Skip to main content
Log in

Bovine viral diarrhea virus structural protein E2 as a complement regulatory protein

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, family Flaviviridae, and is one of the most widely distributed viruses in cattle worldwide. Approximately 60 % of cattle in endemic areas without control measures are infected with BVDV during their lifetime. This wide prevalence of BVDV in cattle populations results in significant economic losses. BVDV is capable of establishing persistent infections in its host due to its ability to infect fetuses, causing immune tolerance. However, this cannot explain how the virus evades the innate immune system. The objective of the present work was to test the potential activity of E2 as a complement regulatory protein. E2 glycoprotein, produced both in soluble and transmembrane forms in stable CHO-K1 cell lines, was able to reduce complement-mediated cell lysis up to 40 % and complement-mediated DNA fragmentation by 50 %, in comparison with cell lines not expressing the glycoprotein. This work provides the first evidence of E2 as a complement regulatory protein and, thus, the finding of a mechanism of immune evasion by BVDV. Furthermore, it is postulated that E2 acts as a self-associated molecular pattern (SAMP), enabling the virus to avoid being targeted by the immune system and to be recognized as self.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Buckwold VE, Beer BE, Donis RO (2003) Bovine viral diarrhea virus as a surrogate model of hepatitis C virus for the evaluation of antiviral agents. Anti Res 60:1–15

    Article  CAS  Google Scholar 

  2. Peterhans E, Bachofen C, Stalder H, Schweizer M (2010) Cytopathic bovine viral diarrhea viruses (BVDV): emerging pestiviruses doomed to extinction. Veterin Res 41:44

    Article  Google Scholar 

  3. Houe H (1999) Epidemiological features and economical importance of bovine virus diarrhoea virus (BVDV) infections. Veterin Microbiol 64:89–107

    Article  CAS  Google Scholar 

  4. Houe H (2003) Economic impact of BVDV infection in dairies. Biologicals 31:137–143

    Article  PubMed  Google Scholar 

  5. Alcami A, Koszinowski UH (2000) Viral mechanisms of immune evasion. Immunol Today 21:447–455

    Article  CAS  PubMed  Google Scholar 

  6. Ploegh HL (1998) Viral strategies of immune evasion. Science 280:248–253

    Article  CAS  PubMed  Google Scholar 

  7. Finlay BB, McFadden G (2006) Anti-immunology: evasion of the host immune system by bacterial and viral pathogens. Cell 124:767–782

    Article  CAS  PubMed  Google Scholar 

  8. Tortorella D, Gewurz BE, Furman MH, Schust DJ, Ploegh HL (2000) Viral subversion of the immune system. Ann Rev Immunol 18:861–926

    Article  CAS  Google Scholar 

  9. Ostachuk A, Chiavenna SM, Gómez C, Pecora A, Pérez-Filgueira MD, Escribano JA, Ardila F, Dus Santos MJ, Wigdorovitz A (2009) Expression of a ScFv-E2T fusion protein in CHO-K1 cells and alfalfa transgenic plants for the selective directioning to antigen presenting cells. Veterin Immunol Immunopathol 128:315

    Article  Google Scholar 

  10. Chiavenna SM, Ostachuk A, Pecora A, Levy S, Dus Santos MJ, Wigdorovitz A (2009) Truncated E2 glicoprotein expression in CHO-K1 cells to produce a subunit vaccine against BVDV. Veterin Immunol Immunopathol 128:231

    Article  Google Scholar 

  11. Escribano JM, Wigdorovitz A, Gil F, Ostachuk A, Pérez-Filgueira M, Domínguez J, Serrano C, Dus Santos MJ, Alonso C (2009) Fusion protein that directs vaccine antigens to antigen presenting cells and applications thereof. World Intellectual Property Organization (WIPO) WO/2009/112603

  12. Ostachuk A, Gil F, Alonso C, Escribano JA, Pérez-Filgueira MD, Wigdorovitz A, Domínguez J, Nuñez C, Dus Santos MJ (2009) Proteína de fusión con direccionamiento de antígenos vacunales a células presentadoras de antígeno y sus aplicaciones. Instituto Nacional de la Propiedad Industrial (INPI) P080101073

  13. Pecora A, Aguirreburualde MSP, Aguirreburualde A, Leunda MR, Odeon A, Chiavenna S, Bochoeyer D, Spitteler M, Filippi J, Dus Santos MJ, Wigdorovitz A (2012) Safety and efficacy of an E2 glycoprotein subunit vaccine produced in mammalian cells to prevent experimental infection with bovine viral diarrhoea virus in cattle. Veterin Res Commun 36:157–164

    Article  Google Scholar 

  14. Aguirreburualde MSP, Gómez MC, Ostachuk A, Wolman F, Albanesi G, Pecora A, Odeon A, Ardila F, Escribano JA, Dus Santos MJ, Wigdorovitz A (2013) Efficacy of a BVDV subunit vaccine produced in alfalfa transgenic plants. Veterin Immunol Immunopathol 151:315–324

    Article  CAS  Google Scholar 

  15. Maurer K, Krey T, Moennig V, Thiel HJ, Rumenapf T (2004) CD46 is a cellular receptor for bovine viral diarrhea virus. J Virol 78:1792–1799

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Krey T, Moussay E, Thiel HJ, Rümenapf T (2006) Role of the low-density lipoprotein receptor in entry of bovine viral diarrhea virus. J Virol 80:10862–10867

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Agnello V, Ábel G, Elfahal M, Knight GB, Zhang QX (1999) Hepatitis C virus and other Flaviviridae viruses enter cells via low density lipoprotein receptor. Proceed Nat Acad Sci 96:12766–12771

    Article  CAS  Google Scholar 

  18. Gladue DP, Baker-Bransetter R, Holinka LG, Fernandez-Sainz IJ, O’Donnell V, Fletcher P, Lu Z, Borca MV (2014) Interaction of CSFV E2 protein with swine host factors as detected by yeast two-hybrid system. PLoS One 9:e85324

    Article  PubMed  PubMed Central  Google Scholar 

  19. Kemper C, Atkinson JP, Hourcade DE (2010) Properdin: emerging roles of a pattern-recognition molecule. Ann Rev Immunol 28:131–155

    Article  CAS  Google Scholar 

  20. Favoreel HW, Van de Walle GR, Nauwynck HJ, Pensaert MB (2003) Virus complement evasion strategies. J General Virol 84:1–15

    Article  CAS  Google Scholar 

  21. Stoermer KA, Morrison TE (2011) Complement and viral pathogenesis. Virology 411:362–373

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bernet J, Mullick J, Singh AK, Sahu A (2003) Viral mimicry of the complement system. J Biosci 28:249–264

    Article  CAS  PubMed  Google Scholar 

  23. Vossen MT, Westerhout EM, Söderberg-Nauclér C, Wiertz EJ (2002) Viral immune evasion: a masterpiece of evolution. Immunogenetics 54:527–542

    Article  CAS  PubMed  Google Scholar 

  24. Meri S, Jördens M, Jarva H (2008) Microbial complement inhibitors as vaccines. Vaccine 26S:I113–I117

    Article  Google Scholar 

  25. Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: SignalP 3.0. J Mole Biol 340:783–795

    Article  Google Scholar 

  26. Hirokawa T, Boon-Chieng S, Mitaku S (1998) SOSUI: classification and secondary structure prediction system for membrane proteins. Bioinformatics 14:378–379

    Article  CAS  PubMed  Google Scholar 

  27. Gupta R, Jung E, Brunak S (2004) Prediction of N-glycosylation sites in human proteins (In preparation)

  28. Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31:3497–3500

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Berg T, King B, Meikle PJ, Nilssen O, Tollersrud OK, Hopwood JJ (2001) Purification and characterization of recombinant human lysosomal [alpha]-mannosidase. Mole Genet Metab 73:18–29

    Article  CAS  Google Scholar 

  30. Goulart HR, dos Santos Arthuso F, Capone MVN, de Oliveira TL, Bartolini P, Soares CRJ (2010) Enhancement of human prolactin synthesis by sodium butyrate addition to serum-free CHO cell culture. J Biomed Biotechnoly 2010:405872

    Google Scholar 

  31. Carvalhal AV, Santos SS, Calado J, Haury M, Carrondo MJT (2003) Cell growth arrest by nucleotides, nucleosides and bases as a tool for improved production of recombinant proteins. Biotechnol Prog 19:69–83

    Article  CAS  PubMed  Google Scholar 

  32. Kumar N, Gammell P, Clynes M (2007) Proliferation control strategies to improve productivity and survival during CHO based production culture. Cytotechnology 53:33–46

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Liu CH, Chen LH (2007) Promotion of recombinant macrophage colony stimulating factor production by dimethyl sulfoxide addition in Chinese hamster ovary cells. J Biosci Bioeng 103:45–49

    Article  CAS  PubMed  Google Scholar 

  34. Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nature Methods 9:671–675

    Article  CAS  PubMed  Google Scholar 

  35. Liszewski MK, Fang CJ, Atkinson JP (2008) Inhibiting complement activation on cells at the step of C3 cleavage. Vaccine 26S:I22–I27

    Article  Google Scholar 

  36. Nauta AJ, Daha MR, Tijsma O, van de Water B, Tedesco F, Roos A (2002) The membrane attack complex of complement induces caspase activation and apoptosis. Euro J Immunol 32:783–792

    Article  Google Scholar 

  37. Cragg M, Howatt W, Bloodworth L, Anderson V, Morgan B, Glennie M (2000) Complement mediated cell death is associated with DNA fragmentation. Cell Death Different 7:48–58

    Article  CAS  Google Scholar 

  38. Elward K, Griffiths M, Mizuno M, Harris CL, Neal JW, Morgan BP, Gasque P (2005) CD46 plays a key role in tailoring innate immune recognition of apoptotic and necrotic cells. J Biol Chem 280:36342–36354

    Article  CAS  PubMed  Google Scholar 

  39. Huang X, Halicka HD, Traganos F, Tanaka T, Kurose A, Darzynkiewicz Z (2005) Cytometric assessment of DNA damage in relation to cell cycle phase and apoptosis. Cell Proliferation 38:223–243

    Article  PubMed  PubMed Central  Google Scholar 

  40. Robinson JP, Rajwa B, Patsekin V, Davisson VJ (2012) Computational analysis of high-throughput flow cytometry data. Expert Opin Drug Dis 7:679–693

    Article  Google Scholar 

  41. Sekiguchi DR, Sutter JA, Rickels MR, Naji A, Liu C, Propert KJ, Rogers WT, Luning Prak ET (2011) Analysis of B cell subsets following pancreatic islet cell transplantation in a patient with type 1 diabetes by cytometric fingerprinting. J Immunol Methods 363:233–244

    Article  CAS  PubMed  Google Scholar 

  42. Rogers WT, Holyst HA (2009) FlowFP: A Bioconductor package for fingerprinting flow cytometric data. Advances in Bioinformatics 2009:193947

    Article  PubMed Central  Google Scholar 

  43. Rogers WT, Moser AR, Holyst HA, Bantly A, Mohler ER III, Scangas G, Moore JS (2008) Cytometric fingerprinting: quantitative characterization of multivariate distributions. Cytometry Part A 73:430–441

    Article  Google Scholar 

  44. Ihaka R, Gentleman R (1996) R: a language for data analysis and graphics. J Comput Graph Stat 5:299–314

    Google Scholar 

  45. Venables WN, Smith DM, R Development Core Team (2002) An introduction to R

  46. Development Core Team R (2008) R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria

    Google Scholar 

  47. Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J et al (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80

    Article  PubMed  PubMed Central  Google Scholar 

  48. Hahne F, LeMeur N, Brinkman R, Ellis B, Haaland P, Sarkar D, Spidlen J, Strain E, Gentleman R (2009) flowCore: a Bioconductor package for high throughput flow cytometry. BMC Bioinform 10:106

    Article  Google Scholar 

  49. Sarkar D, Le Meur N, Gentleman R (2008) Using flowViz to visualize flow cytometry data. Bioinformatics 24:878–879

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Sarkar D (2008) Lattice: multivariate data visualization with R. Springer

  51. Kolde R (2010) Package pheatmap

  52. Tegla C, Cudrici C, Patel S, Trippe R 3rd, Rus V, Niculescu F, Rus H (2011) Membrane attack by complement: the assembly and biology of terminal complement complexes. Immunol Res 51:45–60

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Avirutnan P, Fuchs A, Hauhart RE, Somnuke P, Youn S, Diamond MS, Atkinson JP (2010) Antagonism of the complement component C4 by flavivirus nonstructural protein NS1. J Exp Med 207:793–806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Chung KM, Liszewski MK, Nybakken G, Davis AE, Townsend RR, Fremont DH, Atkinson JP, Diamond MS (2006) West Nile virus nonstructural protein NS1 inhibits complement activation by binding the regulatory protein factor H. Proceed Nat Acad Sci 103:19111–19116

    Article  CAS  Google Scholar 

  55. Dunkelberger JR, Song WC (2010) Complement and its role in innate and adaptive immune responses. Cell Res 20:34–50

    Article  CAS  PubMed  Google Scholar 

  56. Liszewski MK, Post TW, Atkinson JP (1991) Membrane cofactor protein (MCP or CD46): newest member of the regulators of complement activation gene cluster. Ann Rev Immunol 9:431–455

    Article  CAS  Google Scholar 

  57. Van den Berg CW, de la Lastra JMP, Llanes D, Morgan BP (1997) Purification and characterization of the pig analogue of human membrane cofactor protein (CD46/MCP). J Immunol 158:1703–1709

    PubMed  Google Scholar 

  58. Christiansen D, Milland J, Thorley B, McKenzie I, Mottram P, Purcell L, Loveland B (1996) Engineering of recombinant soluble CD46: an inhibitor of complement activation. Immunology 87:348

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Sahu A, Isaacs SN, Soulika AM, Lambris JD (1998) Interaction of vaccinia virus complement control protein with human complement proteins: factor I-mediated degradation of C3b to iC3b1 inactivates the alternative complement pathway. J Immunol 160:5596–5604

    CAS  PubMed  Google Scholar 

  60. Okroj M, Mark L, Stokowska A, Wong SW, Rose N, Blackbourn DJ, Villoutreix BO, Spiller OB, Blom AM (2009) Characterization of the complement inhibitory function of rhesus rhadinovirus complement control protein (RCP). J Biol Chem 284:505–514

    Article  CAS  PubMed  Google Scholar 

  61. Oglesby T, Allen C, Liszewski M, White D, Atkinson J (1992) Membrane cofactor protein (CD46) protects cells from complement-mediated attack by an intrinsic mechanism. J Exp Med 175:1547–1551

    Article  CAS  PubMed  Google Scholar 

  62. Kojima A, Iwata K, Seya T, Matsumoto M, Ariga H, Atkinson JP, Nagasawa S (1993) Membrane cofactor protein (CD46) protects cells predominantly from alternative complement pathway-mediated C3-fragment deposition and cytolysis. J Immunol 151:1519–1527

    CAS  PubMed  Google Scholar 

  63. Riley-Vargas RC, Gill DB, Kemper C, Liszewski MK, Atkinson JP (2004) CD46: expanding beyond complement regulation. Trends Immunol 25:496–503

    Article  CAS  PubMed  Google Scholar 

  64. Liszewski MK, Kemper C, Price JD, Atkinson JP (2005) Emerging roles and new functions of CD46. Springer Seminars in Immunopathology. pp 345–358

  65. Elward K, Gasque P (2003) “Eat me” and “don’t eat me” signals govern the innate immune response and tissue repair in the CNS: emphasis on the critical role of the complement system. Mole Immunol 40:85–94

    Article  CAS  Google Scholar 

  66. Kemper C, Chan AC, Green JM, Brett KA, Murphy KM, Atkinson JP (2003) Activation of human CD4+ cells with CD3 and CD46 induces a T-regulatory cell 1 phenotype. Nature 421:388–392

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

I would like to thank the anonymous reviewers of this manuscript for their thoughtful and constructive suggestions, which led to a substantial improvement of this article. This work was supported by a fellowship from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Agustín Ostachuk.

Ethics declarations

Conflict of interest

The author declares that he has no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ostachuk, A. Bovine viral diarrhea virus structural protein E2 as a complement regulatory protein. Arch Virol 161, 1769–1782 (2016). https://doi.org/10.1007/s00705-016-2835-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-016-2835-6

Keywords

Navigation