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Molecular Biology

, Volume 40, Issue 2, pp 312–323 | Cite as

Mapping of antigenic determinants of hepatitis C virus proteins using phage display

  • E. A. Rechkina
  • G. F. Denisova
  • O. V. Masalova
  • L. F. Lideman
  • D. A. Denisov
  • E. I. Lesnova
  • R. I. Ataullakhanov
  • S. V. Gurianova
  • A. A. Kushch
Miscellaneous

Abstract

Analysis of the properties for individual hepatitis C virus (HCV) proteins makes it possible to establish their molecular structure and conformation, to localize antigenic and immunogenic determinants, to identify protective epitopes, and to solve applied problems (e.g., design of diagnostic tests, vaccines, and drugs). Linear and conformational epitopes of HCV proteins were localized using the phage display technique, and the peptides exposed on the phages selected with monoclonal antibodies against HCV proteins were tested for immunogenicity. Of the 11 epitopes revealed, three were strongly linear; two depended on the secondary; and one on the tertiary structure of the corresponding protein (conformational epitopes). Amino acid sequences involved in the other epitopes were established. The results can be used to improve the diagnosis of hepatitis C, to study the effect of amino acid substitutions on the antigenic properties of HCV proteins, and to analyze the immune response in patients infected with genotypically different HCV. It was shown with the example of the NS5A epitope that phage particles with epitope-mimicking peptides (mimotopes) induce production of antibodies against the corresponding HCV proteins.

Key words

hepatitis C virus proteins antigenic determinants epitope mapping phage display immunogenic properties 

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References

  1. 1.
    Laperche S., Courouce A.M. 1997. Development of a biological diagnosis for hepatitis C. Transfus. Clin. Biol. 4, 291–298.CrossRefPubMedGoogle Scholar
  2. 2.
    Bartenschlager R., Lohmann V. 2000. Replication of hepatitis C virus. J. Gen. Virol. 81, 1631–1648.PubMedGoogle Scholar
  3. 3.
    Moradpour D., Kary P., Rice C.M., Blum H.E. 1998. Continuous human cell lines inducibly expressing hepatitis C virus structural and nonstructural proteins. Hepatology. 28, 193–201.CrossRefGoogle Scholar
  4. 4.
    Chang J.C., Ruedinger B., Cong M., Lambert S., Lopareva E., Purdy M., Holloway B.P., Jue D.L., Ofenloch B., Fields H.A., Khudyakov Y.E. 1999. Artificial NS4 mosaic antigen of hepatitis C virus. J. Med. Virol. 59, 437–450.CrossRefPubMedGoogle Scholar
  5. 5.
    Rosa C., Osborne S., Garetto F., Griva S., Rivella A., Calabresi G., Guaschino R., Bonelli F. 1995. Epitope mapping of the NS4 and NS5 gene products of hepatitis C virus and the use of a chimeric NS4-NS5 synthetic peptide for serodiagnosis. J. ViroI. Meth. 55, 219–232.Google Scholar
  6. 6.
    Courouce A.M., Noel L., Barin F., Elghouzzi M.H., Lunel F., North M.L., Smilovici W. 1998. A comparative evaluation of the sensitivity of five anti-hepatitis C virus immunoblot assays. Vox Sang. 74, 217–224.CrossRefPubMedGoogle Scholar
  7. 7.
    Schwab C., Twardek A., Lo T.P., Brayer D., Bosshard H.R. 1993. Mapping antibody binding sites on cytochrome c with synthetic peptides: Are results representative of the antigenic structure of proteins? Protein Sci. 2, 175–182.PubMedGoogle Scholar
  8. 8.
    Pereboeva L.A., Pereboev A.V., Morris G.E. 1998. Identification of antigenic sites on three hepatitis C virus proteins using phage-displayed peptide libraries. J. Med. Virol. 56, 105–111.CrossRefPubMedGoogle Scholar
  9. 9.
    Khudyakov Yu.E., Khudyakova N.S., Jue D.L., Lambert S.B., Fang S., Fields H.A. 1995. Linear B-cell epitopes of the NS3-NS4-NS5 proteins of the hepatitis C virus as modeled with synthetic peptides. Virology. 206, 666–672.CrossRefGoogle Scholar
  10. 10.
    Jolivet-Reynaud C., Dalbon P., Viola F., Yvon S., Paranhos-Baccala G., Piga N., Bridon L., Trabaud M.A., Battail N., Sibai G., Jolivet M. 1998. HCV core immunodominant region analysis using mouse monoclonal antibodies and human sera: Characterization of major epitopes useful for antigen detection. J. Med. Virol. 56, 300–309.CrossRefPubMedGoogle Scholar
  11. 11.
    Scott J.K., Smith G.P. 1990. Searching for peptide ligands with an epitope library. Science. 249, 386–390.PubMedGoogle Scholar
  12. 12.
    Jolivet-Reynaud C., Adida A., Michel S., Deléage G., Paranhos-Baccala G., Gonin V., Battail-Poirot N., Lacoux X., Rolland D. 2004. Characterization of mimotopes mimicking an immunodominant conformational epitope on the hepatitis C virus NS3 helicase. J. Med. Virol. 72, 385–395.CrossRefPubMedGoogle Scholar
  13. 13.
    Prezzi C., Nuzzo M., Meola A., Delmastro P., Galfre G., Cortese R., Nicosia A., Monaci P. 1996. Selection of antigenic and immunogenic mimics of hepatitis C virus using sera from patients. J. Immunol. 156, 4504–4513.PubMedGoogle Scholar
  14. 14.
    Masalova O.V., Lakina E.I., Abdulmedzhidova A.G., Atanadze S.N., Semiletov Y.A., Shkurko T.V., Burkov A.N., Ulanova T.I., Pimenov V.K., Novikov V.V., Khudyakov Y.E., Fields H.A., Kushch A.A. 2002. Characterization of monoclonal antibodies and epitope mapping of the NS4 protein of hepatitis C virus. Immunol. Lett. 83, 187–196.CrossRefPubMedGoogle Scholar
  15. 15.
    Enshell-Seijffers D., Smelyanski L., Gershoni J.M. 2001. The rational design of a “type 88” genetically stable peptide display vector in the filamentous bacteriophage fd. Nucleic Acids Res. 29, 50–63.CrossRefGoogle Scholar
  16. 16.
    Masalova O.V., Abdulmedzhidova A.G., Atanadze S.N., Lakina E.I., Semiletov Yu.A., Burkov A.N., Ulanova T.I., Novikov V.V., Pimenov V.K., Filds G., Khudyakov Yu.E., Kushch A.A. 2002. Characterization of a panel of monoclonal antibodies and the epitope mapping of hepatitis C virus proteins. Dokl. Akad. Nauk. 383, 545–550.Google Scholar
  17. 17.
    Enshell-Seijffers D., Denisov D., Groisman B., Smelyanski L., Meyuhas R., Gross G., Denisova G., Gershoni J.M. 2003. The mapping and reconstitution of a conformational discontinuous B-cell epitope of HIV-1. J. Mol. Biol. 334, 87–101.CrossRefPubMedGoogle Scholar
  18. 18.
    Pereboeva L.A., Pereboev A.V., Wang L.F., Morris G.E. 2000. Hepatitis C epitopes from phage-displayed cDNA libraries and improved diagnosis with a chimeric antigen. J. Med. Virol. 60, 144–151.CrossRefPubMedGoogle Scholar
  19. 19.
    Ivanisenko V.A., Eroshkin A.M. 1997. Search for sites containing functionally significant substitutions in the sets of related or mutant proteins. Mol. Biol. 31, 880–887.Google Scholar
  20. 20.
    Ching W.-M., Wychowski C., Beach M.J., Wang H., Davies C.L., Carl M., Bradley D.W., Alter H.J., Feinstone S.M., Shin J.W.-K. 1992. Interaction of immune sera with synthetic peptides corresponding to the structural protein region of hepatitis C virus. Proc. Natl. Acad. Sci. USA. 89, 3190–3194.PubMedGoogle Scholar
  21. 21.
    Masalova O.V., Atanadze S.N., Kalinina T., Petrakova N.V., Smirnov V.D., Fields G., Khudyakov Yu.E., Kushch A.A. 1996. Immunochemical properties and specificity of monoclonal antibodies with respect to the N-and C-terminal regions of the recombinant hepatitis C virus nucleocapsid protein. Vopr. Virusol. 4, 150–156.Google Scholar
  22. 22.
    Masalova O.V., Kushch A.A. 2003. Monoclonal antibodies to hepatitis C proteins: A tool for mapping antigenic determinants, diagnosing hepatitis C, and studying viral pathogenesis. Ross. Bioterapevt. Zh. 3, 7–23.Google Scholar
  23. 23.
    Cerino A., Boender P., La Monica N., Rosa C., Habets W., Mondelli M.U. 1993. A human monoclonal antibody specific for the N terminus of the hepatitis C virus nucleocapsid protein. J. Immunol. 151, 7005–7015.PubMedGoogle Scholar
  24. 24.
    Siemoneit K., Cardoso M.S., Wolp A., Koerner K., Subanek B. 1994. Isolation and epitope characterization of human monoclonal antibodies to hepatitis C virus core antigen. Hybridoma. 13, 9–13.PubMedGoogle Scholar
  25. 25.
    Menez R., Bossus M., Muller B.H., Sibai G., Dalbon P., Ducancel F., Jolivet-Reynaud C., Stura E.A. 2003. Crystal structure of a hydrophobic immunodominant antigenic site on hepatitis C virus core protein complexed to monoclonal antibody 19D9D6. J. Immunol. 170, 1917–1924.PubMedGoogle Scholar
  26. 26.
    Mondelli M.U., Cerino A., Boender P., Oudshoorn P., Middeldorp J., Fipaldini C., La Monica N., Habets W. 1994. Significance of the immune response to a major, conformational B-cell epitope on the hepatitis C virus NS3 region defined by a human monoclonal antibody. J. Virol. 68, 4829–4836.PubMedGoogle Scholar
  27. 27.
    Chen M., Sallberg M., Sonnerborg A., Jin L., Birkett A., Peterson D., Weiland O., Milich D.R. 1998. Human and murine antibody recognition is focused on the ATPase/helicase, but not the protease domain of the hepatitis C virus nonstructural 3 protein. Hepatology. 28, 219–224.PubMedGoogle Scholar
  28. 28.
    Simmonds P., Rose K.A., Graham S., Chan S.-W., McOmish F., Dow B.C., Follett E.A.C., Yap P.L., Marsden H. 1993. Mapping of serotype-specific, immunodominant epitopes in the NS-4 region of hepatitis C virus (HCV): Use of type-specific peptides to serologically differentiate infections with HCV types 1, 2, and 3. J. Clin. Microbiol. 31, 1493–1503.PubMedGoogle Scholar
  29. 29.
    Chang J.C., Seidel C., Ofenloch B., Jue D.L., Fields H.A., Khudyakov Y.E. 1999. Antigenic heterogeneity of the hepatitis C virus NS4 protein as modeled with synthetic peptides. Virology. 257, 177–190.CrossRefPubMedGoogle Scholar
  30. 30.
    Garnier J., Osguthorpe D.J., Robson B. 1978. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J. Mol. Biol. 120, 97–120.CrossRefPubMedGoogle Scholar
  31. 31.
    Dou X.-G., Talekar G., Chang J., Dai X., Li L., Bonafonte M.-T., Holloway B., Fields H.A., Khudyakov Y.E. 2002. Antigenic heterogeneity of the hepatitis C virus NS5A protein. J. Clin. Microbiol. 40, 61–67.CrossRefPubMedGoogle Scholar
  32. 32.
    Hopp T.P., Woods K.R. 1981. Prediction of protein antigenic determinants from amino acid sequences. Proc. Natl. Acad. Sci. USA. 78, 3824–3828.PubMedGoogle Scholar
  33. 33.
    Enomoto N., Sakuma I., Asahina Y., Kurosaki M., Murakami T., Yamamoto C., Izumi N., Marumo F., Sato C. 1995. Comparison of full-length sequences of interferon-sensitive and resistant hepatitis C virus Ib sensitivity to interferon is conferred by amino acid substitutions in the NS5A region. J. Clin. Invest. 96, 224–230.PubMedGoogle Scholar
  34. 34.
    Bhattacherjee Y., Prescott L.F., Pike I., Rodgers B., Bell H., El-Zayadi A.R., Kew M.C., Conradie J., Lin C.K., Marsden H., Saeed A.A., Parker D., Yap P.L., Simmonds P. 1995. Use of NS4 peptides to identify type-specific antibody to hepatitis C virus genotypes 1, 2, 3, 4, 5 and 6. J. Gen. Virol. 76, 1737–1748.PubMedGoogle Scholar
  35. 35.
    Yip Y.L., Smith G., Ward R.L. 2001. Comparison of phage pIII, pVIII and GST as carrier proteins for peptide immunization in BALB/c mice. Immunol. Lett. 79, 197–202.CrossRefPubMedGoogle Scholar
  36. 36.
    Meola A., Delmastro P., Monaci P., Luzzago A., Nicosia A., Felici F., Cortese R., Galfrl G. 1995. Derivation of vaccines from mimotopes: Immunologic properties of HBsAg mimotopes displayed on filamentous phage. J. Immunol. 154, 3162–3172.PubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2006

Authors and Affiliations

  • E. A. Rechkina
    • 1
  • G. F. Denisova
    • 2
  • O. V. Masalova
    • 1
  • L. F. Lideman
    • 1
  • D. A. Denisov
    • 2
  • E. I. Lesnova
    • 1
  • R. I. Ataullakhanov
    • 3
  • S. V. Gurianova
    • 4
  • A. A. Kushch
    • 1
  1. 1.Ivanovsky Institute of VirologyRussian Academy of Medical SciencesMoscowRussia
  2. 2.Department of Cell Research and Immunology, Wise Faculty of Life SciencesTel-Aviv UniversityTel-AvivIsrael
  3. 3.Immapharma Joint-Stock CompanyMoscowRussia
  4. 4.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia

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