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An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

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Abstract

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6–8 kb.

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References

  1. Breitburd F, Kirnbauer R, Hubbert NL, Nonnenmacher B, Trin DD, Orth G, Schiller JT, Lowy DR (1995) Immunization with viruslike particles from cottontail rabbit papillomavirus (CRPV) can protect against experimental CRPV infection. J Virol 69:3959–3963

    PubMed  CAS  Google Scholar 

  2. Finnen RL, Erickson KD, Chen XS, Garcea RL (2003) Interactions between papillomavirus L1 and L2 capsid proteins. J Virol 77:4818–4826

    Article  PubMed  CAS  Google Scholar 

  3. Frazer IH (2004) Prevention of cervical cancer through papillomavirus vaccination. Nat Rev Immunol 4:46–55

    Article  PubMed  CAS  Google Scholar 

  4. Greenstone HL, Nieland JD, de Visser KE, De Bruijn ML, Kirnbauer R, Roden RB, Lowy DR, Kast WM, Schiller JT (1998) Chimeric papillomavirus virus-like particles elicit antitumor immunity against the E7 oncoprotein in an HPV16 tumor model. Proc Natl Acad Sci USA 95:1800–1805

    Article  PubMed  CAS  Google Scholar 

  5. Jansen KU, Rosolowsky M, Schultz LD, Markus HZ, Cook JC, Donnelly JJ, Martinez D, Ellis RW, Shaw AR (1995) Vaccination with yeast-expressed cottontail rabbit papillomavirus (CRPV) virus-like particles protects rabbits from CRPV-induced papilloma formation. Vaccine 13:1509–1514

    Article  PubMed  CAS  Google Scholar 

  6. Kawana K, Yoshikawa H, Taketani Y, Yoshiike K, Kanda T (1998) In vitro construction of pseudovirions of human papillomavirus type 16: incorporation of plasmid DNA into reassembled L1/L2 capsids. J Virol 72:10298–10300

    PubMed  CAS  Google Scholar 

  7. Liu WJ, Liu XS, Zhao KN, Leggatt GR, Frazer IH (2000) Papillomavirus virus-like particles for the delivery of multiple cytotoxic T cell epitopes (in process citation). Virology 273:374–382

    Article  PubMed  CAS  Google Scholar 

  8. Liu XS, Liu WJ, Zhao KN, Liu YH, Leggatt G, Frazer IH (2002) Route of administration of chimeric BPV1 VLP determines the character of the induced immune responses. Immunol Cell Biol 80:21–29

    Article  PubMed  CAS  Google Scholar 

  9. Mach H, Volkin DB, Troutman RD, Wang B, Luo Z, Jansen KU, Shi L (2006) Disassembly and reassembly of yeast-derived recombinant human papillomavirus virus-like particles (HPV VLPs). J Pharm Sci 95:2195–2206

    Article  PubMed  CAS  Google Scholar 

  10. Muller M, Zhou J, Reed TD, Rittmuller C, Burger A, Gabelsberger J, Braspenning J, Gissmann L (1997) Chimeric papillomavirus-like particles. Virology 234:93–111

    Article  PubMed  CAS  Google Scholar 

  11. Nieland JD, Da Silva DM, Velders MP, de Visser KE, Schiller JT, Muller M, Kast WM (1999) Chimeric papillomavirus virus-like particles induce a murine self-antigen-specific protective and therapeutic antitumor immune response. J Cell Biochem 73:145–152

    Article  PubMed  CAS  Google Scholar 

  12. Peng S, Frazer IH, Fernando GJ, Zhou J (1998) Papillomavirus virus-like particles can deliver defined CTL epitopes to the MHC class I pathway. Virology 240:147–157

    Article  PubMed  CAS  Google Scholar 

  13. Roden RB, Greenstone HL, Kirnbauer R, Booy FP, Jessie J, Lowy DR, Schiller JT (1996) In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. J Virol 70:5875–5883

    PubMed  CAS  Google Scholar 

  14. Rossi JL, Gissmann L, Jansen K, Muller M (2000) Assembly of human papillomavirus type 16 pseudovirions in Saccharomyces cerevisiae. Hum Gene Ther 11:1165–1176

    Article  PubMed  CAS  Google Scholar 

  15. Schafer K, Muller M, Faath S, Henn A, Osen W, Zentgraf H, Benner A, Gissmann L, Jochmus I (1999) Immune response to human papillomavirus 16 L1E7 chimeric virus-like particles: induction of cytotoxic T cells and specific tumor protection. Int J Cancer 81:881–888

    Article  PubMed  CAS  Google Scholar 

  16. Suzich JA, Ghim SJ, Palmer-Hill FJ, White WI, Tamura JK, Bell JA, Newsome JA, Jenson AB, Schlegel R (1995) Systemic immunization with papillomavirus L1 protein completely prevents the development of viral mucosal papillomas. Proc Natl Acad Sci USA 92:11553–11557

    Article  PubMed  CAS  Google Scholar 

  17. Touze A, Coursaget P (1998) In vitro gene transfer using human papillomavirus-like particles. Nucleic Acids Res 26:1317–1323

    Article  PubMed  CAS  Google Scholar 

  18. Unckell F, Streeck RE, Sapp M (1997) Generation and neutralization of pseudovirions of human papillomavirus type 33. J Virol 71:2934–2939

    PubMed  CAS  Google Scholar 

  19. Varsani A, Williamson AL, de Villiers D, Becker I, Christensen ND, Rybicki EP (2003) Chimeric human papillomavirus type 16 (HPV-16) L1 particles presenting the common neutralizing epitope for the L2 minor capsid protein of HPV-6 and HPV-16. J Virol 77:8386–8393

    Article  PubMed  CAS  Google Scholar 

  20. Varsani A, Williamson AL, Jaffer MA, Rybicki EP (2006) A deletion and point mutation study of the human papillomavirus type 16 major capsid gene. Virus Res 122:154–163

    Article  PubMed  CAS  Google Scholar 

  21. Zhao KN, Sun XY, Frazer IH, Zhou J (1998) DNA packaging by L1 and L2 capsid proteins of bovine papillomavirus type 1. Virology 243:482–491

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We would like thank to Dr. Neil Christensen for providing the monoclonal antibodies used in this study. This research was funded by the Poliomyelitis Research Foundation of South Africa (grant #04/25) and the University of Cape Town start up grant. AV is supported by the Carnegie Corporation of New York.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, Cape Town, 7701, South Africa

    Oliver P. Windram, Edward P. Rybicki, Dionne N. Shepherd & Arvind Varsani

  2. Electron Microscope Unit, University of Cape Town, Rondebosch, Cape Town, 7701, South Africa

    Brandon Weber, Mohamed A. Jaffer & Arvind Varsani

Authors
  1. Oliver P. Windram
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  2. Brandon Weber
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  3. Mohamed A. Jaffer
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  4. Edward P. Rybicki
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  5. Dionne N. Shepherd
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  6. Arvind Varsani
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Corresponding author

Correspondence to Arvind Varsani.

Electronic supplementary material

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Supplementary Table 1: Primer sets used for cloning fusion constructs (DOC 37 kb)

705_2007_25_MOESM2_ESM.doc

Supplementary Figure 1: CsCl banding patterns observed for the fusion constructs. Two bands observed for GFP-L1 and L1-GFP constructs with the lower band fluorescing under UV (GFP). Three bands were observed for L1:GFP-L1 and L1:L2-GFP. (DOC 834 kb)

705_2007_25_MOESM3_ESM.doc

Supplementary Figure 2: Agarose gel stained with ethidium bromide, showing the banding of 6-8-kb DNA incorporated into the GFP fusion VLPs. A greater amount of DNA with a broader size range (6-8kb) was observed for GFP-L1 and L1-GFP. DNA of smaller size was observed in the L1:GFP-L2 and L1:L2-GFP VLPs. The fluorescence in the wells represents the GFP fusion VLPs. (DOC 114 kb)

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Windram, O.P., Weber, B., Jaffer, M.A. et al. An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins. Arch Virol 153, 585–589 (2008). https://doi.org/10.1007/s00705-007-0025-2

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  • DOI: https://doi.org/10.1007/s00705-007-0025-2

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