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Identification of immunogenic regions within the alternative reading frame protein of hepatitis C virus (genotype 3)

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Abstract

Hepatitis C virus (HCV) encodes ten classic proteins as well as a newly discovered alternative reading frame protein (ARFP) whose synthesis originates from the core region by a +1 frameshift. ARFP is produced by all HCV genotypes, but its function remains unknown. Although the immunogenicity of genotype 1- and 2-derived ARFP in infected hosts has been reported, no information is available for genotype 3-encoded ARFP. HCV genotype 3 core/ARFP region was PCR amplified, cloned, and sequenced. Recombinant ARFP and peptides were employed in ELISAs with patient serum samples. The effect of peptides on peripheral blood mononucleocytes (PBMCs) was also studied. DNA cloning and sequencing of HCV genotype 3 strain (PKHCV3) revealed it to encode 160 aa ARFP, which harbors a C-terminal extension of 36 aa. Serum from 74 of 88 patients (84%) contained rARFP-reactive antibodies. Peptide ELISAs showed that all regions of rARFP were immunogenic, with peptide F7 (DSLSPRRAGAKAGPGLSPGT) being the most immunodominant. When incubated with PBMCs from HCV-infected individuals, F7 stimulated the production of TNFα and IL10. PKHCV3-derived ARFP encodes a 160 aa protein and antibodies against its entire length are found in 84% of all genotype 3-infected subjects. Peptide ELISAs revealed F7 to be highly immunogenic and capable of eliciting impressive T-cell responses.

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References

  1. Qureshi SA, Qureshi H, Hameed A (2009) Hepatitis C therapy—the future looks bright. Eur J Clin Microbiol Infect Dis 28(12):1409–1413

    Article  PubMed  CAS  Google Scholar 

  2. Qureshi SA (2007) Hepatitis C virus—biology, host evasion strategies, and promising new therapies on the horizon. Med Res Rev 27(3):353–373

    Article  PubMed  CAS  Google Scholar 

  3. Branch AD, Stump DD, Gutierrez JA, Eng F, Walewski JL (2005) The hepatitis C virus alternate reading frame (ARF) and its family of novel products: the alternate reading frame protein/F-protein, the double-frameshift protein, and others. Semin Liver Dis 25(1):105–117

    Article  PubMed  CAS  Google Scholar 

  4. Vassilaki N, Mavromara P (2009) The HCV ARFP/F/core+1 protein: production and functional analysis of an unconventional viral product. IUBMB Life 61(7):739–752

    Article  PubMed  CAS  Google Scholar 

  5. Vassilaki N, Mavromara P (2003) Two alternative translation mechanisms are responsible for the expression of the HCV ARFP/F/core+1 coding open reading frame. J Biol Chem 278(42):40503–40513

    Article  PubMed  CAS  Google Scholar 

  6. Baril M, Brakier-Gingras L (2005) Translation of the F protein of hepatitis C virus is initiated at a non-AUG codon in a +1 reading frame relative to the polyprotein. Nucleic Acids Res 33(5):1474–1486

    Article  PubMed  CAS  Google Scholar 

  7. Boulant S, Becchi M, Penin F, Lavergne JP (2003) Unusual multiple recoding events leading to alternative forms of hepatitis C virus core protein from genotype 1b. J Biol Chem 278(46):45785–45792

    Article  PubMed  CAS  Google Scholar 

  8. Ratinier M, Boulant S, Combet C, Targett-Adams P, McLauchlan J, Lavergne JP (2008) Transcriptional slippage prompts recoding in alternate reading frames in the hepatitis C virus (HCV) core sequence from strain HCV-1. J Gen Virol 89(Pt 7):1569–1578

    Article  PubMed  CAS  Google Scholar 

  9. Yuksek K, Chen WL, Chien D, Ou JH (2009) Ubiquitin-independent degradation of hepatitis C virus F protein. J Virol 83(2):612–621

    Article  PubMed  CAS  Google Scholar 

  10. Boumlic A, Nominé Y, Charbonnier S, Dalagiorgou G, Vassilaki N, Kieffer B, Travé G, Mavromara P, Orfanoudakis G (2010) Prevalence of intrinsic disorder in the hepatitis C virus ARFP/Core+1/S protein. FEBS J 277(3):774–789

    Article  PubMed  CAS  Google Scholar 

  11. Vassilaki N, Friebe P, Meuleman P, Kallis S, Kaul A, Paranhos-Baccalà G, Leroux-Roels G, Mavromara P, Bartenschlager R (2008) Role of the hepatitis C virus core+1 open reading frame and core cis-acting RNA elements in viral RNA translation and replication. J Virol 82(23):11503–11515

    Article  PubMed  CAS  Google Scholar 

  12. Ma HC, Lin TW, Li H, Iguchi-Ariga SM, Ariga H, Chuang YL, Ou JH, Lo SY (2008) Hepatitis C virus ARFP/F protein interacts with cellular MM-1 protein and enhances the gene trans-activation activity of c-Myc. J Biomed Sci 15(4):417–425

    Article  PubMed  CAS  Google Scholar 

  13. Basu A, Steele R, Ray R, Ray RB (2004) Functional properties of a 16 kDa protein translated from an alternative open reading frame of the core-encoding genomic region of hepatitis C virus. J Gen Virol 85(Pt 8):2299–2306

    Article  PubMed  CAS  Google Scholar 

  14. Dou J, Liu P, Wang J, Zhang X (2006) Effect of hepatitis C virus core shadow protein expressed in human hepatoma cell line on human gene expression profiles. J Gastroenterol Hepatol 21(12):1794–1800

    Article  PubMed  CAS  Google Scholar 

  15. Huang YP, Cheng J, Zhang SL, Wang L, Guo J, Liu Y, Yang Y, Zhang LY, Bai GQ, Gao XS, Ji D, Lin SM, Shao Q (2005) Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system. World J Gastroenterol 11(36):5659–5665

    PubMed  CAS  Google Scholar 

  16. Wolf M, Dimitrova M, Baumert TF, Schuster C (2008) The major form of hepatitis C virus alternate reading frame protein is suppressed by core protein expression. Nucleic Acids Res 36(9):3054–3064

    Article  PubMed  CAS  Google Scholar 

  17. Shao SW, Wu WB, Bian ZQ, Yu JG, Zhao P, Zhao LJ, Zhu SY, Qi ZT (2009) Hepatitis C virus F protein inhibits cell apoptosis by activation of intracellular NF-kappaB pathway. Hepatol Res 39(3):282–289

    Article  PubMed  CAS  Google Scholar 

  18. Bain C, Parroche P, Lavergne JP, Duverger B, Vieux C, Dubois V, Komurian-Pradel F, Trépo C, Gebuhrer L, Paranhos-Baccalà G, Penin F, Inchauspé G (2004) Memory T-cell-mediated immune responses specific to an alternative core protein in hepatitis C virus infection. J Virol 78(19):10460–10469

    Article  PubMed  CAS  Google Scholar 

  19. Gao DY, Zhang XX, Hou G, Jin GD, Deng Q, Kong XF, Zhang DH, Ling Y, Yu DM, Gong QM, Zhan Q, Yao BL, Lu ZM (2008) Assessment of specific antibodies to F protein in serum samples from Chinese hepatitis C patients treated with interferon plus ribavarin. J Clin Microbiol 46(11):3746–3751

    Article  PubMed  CAS  Google Scholar 

  20. Chuang WC, Allain JP (2008) Differential reactivity of putative genotype 2 hepatitis C virus F protein between chronic and recovered infections. J Gen Virol 89(Pt 8):1890–1900

    Article  PubMed  CAS  Google Scholar 

  21. Xu Z, Choi J, Yen TS, Lu W, Strohecker A, Govindarajan S, Chien D, Selby MJ, Ou J (2001) Synthesis of a novel hepatitis C virus protein by ribosomal frameshift. EMBO J 20(14):3840–3848

    Article  PubMed  CAS  Google Scholar 

  22. Varaklioti A, Vassilaki N, Georgopoulou U, Mavromara P (2002) Alternate translation occurs within the core coding region of the hepatitis C viral genome. J Biol Chem 277(20):17713–17721

    Article  PubMed  CAS  Google Scholar 

  23. Walewski JL, Keller TR, Stump DD, Branch AD (2001) Evidence for a new hepatitis C virus antigen encoded in an overlapping reading frame. RNA 7(5):710–721

    Article  PubMed  CAS  Google Scholar 

  24. Cohen M, Bachmatov L, Ben-Ari Z, Rotman Y, Tur-Kaspa R, Zemel R (2007) Development of specific antibodies to an ARF protein in treated patients with chronic HCV infection. Dig Dis Sci 52(9):2427–2432

    Article  PubMed  CAS  Google Scholar 

  25. Komurian-Pradel F, Rajoharison A, Berland JL, Khouri V, Perret M, Van Roosmalen M, Pol S, Negro F, Paranhos-Baccalà G (2004) Antigenic relevance of F protein in chronic hepatitis C virus infection. Hepatology 40(4):900–909

    PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Najeeha Talat (Department of Pathology & Microbiology) and Mr. Junaid Iqbal and Ms. Hina Zuberi (Department of Biological & Biomedical Sciences) for their technical assistance. This work was funded by grants from the Higher Education Commission (HEC-20-342) and Aga Khan University Research Council (1UM/70139).

Conflict of interest

All authors declare that they have no conflict of interest.

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Author notes

  1. S. A. Qureshi

    Present address: Department of Biology, School of Science & Engineering, Lahore University of Management Sciences, DHA Lahore Cantt, 54792, Lahore, Pakistan

Authors and Affiliations

  1. Department of Biological and Biomedical Sciences, Aga Khan University, Stadium Road, Karachi, 74800, Pakistan

    H. Qureshi & S. A. Qureshi

  2. Department of Pathology and Microbiology, Aga Khan University, Stadium Road, Karachi, 74800, Pakistan

    R. Qazi

  3. Department of Medicine, Aga Khan University, Stadium Road, Karachi, 74800, Pakistan

    S. Hamid

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  1. H. Qureshi
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  2. R. Qazi
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  4. S. A. Qureshi
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Correspondence to S. A. Qureshi.

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Qureshi, H., Qazi, R., Hamid, S. et al. Identification of immunogenic regions within the alternative reading frame protein of hepatitis C virus (genotype 3). Eur J Clin Microbiol Infect Dis 30, 1075–1083 (2011). https://doi.org/10.1007/s10096-011-1194-1

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  • DOI: https://doi.org/10.1007/s10096-011-1194-1

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