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Journal of General Virology header logo Journal of General Virology

Volume 106, Issue 6

agglutinin enhances entry through interactions at specific -linked glycosylation sites on the virus glycoprotein complex Open Access

Abstract

Entry of e (EBOV) into a host cell is a complex process requiring interactions between the viral glycoproteins (GPs) and cellular factors. These entry factors are cell-specific and can include cell surface lectins and phosphatidylserine receptors. Niemann–Pick type C1 is critical to the late stage of the entry process. Entry has been demonstrated to be enhanced by interactions between the virion and surface-expressed lectins, which interact with carbohydrate moieties attached to the GP. In addition, soluble lectins, including mannose-binding lectin, can enhance entry . However, the mechanism of lectin-mediated enhancement remains to be defined. This study investigated the possibility that plant lectins, agglutinin (WFA), soybean agglutinin (SBA) and agglutinin (GNA), which possess different carbohydrate-binding specificities, influence EBOV entry. WFA was observed to potently enhance entry of lentiviral pseudotype viruses (PVs) expressing the GP of three species [Zaire, Sudan () and Reston ()], with the greatest impact on EBOV. SBA had a modest enhancing effect on entry that was specific to EBOV, whilst GNA had no impact on the entry of any of the species. None of the lectins enhanced the entry of control PVs expressing the surface proteins of other RNA viruses tested. WFA was demonstrated to bind directly with the EBOV-GP via the glycans, and mutational analysis implicated N as contributing to the interaction. Furthermore, enhancement was observed in both human and bat cell lines, indicating a highly conserved mechanism of action. We conclude that the binding of WFA to EBOV-GP through interactions including the glycan at N results in GP alterations that enhance entry, providing evidence of a mechanism for lectin-mediated virus entry enhancement. Targeting lectin-ligand interactions presents a potential strategy for restricting entry.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Ebola virus , enhancement , lectin , pseudotype and virus entry
Funding
This study was supported by the:
  • Medical Research Council (Award MR/S009434/1)
    • Principle Award Recipient: JonathanK Ball
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2025-06-06
2025-06-24
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References

  1. Letafati A, Salahi Ardekani O, Karami H, Soleimani M. Ebola virus disease: a narrative review. Microb Pathog 2023; 181:106213 [View Article] [PubMed]
     [Google Scholar]
  2. Feldmann H, Geisbert TW. Ebola haemorrhagic fever. Lancet 2011; 377:849–862 [View Article] [PubMed]
     [Google Scholar]
  3. Aruna A, Mbala P, Minikulu L, Mukadi D, Bulemfu D et al. Ebola virus disease outbreak - Democratic Republic of the Congo, August 2018-November 2019. MMWR Morb Mortal Wkly Rep 2019; 68:1162–1165 [View Article] [PubMed]
     [Google Scholar]
  4. Kyobe Bosa H, Kamara N, Aragaw M, Wayengera M, Talisuna A et al. The West Africa Ebola virus disease outbreak: 10 years on. Lancet Glob Heal 2024; 12:e1081–e1083 [View Article]
     [Google Scholar]
  5. Feldmann H, Volchkov VE, Volchkova VA, Ströher U, Klenk H-D. Biosynthesis and role of filoviral glycoproteins. J Gen Virol 2001; 82:2839–2848 [View Article] [PubMed]
     [Google Scholar]
  6. Lee JE, Fusco ML, Hessell AJ, Oswald WB, Burton DR et al. Structure of the Ebola virus glycoprotein bound to an antibody from a human survivor. Nature 2008; 454:177–182 [View Article] [PubMed]
     [Google Scholar]
  7. Spence JS, Krause TB, Mittler E, Jangra RK, Chandran K. Direct visualization of Ebola virus fusion triggering in the endocytic pathway. mBio 2016; 7:e01857-15 [View Article] [PubMed]
     [Google Scholar]
  8. Nathan L, Lai AL, Millet JK, Straus MR, Freed JH et al. Calcium ions directly interact with the Ebola virus fusion peptide to promote structure-function changes that enhance infection. ACS Infect Dis 2020; 6:250–260 [View Article] [PubMed]
     [Google Scholar]
  9. Chandran K, Sullivan NJ, Felbor U, Whelan SP, Cunningham JM. Endosomal proteolysis of the Ebola virus glycoprotein is necessary for infection. Science 2005; 308:1643–1645 [View Article] [PubMed]
     [Google Scholar]
  10. Carette JE, Raaben M, Wong AC, Herbert AS, Obernosterer G et al. Ebola virus entry requires the cholesterol transporter Niemann-Pick C1. Nature 2011; 477:340–343 [View Article] [PubMed]
     [Google Scholar]
  11. Bornholdt ZA, Ndungo E, Fusco ML, Bale S, Flyak AI et al. Host-primed Ebola virus GP exposes a hydrophobic NPC1 receptor-binding pocket, revealing a target for broadly neutralizing antibodies. mBio 2016; 7:e02154–15 [View Article] [PubMed]
     [Google Scholar]
  12. Iraqi M, Edri A, Greenshpan Y, Kundu K, Bolel P et al. N-glycans mediate the Ebola virus-GP1 shielding of ligands to immune receptors and immune evasion. Front Cell Infect Microbiol 2020; 10:48 [View Article] [PubMed]
     [Google Scholar]
  13. Peng W, Rayaprolu V, Parvate AD, Pronker MF, Hui S et al. Glycan shield of the ebolavirus envelope glycoprotein GP. Commun Biol 2022; 5:785 [View Article] [PubMed]
     [Google Scholar]
  14. Takada A, Robison C, Goto H, Sanchez A, Murti KG et al. A system for functional analysis of Ebola virus glycoprotein. Proc Natl Acad Sci USA 1997; 94:14764–14769 [View Article] [PubMed]
     [Google Scholar]
  15. Alvarez CP, Lasala F, Carrillo J, Muñiz O, Corbí AL et al. C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans. J Virol 2002; 76:6841–6844 [View Article] [PubMed]
     [Google Scholar]
  16. Simmons G, Reeves JD, Grogan CC, Vandenberghe LH, Baribaud F et al. DC-SIGN and DC-SIGNR bind ebola glycoproteins and enhance infection of macrophages and endothelial cells. Virology 2003; 305:115–123 [View Article] [PubMed]
     [Google Scholar]
  17. Lepenies B, Lang R. Editorial: lectins and their ligands in shaping immune responses. Front Immunol 2019; 10:2379 [View Article] [PubMed]
     [Google Scholar]
  18. Mason CP, Tarr AW. Human lectins and their roles in viral infections. Molecules 2015; 20:2229–2271 [View Article] [PubMed]
     [Google Scholar]
  19. Zhang W, Bouwman KM, van Beurden SJ, Ordonez SR, van Eijk M et al. Chicken mannose binding lectin has antiviral activity towards infectious bronchitis virus. Virology 2017; 509:252–259 [View Article] [PubMed]
     [Google Scholar]
  20. Verma A, White M, Vathipadiekal V, Tripathi S, Mbianda J et al. Human H-ficolin inhibits replication of seasonal and pandemic influenza A viruses. J Immunol 2012; 189:2478–2487 [View Article] [PubMed]
     [Google Scholar]
  21. Avirutnan P, Hauhart RE, Marovich MA, Garred P, Atkinson JP et al. Complement-mediated neutralization of dengue virus requires mannose-binding lectin. mBio 2011; 2:1–11 [View Article] [PubMed]
     [Google Scholar]
  22. Jalal PJ, Urbanowicz RA, Horncastle E, Pathak M, Goddard C et al. Expression of human ficolin-2 in hepatocytes confers resistance to infection by diverse hepatotropic viruses. J Med Microbiol 2019; 68:642–648 [View Article] [PubMed]
     [Google Scholar]
  23. Brown KS, Keogh MJ, Owsianka AM, Adair R, Patel AH et al. Specific interaction of hepatitis C virus glycoproteins with mannan binding lectin inhibits virus entry. Protein Cell 2010; 1:664–674 [View Article] [PubMed]
     [Google Scholar]
  24. Hamed MR, Brown RJP, Zothner C, Urbanowicz RA, Mason CP et al. Recombinant human L-ficolin directly neutralizes hepatitis C virus entry. J Innate Immun 2014; 6:676–684 [View Article] [PubMed]
     [Google Scholar]
  25. Lin G, Simmons G, Pöhlmann S, Baribaud F, Ni H et al. Differential N-linked glycosylation of human immunodeficiency virus and Ebola virus envelope glycoproteins modulates interactions with DC-SIGN and DC-SIGNR. J Virol 2003; 77:1337–1346 [View Article] [PubMed]
     [Google Scholar]
  26. Takada A, Fujioka K, Tsuiji M, Morikawa A, Higashi N et al. Human macrophage C-type lectin specific for galactose and N-acetylgalactosamine promotes filovirus entry. J Virol 2004; 78:2943–2947 [View Article] [PubMed]
     [Google Scholar]
  27. Messingham KN, Richards PT, Fleck A, Patel RA, Djurkovic M et al. Multiple cell types support productive infection and dynamic translocation of infectious Ebola virus to the surface of human skin. Sci Adv 2025; 11:eadr6140 [View Article] [PubMed]
     [Google Scholar]
  28. Marzi A, Möller P, Hanna SL, Harrer T, Eisemann J et al. Analysis of the interaction of Ebola virus glycoprotein with DC-SIGN (dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin) and its homologue DC-SIGNR. J Infect Dis 2007; 196 Suppl 2:S237–46 [View Article] [PubMed]
     [Google Scholar]
  29. Ji X, Olinger GG, Aris S, Chen Y, Gewurz H et al. Mannose-binding lectin binds to Ebola and Marburg envelope glycoproteins, resulting in blocking of virus interaction with DC-SIGN and complement-mediated virus neutralization. J Gen Virol 2005; 86:2535–2542 [View Article] [PubMed]
     [Google Scholar]
  30. Michelow IC, Lear C, Scully C, Prugar LI, Longley CB et al. High-dose mannose-binding lectin therapy for Ebola virus infection. J Infect Dis 2011; 203:175–179 [View Article] [PubMed]
     [Google Scholar]
  31. Brudner M, Karpel M, Lear C, Chen L, Yantosca LM et al. Lectin-dependent enhancement of Ebola virus infection via soluble and transmembrane C-type lectin receptors. PLoS One 2013; 8:e60838 [View Article] [PubMed]
     [Google Scholar]
  32. Favier A-L, Gout E, Reynard O, Ferraris O, Kleman J-P et al. Enhancement of Ebola virus infection via ficolin-1 interaction with the mucin domain of GP glycoprotein. J Virol 2016; 90:5256–5269 [View Article] [PubMed]
     [Google Scholar]
  33. Tsaneva M, Van Damme EJM. 130 years of plant lectin research. Glycoconj J 2020; 37:533–551 [View Article] [PubMed]
     [Google Scholar]
  34. Mazalovska M, Kouokam JC. Lectins as promising therapeutics for the prevention and treatment of HIV and other potential coinfections. Biomed Res Int 2018; 2018:3750646 [View Article] [PubMed]
     [Google Scholar]
  35. Marchetti M, Mastromarino P, Rieti S, Seganti L, Orsi N. Inhibition of herpes simplex, rabies and rubella viruses by lectins with different specificities. Res Virol 1995; 146:211–215 [View Article] [PubMed]
     [Google Scholar]
  36. Swanson MD, Boudreaux DM, Salmon L, Chugh J, Winter HC et al. Engineering a therapeutic lectin by uncoupling mitogenicity from antiviral activity. Cell 2015; 163:746–758 [View Article] [PubMed]
     [Google Scholar]
  37. Covés-Datson EM, Dyall J, DeWald LE, King SR, Dube D et al. Inhibition of Ebola virus by a molecularly engineered banana lectin. PLoS Negl Trop Dis 2019; 13:e0007595 [View Article] [PubMed]
     [Google Scholar]
  38. Sanchez A, Yang ZY, Xu L, Nabel GJ, Crews T et al. Biochemical analysis of the secreted and virion glycoproteins of Ebola virus. J Virol 1998; 72:6442–6447 [View Article] [PubMed]
     [Google Scholar]
  39. Haji-Ghassemi O, Gilbert M, Spence J, Schur MJ, Parker MJ et al. Molecular basis for recognition of the cancer glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. J Biol Chem 2016; 291:24085–24095 [View Article] [PubMed]
     [Google Scholar]
  40. Gupta D, Bhattacharyya L, Fant J et al. Observation of unique cross-linked lattices between multiantennary carbohydrates and soybean lectin. presence of pseudo-2-fold axes of symmetry in complex type carbohydrates. Biochemistry 1994; 33: [View Article]
     [Google Scholar]
  41. Shibuya N, Goldstein IJ, Van Damme EJ, Peumans WJ. Binding properties of a mannose-specific lectin from the snowdrop (Galanthus nivalis) bulb. J Biol Chem 1988; 263:728–734 [View Article] [PubMed]
     [Google Scholar]
  42. Urbanowicz RA, McClure CP, Sakuntabhai A, Sall AA, Kobinger G et al. Human adaptation of Ebola virus during the West African outbreak. Cell 2016; 167:1079–1087 [View Article] [PubMed]
     [Google Scholar]
  43. Urbanowicz RA, McClure CP, King B, Mason CP, Ball JK et al. Novel functional hepatitis C virus glycoprotein isolates identified using an optimized viral pseudotype entry assay. J Gen Virol 2016; 97:2265–2279 [View Article] [PubMed]
     [Google Scholar]
  44. Nakabayashi H, Taketa K, Miyano K, Yamane T, Sato J. Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res 1982; 42:3858–3863 [PubMed]
     [Google Scholar]
  45. Metheringham RL, Pudney VA, Gunn B, Towey M, Spendlove I et al. Antibodies designed as effective cancer vaccines. MAbs 2009; 1:71–85 [View Article] [PubMed]
     [Google Scholar]
  46. Augestad EH, Holmboe Olesen C, Grønberg C, Soerensen A, Velázquez-Moctezuma R et al. The hepatitis C virus envelope protein complex is a dimer of heterodimers. Nature 2024; 633:704–709 [View Article] [PubMed]
     [Google Scholar]
  47. Shaw J, Gosain R, Kalita MM, Foster TL, Kankanala J et al. Rationally derived inhibitors of hepatitis C virus (HCV) p7 channel activity reveal prospect for bimodal antiviral therapy. elife 2020; 9:1–36 [View Article] [PubMed]
     [Google Scholar]
  48. Ashfaq UA, Masoud MS, Khaliq S, Nawaz Z, Riazuddin S. Inhibition of hepatitis C virus 3a genotype entry through Glanthus Nivalis Agglutinin. Virol J 2011; 8:248 [View Article] [PubMed]
     [Google Scholar]
  49. Kühl A, Hoffmann M, Müller MA, Munster VJ, Gnirss K et al. Comparative analysis of Ebola virus glycoprotein interactions with human and bat cells. J Infect Dis 2011; 204 Suppl 3:S840–9 [View Article] [PubMed]
     [Google Scholar]
  50. Leroy EM, Kumulungui B, Pourrut X, Rouquet P, Hassanin A et al. Fruit bats as reservoirs of Ebola virus. Nature 2005; 438:575–576 [View Article] [PubMed]
     [Google Scholar]
  51. Diehl WE, Lin AE, Grubaugh ND, Carvalho LM, Kim K et al. Ebola virus glycoprotein with increased infectivity dominated the 2013–2016 epidemic. Cell 2016; 167:1088–1098 [View Article] [PubMed]
     [Google Scholar]
  52. Wang MK, Lim SY, Lee SM, Cunningham JM. Biochemical basis for increased activity of Ebola glycoprotein in the 2013–16 epidemic. Cell Host Microbe 2017; 21:367–375 [View Article] [PubMed]
     [Google Scholar]
  53. Fels JM, Bortz RH 3rd, Alkutkar T, Mittler E, Jangra RK et al. A glycoprotein mutation that emerged during the 2013–2016 Ebola virus epidemic alters proteolysis and accelerates membrane fusion. mBio 2021; 12:03616–03620 [View Article] [PubMed]
     [Google Scholar]
  54. Lennemann NJ, Rhein BA, Ndungo E, Chandran K, Qiu X et al. Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. mBio 2014; 5:e00862-13 [View Article] [PubMed]
     [Google Scholar]
  55. Collar AL, Clarke EC, Anaya E, Merrill D, Yarborough S et al. Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. Virology 2017; 502:39–47 [View Article] [PubMed]
     [Google Scholar]
  56. Ng M, Ndungo E, Kaczmarek ME, Herbert AS, Binger T et al. Filovirus receptor NPC1 contributes to species-specific patterns of ebolavirus susceptibility in bats. elife 2015; 4:1–22 [View Article] [PubMed]
     [Google Scholar]
  57. Wang H, Shi Y, Song J, Qi J, Lu G et al. Ebola viral glycoprotein bound to its endosomal receptor Niemann-Pick C1. Cell 2016; 164:258–268 [View Article] [PubMed]
     [Google Scholar]
  58. Steeds K, Hall Y, Slack GS, Longet S, Strecker T et al. Pseudotyping of VSV with Ebola virus glycoprotein is superior to HIV-1 for the assessment of neutralising antibodies. Sci Rep 2020; 10:14289 [View Article] [PubMed]
     [Google Scholar]
  59. Cantoni D, Wilkie C, Bentley EM, Mayora-Neto M, Wright E et al. Correlation between pseudotyped virus and authentic virus neutralisation assays, a systematic review and meta-analysis of the literature. Front Immunol 2023; 14:1184362 [View Article] [PubMed]
     [Google Scholar]
  60. Watanabe S, Watanabe T, Noda T, Takada A, Feldmann H et al. Production of novel ebola virus-like particles from cDNAs: an alternative to ebola virus generation by reverse genetics. J Virol 2004; 78:999–1005 [View Article] [PubMed]
     [Google Scholar]
  61. Wang B, Wang Y, Frabutt DA, Zhang X, Yao X et al. Mechanistic understanding of N-glycosylation in Ebola virus glycoprotein maturation and function. J Biol Chem 2017; 292:5860–5870 [View Article]
     [Google Scholar]
  62. Jeffers SA, Sanders DA, Sanchez A. Covalent modifications of the ebola virus glycoprotein. J Virol 2002; 76:12463–12472 [View Article] [PubMed]
     [Google Scholar]
  63. Uhlén M, Fagerberg L, Hallström BM, Lindskog C, Oksvold P et al. Tissue-based map of the human proteome. Science 2015; 347:1260419 [View Article] [PubMed]
     [Google Scholar]
  64. Jin H, Zhang C, Zwahlen M, von Feilitzen K, Karlsson M et al. Systematic transcriptional analysis of human cell lines for gene expression landscape and tumor representation. Nat Commun 2023; 14:5417 [View Article] [PubMed]
     [Google Scholar]
  65. Matsuno K, Nakayama E, Noyori O, Marzi A, Ebihara H et al. C-type lectins do not act as functional receptors for filovirus entry into cells. Biochem Biophys Res Commun 2010; 403:144–148 [View Article] [PubMed]
     [Google Scholar]
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Wisteria floribunda agglutinin enhances Zaire ebolavirus entry through interactions at specific N-linked glycosylation sites on the virus glycoprotein complex
J. Gen. Virol. 106, 002120 (2025); https://doi.org/10.1099/jgv.0.002120
/content/journal/jgv/10.1099/jgv.0.002120
/content/journal/jgv/10.1099/jgv.0.002120
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