1887

Abstract

Equid herpesvirus 1 (EHV-1) causes respiratory disease, abortion and neurological disorders in horses. Cells from the myeloid lineage (CD172a) are one of the main target cells of EHV-1 during primary infection. Recently, we showed that EHV-1 restricts and delays its replication in CD172a cells as part of an immune-evasive strategy to disseminate to target organs. Here, we hypothesize that a low efficiency of EHV-1 binding to and entry in CD172a cells is responsible for this restriction. Thus, we characterized EHV-1 binding and entry into CD172a cells, and showed that EHV-1 only bound to 15–20 % of CD172a cells compared with 70 % of RK-13 control cells. Enzymic removal of heparan sulphate did not reduce EHV-1 infection, suggesting that EHV-1 does not use heparan sulphate to bind and enter CD172a cells. In contrast, we found that treatment of cells with neuraminidase (NA) reduced infection by 85–100 % compared with untreated cells, whilst NA treatment of virus had no effect on infection. This shows that sialic acid residues present on CD172a cells are essential in the initiation of EHV-1 infection. We found that αβ integrins are involved in the post-binding stage of CD172a cell infection. Using pharmacological inhibitors, we showed that EHV-1 does not enter CD172a cells via a clathrin- or caveolae-dependent endocytic pathway, nor by macropinocytosis, but requires cholesterol, tyrosine kinase, actin, dynamin and endosomal acidification, pointing towards a phagocytic mechanism. Overall, these results show that the narrow tropism of EHV-1 amongst CD172a cells is determined by the presence of specific cellular receptors.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000375
2016-03-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/97/3/733.html?itemId=/content/journal/jgv/10.1099/jgv.0.000375&mimeType=html&fmt=ahah

References

  1. Allen G. P., Bryans J. T. 1986; Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections. Prog Vet Microbiol Immunol 2:78–144[PubMed]
    [Google Scholar]
  2. Andoh K., Hattori S., Mahmoud H. Y., Takasugi M., Shimoda H., Bannai H., Tsujimura K., Matsumura T., Kondo T., other authors. 2015; The haemagglutination activity of equine herpesvirus type 1 glycoprotein C. Virus Res 195:172–176 [View Article][PubMed]
    [Google Scholar]
  3. Atanasiu D., Whitbeck J. C., Cairns T. M., Reilly B., Cohen G. H., Eisenberg R. J. 2007; Bimolecular complementation reveals that glycoproteins gB and gH/gL of herpes simplex virus interact with each other during cell fusion. Proc Natl Acad Sci U S A 104:18718–18723 [View Article][PubMed]
    [Google Scholar]
  4. Azab W., Osterrieder N. 2012; Glycoproteins D of equine herpesvirus type 1 (EHV-1) and EHV-4 determine cellular tropism independently of integrins. J Virol 86:2031–2044 [View Article][PubMed]
    [Google Scholar]
  5. Barretto N., Hallak L. K., Peeples M. E. 2003; Neuraminidase treatment of respiratory syncytial virus-infected cells or virions, but not target cells, enhances cell-cell fusion and infection. Virology 313:33–43 [View Article][PubMed]
    [Google Scholar]
  6. Cahan L. D., Singh R., Paulson J. C. 1983; Sialyloligosaccharide receptors of binding variants of polyoma virus. Virology 130:281–289 [View Article][PubMed]
    [Google Scholar]
  7. Chesnokova L. S., Nishimura S. L., Hutt-Fletcher L. M. 2009; Fusion of epithelial cells by Epstein-Barr virus proteins is triggered by binding of viral glycoproteins gHgL to integrins alphavbeta6 or alphavbeta8. Proc Natl Acad Sci U S A 106:20464–20469 [View Article][PubMed]
    [Google Scholar]
  8. Clement C., Tiwari V., Scanlan P. M., Valyi-Nagy T., Yue B. Y., Shukla D. 2006; A novel role for phagocytosis-like uptake in herpes simplex virus entry. J Cell Biol 174:1009–1021 [View Article][PubMed]
    [Google Scholar]
  9. Compton T., Nowlin D. M., Cooper N. R. 1993; Initiation of human cytomegalovirus infection requires initial interaction with cell surface heparan sulfate. Virology 193:834–841 [View Article][PubMed]
    [Google Scholar]
  10. Connor R. J., Kawaoka Y., Webster R. G., Paulson J. C. 1994; Receptor specificity in human, avian, and equine H2 and H3 influenza virus isolates. Virology 205:17–23 [View Article][PubMed]
    [Google Scholar]
  11. Delputte P. L., Nauwynck H. J. 2004; Porcine arterivirus infection of alveolar macrophages is mediated by sialic acid on the virus. J Virol 78:8094–8101 [View Article][PubMed]
    [Google Scholar]
  12. Di A., Nelson D. J., Bindokas V., Brown M. E., Libunao F., Palfrey H. C. 2003; Dynamin regulates focal exocytosis in phagocytosing macrophages. Mol Biol Cell 14:2016–2028 [View Article][PubMed]
    [Google Scholar]
  13. El-Sayed A., Harashima H. 2013; Endocytosis of gene delivery vectors: from clathrin-dependent to lipid raft-mediated endocytosis. Mol Ther 21:1118–1130 [View Article][PubMed]
    [Google Scholar]
  14. Flannagan R. S., Jaumouillé V., Grinstein S. 2012; The cell biology of phagocytosis. Annu Rev Pathol 7:61–98 [View Article][PubMed]
    [Google Scholar]
  15. Frampton A. R. Jr., Stolz D. B., Uchida H., Goins W. F., Cohen J. B., Glorioso J. C. 2007; Equine herpesvirus 1 enters cells by two different pathways, and infection requires the activation of the cellular kinase ROCK1. J Virol 81:10879–10889 [View Article][PubMed]
    [Google Scholar]
  16. Gamblin S. J., Haire L. F., Russell R. J., Stevens D. J., Xiao B., Ha Y., Vasisht N., Steinhauer D. A., Daniels R. S., other authors. 2004; The structure and receptor binding properties of the 1918 influenza hemagglutinin. Science 303:1838–1842 [View Article][PubMed]
    [Google Scholar]
  17. Garré B., Gryspeerdt A., Croubels S., De Backer P., Nauwynck H. 2009; Evaluation of orally administered valacyclovir in experimentally EHV1-infected ponies. Vet Microbiol 135:214–221 [View Article][PubMed]
    [Google Scholar]
  18. Garrigues H. J., Rubinchikova Y. E., Dipersio C. M., Rose T. M. 2008; Integrin alphaVbeta3 binds to the RGD motif of glycoprotein B of Kaposi's sarcoma-associated herpesvirus and functions as an RGD-dependent entry receptor. J Virol 82:1570–1580 [View Article][PubMed]
    [Google Scholar]
  19. Garrigues H. J., DeMaster L. K., Rubinchikova Y. E., Rose T. M. 2014; KSHV attachment and entry are dependent on αVβ3 integrin localized to specific cell surface microdomains and do not correlate with the presence of heparan sulfate. Virology 464-465:118–133 [View Article][PubMed]
    [Google Scholar]
  20. Gianni T., Gatta V., Campadelli-Fiume G. 2010; alphaVbeta3-integrin routes herpes simplex virus to an entry pathway dependent on cholesterol-rich lipid rafts and dynamin2. Proc Natl Acad Sci U S A 107:22260–22265 [View Article][PubMed]
    [Google Scholar]
  21. Gold E. S., Underhill D. M., Morrissette N. S., Guo J., McNiven M. A., Aderem A. 1999; Dynamin 2 is required for phagocytosis in macrophages. J Exp Med 190:1849–1856 [View Article][PubMed]
    [Google Scholar]
  22. Goodenow M. M., Collman R. G. 2006; HIV-1 coreceptor preference is distinct from target cell tropism: a dual-parameter nomenclature to define viral phenotypes. J Leukoc Biol 80:965–972 [View Article][PubMed]
    [Google Scholar]
  23. Gryspeerdt A. C., Vandekerckhove A. P., Baghi H. B., van de Walle G. R., Nauwynck H. J. 2012; Expression of late viral proteins is restricted in nasal mucosal leucocytes but not in epithelial cells during early-stage equine herpes virus-1 infection. Vet J 193:576–578 [View Article][PubMed]
    [Google Scholar]
  24. Hasebe R., Sasaki M., Sawa H., Wada R., Umemura T., Kimura T. 2009; Infectious entry of equine herpesvirus-1 into host cells through different endocytic pathways. Virology 393:198–209 [View Article][PubMed]
    [Google Scholar]
  25. Kannan S., Audet A., Huang H., Chen L. J., Wu M. 2008; Cholesterol-rich membrane rafts and Lyn are involved in phagocytosis during Pseudomonas aeruginosa infection. J Immunol 180:2396–2408 [View Article][PubMed]
    [Google Scholar]
  26. Kurtz B. M., Singletary L. B., Kelly S. D., Frampton A.R., Jr.. 2010; Equus caballus major histocompatibility complex class I is an entry receptor for equine herpesvirus type 1. J Virol 84:9027–9034 [View Article][PubMed]
    [Google Scholar]
  27. Laval K., Favoreel H. W., Nauwynck H. J. 2015a; Equine herpesvirus type 1 replication is delayed in CD172a+ monocytic cells and controlled by histone deacetylases. J Gen Virol 96:118–130 [View Article][PubMed]
    [Google Scholar]
  28. Laval K., Favoreel H. W., Poelaert K. C., Van Cleemput J., Nauwynck H. J. 2015b; Equine herpesvirus type 1 enhances viral replication in CD172a+ monocytic cells upon adhesion to endothelial cells. J Virol 89:10912–10923 [View Article][PubMed]
    [Google Scholar]
  29. Macia E., Ehrlich M., Massol R., Boucrot E., Brunner C., Kirchhausen T. 2006; Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell 10:839–850 [View Article][PubMed]
    [Google Scholar]
  30. Marsh M., Helenius A. 2006; Virus entry: open sesame. Cell 124:729–740 [View Article][PubMed]
    [Google Scholar]
  31. Matrosovich M., Tuzikov A., Bovin N., Gambaryan A., Klimov A., Castrucci M. R., Donatelli I., Kawaoka Y. 2000; Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals. J Virol 74:8502–8512 [View Article][PubMed]
    [Google Scholar]
  32. Matrosovich M., Herrler G., Klenk H. D. 2015; Sialic acid receptors of viruses. Top Curr Chem 367:1–28 [View Article][PubMed]
    [Google Scholar]
  33. Mercer J., Helenius A. 2009; Virus entry by macropinocytosis. Nat Cell Biol 11:510–520 [View Article][PubMed]
    [Google Scholar]
  34. Mettlen M., Pucadyil T., Ramachandran R., Schmid S. L. 2009; Dissecting dynamin's role in clathrin-mediated endocytosis. Biochem Soc Trans 37:1022–1026 [View Article][PubMed]
    [Google Scholar]
  35. Nabi I. R., Le P. U. 2003; Caveolae/raft-dependent endocytosis. J Cell Biol 161:673–677 [View Article][PubMed]
    [Google Scholar]
  36. Neubauer A., Braun B., Brandmuller C., Kaaden O. R., Osterrieder N. 1997; Analysis of the contributions of the equine herpesvirus 1 glycoprotein gB homolog to virus entry and direct cell-to-cell spread. Virology 227:281–294 [View Article][PubMed]
    [Google Scholar]
  37. Nicola A. V., McEvoy A. M., Straus S. E. 2003; Roles for endocytosis and low pH in herpes simplex virus entry into HeLa and Chinese hamster ovary cells. J Virol 77:5324–5332 [View Article][PubMed]
    [Google Scholar]
  38. Nicola A. V., Hou J., Major E. O., Straus S. E. 2005; Herpes simplex virus type 1 enters human epidermal keratinocytes, but not neurons, via a pH-dependent endocytic pathway. J Virol 79:7609–7616 [View Article][PubMed]
    [Google Scholar]
  39. Osterrieder N. 1999; Construction and characterization of an equine herpesvirus 1 glycoprotein C negative mutant. Virus Res 59:165–177 [View Article][PubMed]
    [Google Scholar]
  40. Patel J. R., Heldens J. 2005; Equine herpesviruses 1 (EHV-1) and 4 (EHV-4) - epidemiology, disease and immunoprophylaxis: a brief review. Vet J 170:14–23 [View Article][PubMed]
    [Google Scholar]
  41. Pelkmans L. 2005; Secrets of caveolae- and lipid raft-mediated endocytosis revealed by mammalian viruses. Biochim Biophys Acta 1746:295–304 [View Article][PubMed]
    [Google Scholar]
  42. Pelkmans L., Helenius A. 2003; Insider information: what viruses tell us about endocytosis. Curr Opin Cell Biol 15:414–422 [View Article][PubMed]
    [Google Scholar]
  43. Sasaki M., Hasebe R., Makino Y., Suzuki T., Fukushi H., Okamoto M., Matsuda K., Taniyama H., Sawa H., Kimura T. 2011; Equine major histocompatibility complex class I molecules act as entry receptors that bind to equine herpesvirus-1 glycoprotein D. Genes Cells 16:343–357 [View Article][PubMed]
    [Google Scholar]
  44. Shieh M. T., WuDunn D., Montgomery R. I., Esko J. D., Spear P. G. 1992; Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J Cell Biol 116:1273–1281 [View Article][PubMed]
    [Google Scholar]
  45. Shukla D., Spear P. G. 2001; Herpesviruses and heparan sulfate: an intimate relationship in aid of viral entry. J Clin Invest 108:503–510 [View Article][PubMed]
    [Google Scholar]
  46. Skehel J. J., Wiley D. C. 2000; Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. Annu Rev Biochem 69:531–569 [View Article][PubMed]
    [Google Scholar]
  47. Teuton J. R., Brandt C. R. 2007; Sialic acid on herpes simplex virus type 1 envelope glycoproteins is required for efficient infection of cells. J Virol 81:3731–3739 [View Article][PubMed]
    [Google Scholar]
  48. Trybala E., Bergström T., Spillmann D., Svennerholm B., Flynn S. J., Ryan P. 1998; Interaction between pseudorabies virus and heparin/heparan sulfate. Pseudorabies virus mutants differ in their interaction with heparin/heparan sulfate when altered for specific glycoprotein C heparin-binding domain. J Biol Chem 273:5047–5052 [View Article][PubMed]
    [Google Scholar]
  49. van de Walle G. R., Peters S. T., VanderVen B. C., O'Callaghan D. J., Osterrieder N. 2008; Equine herpesvirus 1 entry via endocytosis is facilitated by alphaV integrins and an RSD motif in glycoprotein D. J Virol 82:11859–11868 [View Article][PubMed]
    [Google Scholar]
  50. van der Meulen K. M., Nauwynck H. J., Buddaert W., Pensaert M. B. 2000; Replication of equine herpesvirus type 1 in freshly isolated equine peripheral blood mononuclear cells and changes in susceptibility following mitogen stimulation. J Gen Virol 81:21–25 [View Article][PubMed]
    [Google Scholar]
  51. van der Meulen K. M., Vercauteren G., Nauwynck H. J. 2003a; A local epidemic of equine herpesvirus 1-induced neurological disorders in Belgium. Vlaams Diergen Tijds 72:186–194
    [Google Scholar]
  52. Wang X., Huang D. Y., Huong S. M., Huang E. S. 2005; Integrin alphavbeta3 is a coreceptor for human cytomegalovirus. Nat Med 11:515–521 [View Article][PubMed]
    [Google Scholar]
  53. Wittels M., Spear P. G. 1991; Penetration of cells by herpes simplex virus does not require a low pH-dependent endocytic pathway. Virus Res 18:271–290 [View Article][PubMed]
    [Google Scholar]
  54. Wu W., Air G. M. 2004; Binding of influenza viruses to sialic acids: reassortant viruses with A/NWS/33 hemagglutinin bind to α2,8-linked sialic acid. Virology 325:340–350 [View Article][PubMed]
    [Google Scholar]
  55. Yang X., Steukers L., Forier K., Xiong R., Braeckmans K., Van Reeth K., Nauwynck H. 2014; A beneficiary role for neuraminidase in influenza virus penetration through the respiratory mucus. PLoS One 9:e110026 [View Article][PubMed]
    [Google Scholar]
  56. Zou Z., Chastain A., Moir S., Ford J., Trandem K., Martinelli E., Cicala C., Crocker P., Arthos J., Sun P. D. 2011; Siglecs facilitate HIV-1 infection of macrophages through adhesion with viral sialic acids. PLoS One 6:e24559 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000375
Loading
/content/journal/jgv/10.1099/jgv.0.000375
Loading

Data & Media loading...

Supplements

Supplementary Data

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error