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Construction of HCV-polytope vaccine candidates harbouring immune-enhancer sequences and primary evaluation of their immunogenicity in BALB/c mice

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Abstract

An efficient vaccine against hepatitis-C virus (HCV) infection requires vigorous and focused CD8+ T-cell responses against viral antigens. Due to immunosuppressive effect of HCV antigens, polytope vaccines comprising the minimal CD8+CTL epitopes are of peculiar concern. Herein, to provide information for construction of efficient HCV polytope vaccine candidates, one H-2Dd (E2405–414:E2) and two HLA-A*0201 (E1363–372:E1 and Core35-44:C)-restricted CD8+ T-cell epitopes of HCV were selected. By employing number of in silico analyses, the E2E1C linear format was predicted as optimum epitope consecution and after amplification by SOEing-PCR, the corresponding DNA sequence was cloned in pcDNA3.1+ vector. To further evaluate the role of immune-enhancer elements, a universal T-helper epitope (PADRE), endoplasmic reticulum signal sequence (ERss) and hepatitis-B surface-antigen (HBsAg) gene were fused separately or in combination to the E2E1C minigene. In vitro analyses of polytopes by different DNA/protein-based assays demonstrated proper transcription/expression of constructs in transfected cells. Measurement of the HBsAg-mediated particle secretion by ELISA indicated lack of secretion in the related polytopes. Results of delayed-type hypersensitivity (DTH) as a preliminary in vivo analysis, and confirmatory ELISPOT assays showed the proper processing and presentation of H-2Dd-restricted-E2 epitope and approved the enhancing effect of PADRE and ERss sequences but not HBsAg for the immune responses against E2 in immunized BALB/c mice. Our results pointed to the value of in silico predictions and application of immune-enhancer elements as well as DTH analysis for design and primary in vivo evaluation of HCV polytopes, prior to costly transgenic studies on immunogenicity of HLA-A*0201 epitopes.

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References

  1. J.M. Pawlotsky, Nat. Rev. Gastroenterol. Hepatol. 6, 383–385 (2009)

    Article  CAS  Google Scholar 

  2. K. Lang, D.B. Weiner, Expert Rev. Vaccines 7, 915–923 (2008)

    Article  CAS  Google Scholar 

  3. S. Ishii, M.J. Koziel, Clin. Immunol. 128, 133–147 (2008)

    Article  CAS  Google Scholar 

  4. A.M. Arvin, H.B. Greenberg, Virology 344, 240–249 (2006)

    Article  CAS  Google Scholar 

  5. P. Martin, B. Simon, Y.C. Lone, L. Chatel, R. Barry, G. Inchauspe, A. Fournillier, Vaccine 26, 2471–2481 (2008)

    Article  CAS  Google Scholar 

  6. N. Himoudi, J.D. Abraham, A. Fournillier, Y.C. Lone, A. Joubert, A. Op De Beeck, D. Freida, F. Lemonnier, M.P. Kieny, G. Inchauspe, J. Virol. 76, 12735–12746 (2002)

    Article  CAS  Google Scholar 

  7. S.J. Ha, S.N. Mueller, E.J. Wherry, D.L. Barber, R.D. Aubert, A.H. Sharpe, G.J. Freeman, R. Ahmed, J. Exp. Med. 205, 543–555 (2008)

    Article  CAS  Google Scholar 

  8. A. Suhrbier, Expert Rev. Vaccines 1, 207–213 (2002)

    Article  CAS  Google Scholar 

  9. W.P. Woo, T. Doan, K.A. Herd, H.J. Netter, R.W. Tindle, J. Virol. 80, 3975–3984 (2006)

    Article  CAS  Google Scholar 

  10. J.N. Radcliffe, J.S. Roddick, P.S. Friedmann, F.K. Stevenson, S.M. Thirdborough, J. Immunol. 177, 6626–6633 (2006)

    Article  CAS  Google Scholar 

  11. T. Doan, K. Herd, I. Ramshaw, S. Thomson, R.W. Tindle, Cancer Immunol. Immunother. 54, 157–171 (2005)

    Article  CAS  Google Scholar 

  12. L. Shi, S. Liu, G.X. Fan, L. Sheng, H.X. Ren, Y.K. Yuan, Viral Immunol. 19, 702–711 (2006)

    Article  CAS  Google Scholar 

  13. S.A. Thomson, S.L. Elliott, M.A. Sherritt, K.W. Sproat, B.E. Coupar, A.A. Scalzo, C.A. Forbes, A.M. Ladhams, X.Y. Mo, R.A. Tripp, P.C. Doherty, D.J. Moss, A. Suhrbier, J. Immunol. 157, 822–826 (1996)

    CAS  PubMed  Google Scholar 

  14. Y. Luo, M.E. Dorf, in Current Protocols in Immunology, ed. by F.M. Ausubel, E. Brent, D.D. Moore, J.G. Seidman, J.A. Smith, K. Struhl (Wiley, New York, 1993), p. 4.5.1

    Google Scholar 

  15. A.K. Nussbaum, C. Kuttler, K.P. Hadeler, H.G. Rammensee, H. Schild, Immunogenetics 53, 87–94 (2001)

    Article  CAS  Google Scholar 

  16. C. Kesmir, A.K. Nussbaum, H. Schild, V. Detours, S. Brunak, Protein Eng. 15, 287–296 (2002)

    Article  CAS  Google Scholar 

  17. K.C. Parker, M.A. Bednarek, J.E. Coligan, J. Immunol. 152, 163–175 (1994)

    CAS  PubMed  Google Scholar 

  18. L. Deml, A. Bojak, S. Steck, M. Graf, J. Wild, R. Schirmbeck, H. Wolf, R. Wagner, J. Virol. 75, 10991–11001 (2001)

    Article  CAS  Google Scholar 

  19. J. Carter, Production of Anti-Peptide Antisera (Wiley, New York, 2003)

    Book  Google Scholar 

  20. H. Towbin, T. Staehelin, J. Gordon, Proc. Natl Acad. Sci. USA 76, 4350–4354 (1979)

    Article  CAS  Google Scholar 

  21. S. Radhakrishnan, P. Abraham, G. Sridharan, Diagn. Microbiol. Infect. Dis. 33, 153–156 (1999)

    Article  CAS  Google Scholar 

  22. A. Baez-Astua, E. Herraez-Hernandez, N. Garbi, H.A. Pasolli, V. Juarez, H. Zur Hausen, A. Cid-Arregui, J. Virol. 79, 12807–12817 (2005)

    Article  CAS  Google Scholar 

  23. J. Sambrook, D.W. Russell, in Molecular Cloning, a Laboratory Manual (CSHL Press, New York, 2001), pp. 17.48–17.51

  24. F. Roohvand, M.R. Aghasadeghi, S.M. Sadat, A. Budkowska, A.R. Khabiri, Biochem. Biophys. Res. Commun. 354, 641–649 (2007)

    Article  CAS  Google Scholar 

  25. Y. Chen, R.G. Webster, D.L. Woodland, J. Immunol. 160, 2425–2432 (1998)

    CAS  PubMed  Google Scholar 

  26. M. Del Val, H.J. Schlicht, T. Ruppert, M.J. Reddehase, U.H. Koszinowski, Cell 66, 1145–1153 (1991)

    Article  CAS  Google Scholar 

  27. A. Craiu, T. Akopian, A. Goldberg, K.L. Rock, Proc. Natl Acad. Sci. USA 94, 10850–10855 (1997)

    Article  CAS  Google Scholar 

  28. A.L. Puaux, D. Marsac, S. Prost, M.K. Singh, P. Earl, B. Moss, R. Le Grand, Y. Riviere, M.L. Michel, Vaccine 22, 3535–3545 (2004)

    Article  CAS  Google Scholar 

  29. J. Jin, J.Y. Yang, J. Liu, Y.Y. Kong, Y. Wang, G.D. Li, World J. Gastroenterol. 8, 505–510 (2002)

    Article  CAS  Google Scholar 

  30. J. Hui, M. Mancini, G. Li, Y. Wang, P. Tiollais, M.L. Michel, Vaccine 17, 1711–1718 (1999)

    Article  CAS  Google Scholar 

  31. H. Bisht, D.A. Chugh, S. Swaminathan, N. Khanna, Protein Expr. Purif. 23, 84–96 (2001)

    Article  CAS  Google Scholar 

  32. M. Michel, Y.C. Lone, M. Centlivre, P. Roux, S. Wain-Hobson, M. Sala, Vaccine 25, 1901–1911 (2007)

    Article  CAS  Google Scholar 

  33. S. Phogat, K. Svehla, M. Tang, A. Spadaccini, J. Muller, J. Mascola, I. Berkower, R. Wyatt, Virology 373, 72–84 (2008)

    Article  CAS  Google Scholar 

  34. T.L. Whiteside, Y. Zhao, T. Tsukishiro, E.M. Elder, W. Gooding, J. Baar, Clin. Cancer Res. 9, 641–649 (2003)

    CAS  PubMed  Google Scholar 

  35. M.P. Velders, S. Weijzen, G.L. Eiben, A.G. Elmishad, P.M. Kloetzel, T. Higgins, R.B. Ciccarelli, M. Evans, S. Man, L. Smith, W.M. Kast, J. Immunol. 166, 5366–5373 (2001)

    Article  CAS  Google Scholar 

  36. W. Xu, Y. Chu, R. Zhang, H. Xu, Y. Wang, S. Xiong, Virology 334, 255–263 (2005)

    Article  CAS  Google Scholar 

  37. A. Sette, J. Fikes, Curr. Opin. Immunol. 15, 461–470 (2003)

    Article  CAS  Google Scholar 

  38. D. Marsac, A.L. Puaux, Y. Riviere, M.L. Michel, Immunobiology 210, 305–319 (2005)

    Article  CAS  Google Scholar 

  39. R.E. Toes, E.I. van der Voort, S.P. Schoenberger, J.W. Drijfhout, L. van Bloois, G. Storm, W.M. Kast, R. Offringa, C.J. Melief, J. Immunol. 160, 4449–4456 (1998)

    CAS  PubMed  Google Scholar 

  40. A. Lanzavecchia, F. Sallusto, Nat. Immunol. 2, 487–492 (2001)

    Article  CAS  Google Scholar 

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Acknowledgements

A.A. received a fellowship from graduate school of Institute Pasteur to pursue this study in the Ph.D. programme. Portions of this study were presented as oral presentations in symposium of Research on Infectious Diseases—A Global Challenge (26–27 June 2008, Paris) and the first international vaccine congress (9–11 December 2007, Amsterdam). This work was financially supported by Pasteur Institute of Iran.

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Authors and Affiliations

  1. Hepatitis & AIDS Department-NRGB Lab, Pasteur Institute of Iran, Pasteur Ave, Tehran, 1316943551, Iran

    Arash Arashkia, Farzin Roohvand, Arash Memarnejadian & Mohammad Reza Aghasadeghi

  2. Molecular Immunology and Vaccine Research Lab, Pasteur Institute of Iran, Tehran, Iran

    Sima Rafati

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  1. Arash Arashkia
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  2. Farzin Roohvand
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  3. Arash Memarnejadian
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  4. Mohammad Reza Aghasadeghi
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Correspondence to Farzin Roohvand.

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Arashkia, A., Roohvand, F., Memarnejadian, A. et al. Construction of HCV-polytope vaccine candidates harbouring immune-enhancer sequences and primary evaluation of their immunogenicity in BALB/c mice. Virus Genes 40, 44–52 (2010). https://doi.org/10.1007/s11262-009-0417-3

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  • DOI: https://doi.org/10.1007/s11262-009-0417-3

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