fernandes F, Belicha-Villanueva A, Uccelini M, Gao Q, Carter C, et al. Budding capability of the influenza virus neuraminidase can be modulated by tetherin. J Virol (2011) 85:2480–91. 10.1128/JVI.02188-10</li> <li>Chlanda P, Schraidt O, Kummer S, Riches J, Oberwinkler H, Prinz S, et al. Structural analysis of the roles of influenza A virus membrane-associated proteins in assembly and morphology. J Virol (2015) 89:8957–66. 10.1128/JVI.00592-15</li> <li>Elleman CJ, Barclay WS. The M1 matrix protein controls the filamentous phenotype of influenza A virus. Virology (2004) 321:144–53. 10.1016/j.virol.2003.12.009</li> <li>Rossman JS, Lamb RA. Viral membrane scission. Annu Rev Cell Dev Biol (2013) 29:551–69. 10.1146/annurev-cellbio-101011-155838</li> <li>Webster RG, Laver WG. Preparation and properties of antibody directed specifically against the neuraminidase of influenza virus. J Immunol(1967) 99:49–55.</li> <li>Palese P, Compans RW. Inhibition of influenza virus replication in tissue culture by 2-deoxy-2,3-dehydro-N-trifluoroacetylneuraminic acid (FANA): mechanism of action. J Gen Virol (1976) 33:159–63. 10.1099/0022-1317-33-1-159</li> </ol> AOPWiki 2023-10-02T14:16:23

2024-01-29T11:00:20

Influenza A Virus (IAV) budding

IAV budding

Cellular

IAV buds from lipid “rafts” present on the apical membrane (2). Once all the viral components have localized to the site of budding, curvature of the membrane is induced using a combination of molecular crowding and bending proteins. NA and HA proteins are sufficient to induce budding, and the M1 protein contributes to shape and size uniformity of the resulting virions (1,3,4,5,6,7). The M2 protein has also been shown to contribute to membrane bending and scission (8). After budding, release is mediated by the sialidase property of NA. The NA protein catalyzes the hydrolysis of all sialic acid attachments made by the HA protein allowing for the virus to be released from the cell membrane (1,9,10).

<table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Conclusions/ uses </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:208px"> Example Reference </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Virus- like particles </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Measure specific contributions of proteins to the viral life cycle </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> <ol> <li>Chen BJ, Leser GP, Morita E, Lamb RA. Influenza virus hemagglutinin and neuraminidase, but not the matrix protein, are required for assembly and budding of plasmid-derived virus-like particles. J Virol (2007) 81:7111–23. 10.1128/JVI.00361-07</li> <li>Lai JC, Chan WW, Kien F, Nicholls JM, Peiris JS, Garcia JM. Formation of virus-like particles from human cell lines exclusively expressing influenza neuraminidase. J Gen Virol (2010) 91:2322–30. 10.1099/vir.0.019935-0</li> </ol>   </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Electron microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Visualize budding </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:208px"> Chlanda P, Schraidt O, Kummer S, Riches J, Oberwinkler H, Prinz S, et al. Structural analysis of the roles of influenza A virus membrane-associated proteins in assembly and morphology. J Virol (2015) 89:8957–66. 10.1128/JVI.00592-15   </td> </tr> </tbody> </table>

UBERON:0000065

UBERON respiratory tract

CL:0000066

CL epithelial cell

<ol> <li>Dou, D., Revol, R., Ostbye, H., Wang, H., and Daniels, R., Influenza A Virus Cell Entry, Replication, Virion Assembly and Movement. Front. Immunol.  20 July 2018. <a href="https://doi.org/10.3389/fimmu.2018.01581">https://doi.org/10.3389/fimmu.2018.01581</a></li> <li>Lingwood D, Simons K. Lipid rafts as a membrane-organizing principle. Science (2010) 327:46–50. 10.1126/science.1174621</li> <li>Chen BJ, Leser GP, Morita E, Lamb RA. Influenza virus hemagglutinin and neuraminidase, but not the matrix protein, are required for assembly and budding of plasmid-derived virus-like particles. J Virol (2007) 81:7111–23. 10.1128/JVI.00361-07</li> <li>Lai JC, Chan WW, Kien F, Nicholls JM, Peiris JS, Garcia JM. Formation of virus-like particles from human cell lines exclusively expressing influenza neuraminidase. J Gen Virol (2010) 91:2322–30. 10.1099/vir.0.019935-0</li> <li>Yondola MA, Fernandes F, Belicha-Villanueva A, Uccelini M, Gao Q, Carter C, et al. Budding capability of the influenza virus neuraminidase can be modulated by tetherin. J Virol (2011) 85:2480–91. 10.1128/JVI.02188-10</li> <li>Chlanda P, Schraidt O, Kummer S, Riches J, Oberwinkler H, Prinz S, et al. Structural analysis of the roles of influenza A virus membrane-associated proteins in assembly and morphology. J Virol (2015) 89:8957–66. 10.1128/JVI.00592-15</li> <li>Elleman CJ, Barclay WS. The M1 matrix protein controls the filamentous phenotype of influenza A virus. Virology (2004) 321:144–53. 10.1016/j.virol.2003.12.009</li> <li>Rossman JS, Lamb RA. Viral membrane scission. Annu Rev Cell Dev Biol (2013) 29:551–69. 10.1146/annurev-cellbio-101011-155838</li> <li>Webster RG, Laver WG. Preparation and properties of antibody directed specifically against the neuraminidase of influenza virus. J Immunol(1967) 99:49–55.</li> <li>Palese P, Compans RW. Inhibition of influenza virus replication in tissue culture by 2-deoxy-2,3-dehydro-N-trifluoroacetylneuraminic acid (FANA): mechanism of action. J Gen Virol (1976) 33:159–63. 10.1099/0022-1317-33-1-159</li> </ol> AOPWiki 2023-10-02T14:20:32

2024-01-29T11:07:47

Influenza A Virus (IAV) shedding and transmission

infection proliferation

Individual

AOPWiki 2023-10-02T14:24:35

2023-10-02T14:24:35

<upstream-id>0971b052-14bb-4c0d-a529-b3c9c221bcfe</upstream-id> <downstream-id>aba81357-008c-4064-8eac-a1dd7471917b</downstream-id> This KER deals with the evidence that binding of eh Influenza HA protein to sialic acid glycans leads to viral entry into the cell.

To develop this KER, the author has reviewed the literature available for Influenza A virus receptor preferences up to December 2023 to find evidence showing: 1. Receptor essentiality for viral infection (shown through host suceptibility/ tropism) 2. Mechanisms that support viral entry, including the use of proteases

Multiple studies have characterized virus-receptor interactions and entry pathways:   <table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium; color:#000000; font-style:normal; font-weight:400; text-align:start; text-decoration:none; white-space:normal"> <tbody> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:253px"> Reference </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:190px"> Technique </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:181px"> Finding </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Rogers, G., Paulson, J., Daniels, R. et al. Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity. Nature 304, 76–78 (1983). https://doi.org/10.1038/304076a0 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, HAI </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Specific mutations at site 226 in HA impact sialic acid linkage binding preference </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Rogers GN, Pritchett TJ, Lane JL, Paulson JC. Differential sensitivity of human, avian, and equine influenza A viruses to a glycoprotein inhibitor of infection: selection of receptor specific variants. Virology. 1983 Dec;131(2):394-408. doi: 10.1016/0042-6822(83)90507-x. PMID: 6197808. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, HAI </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Human, avian, and equine H3 Influenza A viruses have different abilities to bind sialic acid (human prefer 2,6, animals prefer 2,3). </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Childs, R., Palma, A., Wharton, S. et al. Receptor-binding specificity of pandemic influenza A (H1N1) 2009 virus determined by carbohydrate microarray. Nat Biotechnol 27, 797–799 (2009). https://doi.org/10.1038/nbt0909-797 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Carbohydrate microarray </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Pandemic viruses were able to bind both 2,6 and 2,3 linked sialyl glycans while seasonal viruses only bound 2,6 </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Matrosovich M, Tuzikov A, Bovin N, Gambaryan A, Klimov A, Castrucci MR, Donatelli I, Kawaoka Y. Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals. J Virol. 2000 Sep;74(18):8502-12. doi: 10.1128/jvi.74.18.8502-8512.2000. PMID: 10954551; PMCID: PMC116362. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Solid- phase receptor binding assay </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Alteration of receptor binding efficiency may be a prerequisite for zoonosis </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Crusat M, Liu J, Palma AS, Childs RA, Liu Y, Wharton SA, Lin YP, Coombs PJ, Martin SR, Matrosovich M, Chen Z, Stevens DJ, Hien VM, Thanh TT, Nhu le NT, Nguyet LA, Ha do Q, van Doorn HR, Hien TT, Conradt HS, Kiso M, Gamblin SJ, Chai W, Skehel JJ, Hay AJ, Farrar J, de Jong MD, Feizi T. Changes in the hemagglutinin of H5N1 viruses during human infection--influence on receptor binding. Virology. 2013 Dec;447(1-2):326-37. doi: 10.1016/j.virol.2013.08.010. Epub 2013 Sep 17. PMID: 24050651; PMCID: PMC3820038. </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Hemagglutination assay, receptor binding assay using sialylglycopolymers, biolayer interferometry analysis, carbohydrate microarray analysis, crystallography </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> H5N1 infection of human leads to decreased ability to bind 2,3 linked sialic acid </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Matlin, K.S., Reggio, H., Helenius, A. & Simons, K. Infectious entry pathway of influenza-virus in a canine kidney-cell line. J. Cell Biol. 91, 601–613 (1981) </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Electron microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Virus was seen bound to microvilli, in coated pits, coated vesicles, and large smooth-surfaced vacuoles, low pH was required for fusion, suggesting entry by endocytosis </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Rust, M., Lakadamyali, M., Zhang, F. et al. Assembly of endocytic machinery around individual influenza viruses during viral entry. Nat Struct Mol Biol 11, 567–573 (2004). https://doi.org/10.1038/nsmb769 </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Real- time fluorescent microscopy </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Clathrin-mediated and clathrin- and caveolin-independent endocytic pathways used in parallel with similar efficiency </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> De Vries, E. et. al., Dissection of the Influenza A Virus Endocytic Routes Reveals Macropinocytosis as an Alternative Entry Pathway. Plos Pathogens (2011). <a href="https://doi.org/10.1371/journal.ppat.1001329" style="color:#954f72; text-decoration:underline">https://doi.org/10.1371/journal.ppat.1001329</a>   </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Luciferase reporter assay </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> Macropinocytosis is an alternative entry pathway </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Chen, C. and Zhuang, X. Epsin 1 is a cargo- specific adaptor for the clathrin-mediated endocytosis of the influenza virus </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Colocalization of immunofluorescence </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> influenza entry via clathrin- mediated pathway   </td> </tr> <tr> <td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:253px"> Sieczkarski, S. and Whittaker, G. Influenza Virus Can Enter and Infect Cells in the Absence of Clathrin-Mediated Endocytosis </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:190px"> Flow cytommetry </td> <td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:181px"> IAV cell entry via non-clathrin dependent route </td> </tr> </tbody> </table>

-The exact receptor used by IAV for entry remains unknown, and binding is modified by surround glycans as well -Recent findings suggest phsophor-glycans could be a potential alternative receptor -Two subtypes of IAV found exclusively in South and Central American bats (H17N10 and H18N11) use MHC class II for entry

<table class="table table-bordered table-fullwidth"> <thead> <tr> <th>Modulating Factor (MF)</th> <th>MF Specification</th> <th>Effect(s) on the KER</th> <th>Reference(s)</th> </tr> </thead> <tbody> <tr> <td>Viral Origin</td> <td>Viruses originating from different hosts have different preferences (ex: alpha 2,6 vs alpha 2,3 linkage preference)</td> <td>changes tropism, efficiency</td> <td>Rogers GN, Pritchett TJ, Lane JL, Paulson JC. Differential sensitivity of human, avian, and equine influenza A viruses to a glycoprotein inhibitor of infection: selection of receptor specific variants. Virology. 1983 Dec;131(2):394-408. doi: 10.1016/0042-6822(83)90507-x. PMID: 6197808.</td> </tr> </tbody> </table>

Viral infection efficiency follows a Poisson distribution (1). While the rate ans strength of attachment, and speed of entry, have been determined, the exact ratio of binding to entry remains unknown.

The timing from viral attachment to entry has been determined to be 10 minutes (2).

1. Figliozzi RW, Chen F, Chi A, Hsia SC. Using the inverse Poisson distribution to calculate multiplicity of infection and viral replication by a high-throughput fluorescent imaging system. Virol Sin. 2016 Apr;31(2):180-3. doi: 10.1007/s12250-015-3662-8. PMID: 26826079; PMCID: PMC4851903. 2. Dou D, Hernandez-Neuta I, Wang H, Ostbye H, Qian X, Thiele S, et al. Analysis of IAV replication and co-infection dynamics by a versatile RNA viral genome labeling method. Cell Rep (2017) 20:251–63. doi:10.1016/j.celrep.2017.06.021 AOPWiki 2023-08-11T13:42:23

2024-01-18T13:50:21

<upstream-id>aba81357-008c-4064-8eac-a1dd7471917b</upstream-id> <downstream-id>cbc3ed1a-44d2-441b-9e93-e93d02e79ad6</downstream-id>

AOPWiki 2023-08-23T10:48:58

2023-08-23T10:48:58

<upstream-id>cbc3ed1a-44d2-441b-9e93-e93d02e79ad6</upstream-id> <downstream-id>425fd0ff-017c-4593-8a9e-9efb23e32bb8</downstream-id>

AOPWiki 2023-10-03T09:58:42

2023-10-03T09:58:42

<upstream-id>425fd0ff-017c-4593-8a9e-9efb23e32bb8</upstream-id> <downstream-id>7c144eba-aa00-4fef-af25-06d34024fd40</downstream-id>

AOPWiki 2023-10-03T09:58:08

2023-10-03T09:58:08

<upstream-id>7c144eba-aa00-4fef-af25-06d34024fd40</upstream-id> <downstream-id>ca077dd9-2675-4ad3-87d0-3cd4acdabcad</downstream-id>

AOPWiki 2023-10-03T09:59:19

2023-10-03T09:59:19

Binding of Influenza A Virus (IAV) to Sialic Acid Glycan Receptor leads to viral infection proliferation

IAV infection proliferation

Cataia Ives

BY-SA

2.6

This AOP was developed as a proof of concept for the utility of AOPs to model pathogenesis and provide a testable framework for mitigating factor (MF) and countermeasure evaluation. This AOP outlines early key events (KE) leading to viral propagation and spread.

This AOP was developed through focused literature searches of peer-reviewed literature and validated in house (data to be published at a later date).

adjacent

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High adjacent

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Moderate

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Moderate

High

<div> <table class="table table-bordered table-fullwidth"> <thead> <tr> <th>Modulating Factor (MF)</th> <th>Influence or Outcome</th> <th>KER(s) involved</th> </tr> </thead> <tbody> <tr> <td> </td> <td> </td> <td> </td> </tr> </tbody> </table> </div>

Not Specified

AOPWiki 2023-07-31T13:32:46

2024-05-26T20:39:59