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Archives of Virology

, Volume 160, Issue 1, pp 141–152 | Cite as

Enhancement of HCV polytope DNA vaccine efficacy by fusion to an N-terminal fragment of heat shock protein gp96

  • Leila Pishraft-Sabet
  • Anna D. Kosinska
  • Sima Rafati
  • Azam Bolhassani
  • Tahereh Taheri
  • Arash Memarnejadian
  • Seyed-Moayed Alavian
  • Michael Roggendorf
  • Katayoun Samimi-RadEmail author
Original Article

Abstract

Induction of a strong hepatitis C virus (HCV)-specific immune response plays a key role in control and clearance of the virus. A polytope (PT) DNA vaccine containing B- and T-cell epitopes could be a promising vaccination strategy against HCV, but its efficacy needs to be improved. The N-terminal domain of heat shock protein gp96 (NT(gp96)) has been shown to be a potent adjuvant for enhancing immunity. We constructed a PT DNA vaccine encoding four HCV immunodominant cytotoxic T lymphocyte epitopes (two HLA-A2- and two H2-Dd-specific motifs) from the Core, E2, NS3 and NS5B antigens in addition to a T-helper CD4+ epitope from NS3 and a B-cell epitope from E2. The NT(gp96) was fused to the C- or N-terminal end of the PT DNA (PT-NT(gp96) or NT(gp96)-PT), and their potency was compared. Cellular and humoral immune responses against the expressed peptides were evaluated in CB6F1 mice. Our results showed that immunization of mice with PT DNA vaccine fused to NT(gp96) induced significantly stronger T-cell and antibody responses than PT DNA alone. Furthermore, the adjuvant activity of NT(gp96) was more efficient in the induction of immune responses when fused to the C-terminal end of the HCV DNA polytope. In conclusion, the NT(gp96) improved the efficacy of the DNA vaccine, and this immunomodulatory effect was dependent on the position of the fusion.

Keywords

Enhanced Green Fluorescent Protein Humoral Immune Response Heat Shock Protein Gp96 Enhanced Green Fluorescent Protein Signal CB6F1 Mouse 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This study was funded and supported by Grant No. 13270 from Tehran University of Medical Sciences (TUMS) and Grant No. 90-803 from Baqiyatallah University of Medical Science.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Roohvand F, Kossari N (2011) Advances in hepatitis C virus vaccines, part one: advances in basic knowledge for hepatitis C virus vaccine design. Expert Opin Ther Pat 21:1811–1830PubMedCrossRefGoogle Scholar
  2. 2.
    Roohvand F, Kossari N (2012) Advances in hepatitis C virus vaccines, part two: advances in hepatitis C virus vaccine formulations and modalities. Expert Opin Ther Pat 22:391–415PubMedCrossRefGoogle Scholar
  3. 3.
    Cerny A, Chisari FV (1999) Pathogenesis of chronic hepatitis C: immunological features of hepatic injury and viral persistence. Hepatology 30:595–601PubMedCrossRefGoogle Scholar
  4. 4.
    Pestka JM, Zeisel MB, Blaser E, Schurmann P, Bartosch B, Cosset FL, Patel AH, Meisel H, Baumert J, Viazov S, Rispeter K, Blum HE, Roggendorf M, Baumert TF (2007) Rapid induction of virus-neutralizing antibodies and viral clearance in a single-source outbreak of hepatitis C. Proc Natl Acad Sci USA 104:6025–6030PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Suhrbier A (2002) Polytope vaccines for the codelivery of multiple CD8 T-cell epitopes. Expert Rev Vaccines 1:207–213PubMedCrossRefGoogle Scholar
  6. 6.
    Memarnejadian A, Roohvand F (2010) Fusion of HBsAg and prime/boosting augment Th1 and CTL responses to HCV polytope DNA vaccine. Cell Immunol 261:93–98PubMedCrossRefGoogle Scholar
  7. 7.
    Martin P, Simon B, Lone YC, Chatel L, Barry R, Inchauspe G, Fournillier A (2008) A vector-based minigene vaccine approach results in strong induction of T-cell responses specific of hepatitis C virus. Vaccine 26:2471–2481PubMedCrossRefGoogle Scholar
  8. 8.
    Doody AD, Kovalchin JT, Mihalyo MA, Hagymasi AT, Drake CG, Adler AJ (2004) Glycoprotein 96 can chaperone both MHC class I- and class II-restricted epitopes for in vivo presentation, but selectively primes CD8+ T cell effector function. J Immunol 172:6087–6092PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Javid B, MacAry PA, Lehner PJ (2007) Structure and function: heat shock proteins and adaptive immunity. J Immunol 179:2035–2040PubMedCrossRefGoogle Scholar
  10. 10.
    Yang Y, Liu B, Dai J, Srivastava PK, Zammit DJ, Lefrancois L, Li Z (2007) Heat shock protein gp96 is a master chaperone for toll-like receptors and is important in the innate function of macrophages. Immunity 26:215–226PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Baker-LePain JC, Sarzotti M, Fields TA, Li CY, Nicchitta CV (2002) GRP94 (gp96) and GRP94N-terminal geldanamycin binding domain elicit tissue nonrestricted tumor suppression. J Exp Med 196:1447–1459PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Li H, Zhou M, Han J, Zhu X, Dong T, Gao GF, Tien P (2005) Generation of murine CTL by a hepatitis B virus-specific peptide and evaluation of the adjuvant effect of heat shock protein glycoprotein 96 and its terminal fragments. J Immunol 174:195–204PubMedCrossRefGoogle Scholar
  13. 13.
    Lohr HF, Schmitz D, Arenz M, Weyer S, Gerken G, Meyer zum Buschenfelde KH (1999) The viral clearance in interferon-treated chronic hepatitis C is associated with increased cytotoxic T cell frequencies. World J Hepatol 31:407–415CrossRefGoogle Scholar
  14. 14.
    Shirai M, Okada H, Nishioka M, Akatsuka T, Wychowski C, Houghten R, Pendleton CD, Feinstone SM, Berzofsky JA (1994) An epitope in hepatitis C virus core region recognized by cytotoxic T cells in mice and humans. J Virol 68:3334–3342PubMedCentralPubMedGoogle Scholar
  15. 15.
    Park SH, Yang SH, Lee CG, Youn JW, Chang J, Sung YC (2003) Efficient induction of T helper 1 CD4+ T-cell responses to hepatitis C virus core and E2 by a DNA prime-adenovirus boost. Vaccine 21:4555–4564PubMedCrossRefGoogle Scholar
  16. 16.
    Chang KM, Thimme R, Melpolder JJ, Oldach D, Pemberton J, Moorhead-Loudis J, McHutchison JG, Alter HJ, Chisari FV (2001) Differential CD4(+) and CD8(+) T-cell responsiveness in hepatitis C virus infection. Hepatology 33:267–276PubMedCrossRefGoogle Scholar
  17. 17.
    Zhu F, Eckels DD (2002) Functionally distinct helper T-cell epitopes of HCV and their role in modulation of NS3-specific, CD8+/tetramer positive CTL. Hum Immunol 63:710–718PubMedCrossRefGoogle Scholar
  18. 18.
    Zhang P, Zhong L, Struble EB, Watanabe H, Kachko A, Mihalik K, Virata-Theimer ML, Alter HJ, Feinstone S, Major M (2009) Depletion of interfering antibodies in chronic hepatitis C patients and vaccinated chimpanzees reveals broad cross-genotype neutralizing activity. Proc Natl Acad Sci USA 106:7537–7541PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Bolhassani A, Zahedifard F, Taslimi Y, Taghikhani M, Nahavandian B, Rafati S (2009) Antibody detection against HPV16 E7 & GP96 fragments as biomarkers in cervical cancer patients. Indian J Med Res 130:533–541PubMedGoogle Scholar
  20. 20.
    Tavakoli-Yaraki M, Karami-Tehrani F, Salimi V, Sirati-Sabet M (2013) Induction of apoptosis by trichostatin A in human breast cancer cell lines: involvement of 15-Lox-1. Tumour Biol 34:241–249PubMedCrossRefGoogle Scholar
  21. 21.
    Lu M, Isogawa M, Xu Y, Hilken G (2005) Immunization with the gene expressing woodchuck hepatitis virus nucleocapsid protein fused to cytotoxic-T-lymphocyte-associated antigen 4 leads to enhanced specific immune responses in mice and woodchucks. J Virol 79:6368–6376PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Kosinska AD, Zhang E, Johrden L, Liu J, Seiz PL, Zhang X, Ma Z, Kemper T, Fiedler M, Glebe D, Wildner O, Dittmer U, Lu M, Roggendorf M (2013) Combination of DNA prime adenovirus boost immunization with entecavir elicits sustained control of chronic hepatitis B in the woodchuck model. PLoS Pathog 9:e100339. doi: 10.1371/journal.ppat.1003391 CrossRefGoogle Scholar
  23. 23.
    Zelinskyy G, Dietze K, Sparwasser T, Dittmer U (2009) Regulatory T cells suppress antiviral immune responses and increase viral loads during acute infection with a lymphotropic retrovirus. PLoS Pathog 5:e1000406. doi: 10.1371/journal.ppat.1000406 PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Boettler T, Spangenberg HC, Neumann-Haefelin C, Panther E, Urbani S, Ferrari C, Blum HE, von Weizsacker F, Thimme R (2005) T cells with a CD4+ CD25+ regulatory phenotype suppress in vitro proliferation of virus-specific CD8+ T cells during chronic hepatitis C virus infection. J Virol 79:7860–7867PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Rushbrook SM, Ward SM, Unitt E, Vowler SL, Lucas M, Klenerman P, Alexander GJ (2005) Regulatory T cells suppress in vitro proliferation of virus-specific CD8+ T cells during persistent hepatitis C virus infection. J Virol 79:7852–7859PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Bolhassani A, Zahedifard F, Taghikhani M, Rafati S (2008) Enhanced immunogenicity of HPV16 E7 accompanied by Gp96 as an adjuvant in two vaccination strategies. Vaccine 26:3362–3370PubMedCrossRefGoogle Scholar
  27. 27.
    Daemi A, Bolhassani A, Rafati S, Zahedifard F, Hosseinzadeh S, Doustdari F (2012) Different domains of glycoprotein 96 influence HPV16 E7 DNA vaccine potency via electroporation mediated delivery in tumor mice model. Immunol Lett 148:117–125PubMedCrossRefGoogle Scholar
  28. 28.
    Li HT, Yan JB, Li J, Zhou MH, Zhu XD, Zhang YX, Tien P (2005) Enhancement of humoral immune responses to HBsAg by heat shock protein gp96 and its N-terminal fragment in mice. World J Gastroenterol 11:2858–2863PubMedGoogle Scholar
  29. 29.
    Basu S, Binder RJ, Ramalingam T, Srivastava PK (2001) CD91 is a common receptor for heat shock proteins gp96, hsp90, hsp70, and calreticulin. Immunity 14:303–313PubMedCrossRefGoogle Scholar
  30. 30.
    Yan J, Liu X, Wang Y, Jiang X, Liu H, Wang M, Zhu X, Wu M, Tien P (2007) Enhancing the potency of HBV DNA vaccines using fusion genes of HBV-specific antigens and the N-terminal fragment of gp96. J Gene Med 9:107–121PubMedCrossRefGoogle Scholar
  31. 31.
    Mohit E, Bolhassani A, Zahedifard F, Taslimi Y, Rafati S (2011) The contribution of NT-gp96 as an adjuvant for increasing HPV16 E7-specific immunity in C57BL/6 mouse model. Scand J Immunol 75:27–37CrossRefGoogle Scholar
  32. 32.
    Wang S, Qiu L, Liu G, Li Y, Zhang X, Jin W, Gao GF, Kong X, Meng S (2011) Heat shock protein gp96 enhances humoral and T cell responses, decreases Treg frequency and potentiates the anti-HBV activity in BALB/c and transgenic mice. Vaccine 29:6342–6351PubMedCrossRefGoogle Scholar
  33. 33.
    Cooper S, Erickson AL, Adams EJ, Kansopon J, Weiner AJ, Chien DY, Houghton M, Parham P, Walker CM (1999) Analysis of a successful immune response against hepatitis C virus. Immunity 10:439–449PubMedCrossRefGoogle Scholar
  34. 34.
    Seder RA, Darrah PA, Roederer M (2008) T-cell quality in memory and protection: implications for vaccine design. Nat Rev Immunol 8:247–258PubMedCrossRefGoogle Scholar
  35. 35.
    Ahmed R, Akondy RS (2011) Insights into human CD8(+) T-cell memory using the yellow fever and smallpox vaccines. Immunol Cell Biol 89:340–345PubMedCrossRefGoogle Scholar
  36. 36.
    Li S, Roberts S, Plebanski M, Gouillou M, Spelman T, Latour P, Jackson D, Brown L, Sparrow RL, Prince HM, Hart D, Loveland BE, Gowans EJ (2012) Induction of multi-functional T cells in a phase I clinical trial of dendritic cell immunotherapy in hepatitis C virus infected individuals. PLoS One 7:e39368. doi: 10.1371/journal.pone.0039368 PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Houghton M, Abrignani S (2005) Prospects for a vaccine against the hepatitis C virus. Nature 436:961–966PubMedCrossRefGoogle Scholar
  38. 38.
    Chen C, Li J, Bi Y, Yang L, Meng S, Zhou Y, Jia X, Sun L, Liu W (2013) Synthetic B- and T-cell epitope peptides of porcine reproductive and respiratory syndrome virus with Gp96 as adjuvant induced humoral and cell-mediated immunity. Vaccine 31:1838–1847PubMedCrossRefGoogle Scholar
  39. 39.
    Pakravan N, Hashemi SM, Hassan ZM (2011) Adjuvant activity of GP96 C-terminal domain towards Her2/neu DNA vaccine is fusion direction-dependent. Cell Stress Chaperones 16:41–48PubMedCentralPubMedCrossRefGoogle Scholar
  40. 40.
    Hoeller D, Dikic I (2009) Targeting the ubiquitin system in cancer therapy. Nature 458:438–444PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Leila Pishraft-Sabet
    • 1
    • 2
  • Anna D. Kosinska
    • 3
  • Sima Rafati
    • 4
  • Azam Bolhassani
    • 5
  • Tahereh Taheri
    • 4
  • Arash Memarnejadian
    • 5
  • Seyed-Moayed Alavian
    • 6
  • Michael Roggendorf
    • 3
  • Katayoun Samimi-Rad
    • 1
    Email author
  1. 1.Department of Virology, School of Public HealthTehran University of Medical SciencesTehranIslamic Republic of Iran
  2. 2.Razi Vaccine and Serum Research InstituteKarajIran
  3. 3.Institute of VirologyUniversity Hospital of EssenEssenGermany
  4. 4.Molecular Immunology and Vaccine Research LaboratoryPasteur Institute of IranTehranIran
  5. 5.Hepatitis and HIV LaboratoryPasteur Institute of IranTehranIran
  6. 6.Research Center for Gastroenterology and Liver DiseaseBaqiyatallah University of Medical SciencesTehranIran

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